TWI249158B - Trackable optical discs with concurrently readable analyte material - Google Patents

Abstract

Design, manufacture and use of optical discs that permit the concurrent and discriminable acquisition of signals from both operational structures and nonoperational structures is presented. The disc geometries and tracking schemes permit such discs to be read in, and data encoded by nonoperational structures reported by, standard (or minimally-modified) optical disc readers. Single data layer first and second surface discs are described, as are multiple data layer discs. Use of the discs in analyte-specific assay is presented.

Description

1249158 九、發明說明： 【發明所屬之技術領域】 本發明係關於光碟設計和製造以及在光碟閱讀機和讀寫 器之使用。特定地是，本發明係關於允許由光碟非操作結 構如特定分析物驗定信號元素以及由光碟可操作結構如尋 執結構同時及可鑑別擷取信號的光碟設計、製造和使用。 【先前技術】 在過去十年間，掃描共焦雷射顯微術（SCLM)已經將生 命科學成像加以改革。在掃描共焦雷射顯微術中，雷射光 在準確選擇之焦平面上橫跨樣品掃描。由樣品反上的雷射 光被收集，藉由排除所選擇之焦平面以外反射的雷射光、 閃光被移除，而由全厚度、非固定組織和細胞產生清晰的 區段影像。除此以外，電腦驅動之掃描處理之重製空間精 密度允許單獨擷取之區段影像的正確空間登記，藉由各自 擷取區段影像之重疊而允許三維影像之重建。 Wiesendanger, Scanning Probe Microscopy and Spectroscopy: Methods and Applications，劍橋大學出版（1995年 7月）； Cullander，J. Investig. Dermatol. Symp. Proc. 3:166 -171(1998) ； Paddock, Proc. Soc. Exp. Biol. Med. 213:24-31(1996); Ockleford，J· Pathol· 176:1-2(1995); Laurent等 人，Biol. Cell. 80:229-240(1994)。 雷射激發螢光染料和蛋白質之使用為特定配基探針已經 允許掃描雷射顯微術調適為超越標準細胞和組織成像成為 廣泛變化之驗定。因此，雷射掃描細胞計量器已經證明特 98815.doc 1249158 別有利於以螢光為基準之細胞計量驗定細胞循環事件。 Juan 等人，Methods Mol Biol·，91:67-75(1996) ; Juan 等 人，Cell Biol. 2:261-273(1998) ; Juan 等人，Cell Biol. 2:341-350(1998) ; Catch等人，Cytometry 34:36-38(1998); Luther 等人，Microscopy & Micro analysis 3: 235- 236(1997) 〇1249158 IX. DESCRIPTION OF THE INVENTION: TECHNICAL FIELD OF THE INVENTION The present invention relates to the design and manufacture of optical discs and their use in optical disc readers and readers. In particular, the present invention relates to the design, manufacture, and use of optical discs that permit simultaneous verification of signal elements by optical disc non-operational structures, such as specific analytes, and simultaneous and identifiable capture signals from optical disc operable structures, such as ergonomic structures. [Prior Art] Over the past decade, scanning confocal laser microscopy (SCLM) has revolutionized life science imaging. In scanning confocal laser microscopy, laser light is scanned across the sample in an accurately selected focal plane. The laser light from the opposite side of the sample is collected, and by removing the laser light reflected outside the selected focal plane, the flash is removed, and a clear segment image is produced from the full thickness, non-fixed tissue and cells. In addition, the reworked spatial precision of the computer-driven scanning process allows for the correct spatial registration of the separately captured segment images, allowing for the reconstruction of the three-dimensional images by overlapping the segment images. Wiesendanger, Scanning Probe Microscopy and Spectroscopy: Methods and Applications, Cambridge University Press (July 1995); Cullander, J. Investig. Dermatol. Symp. Proc. 3:166-171 (1998); Paddock, Proc. Soc. Exp Biol. Med. 213: 24-31 (1996); Ockleford, J. Pathol. 176: 1-2 (1995); Laurent et al, Biol. Cell. 80: 229-240 (1994). The use of laser-excited fluorescent dyes and proteins for specific ligand probes has allowed scanning laser microscopy to be adapted to standard cell and tissue imaging to become widely variable assays. Therefore, laser scanning cell meters have proven to be particularly useful for fluorescence-based cytometry to determine cell cycle events. Juan et al, Methods Mol Biol, 91: 67-75 (1996); Juan et al, Cell Biol. 2: 261-273 (1998); Juan et al, Cell Biol. 2: 341-350 (1998); Catch et al, Cytometry 34: 36-38 (1998); Luther et al, Microscopy & Micro analysis 3: 235-236 (1997) 〇

Ashby等人的美國專利案號為第5,549,588號專利說明掃 描雷射顯微驗定之’’基因組報告矩陣”。在這些基因組報告 矩陣中，可空間定址矩陣之各元素包含細胞，其中共同螢 光報告之陳式由不同的翻譯調節元素驅動。在掃描期間擷 取的螢光信號強度辨識由各個可空間辨識之翻譯調節元素 驅動的基因位準。 掃描雷射顯微術亦已經調適為建立在矽晶片上的核酸微 陣列之掃描，Lashkari等人，Proc. Natl. Acad. Sci. USA 94:13057-62(1997) ; DeRisi 等人，Science，278:680-86(1997) ; Wodicka等人，Nature Biotechnology，15:1359-67(1997);調適為在細胞中離子通量之量測，Schild，Cell. Calcium 19:281-296(1996) ; Turner 等人，J. Investig. Dermatol. Symp. Proc· 3:136-142(1998);以及調適為各種 細胞成分之次細胞分佈的量測，Takubo等人， Haematologica 82:643-647(1997) ° 這些SCLM應用中的各應用需要電腦控制光學設備的特 殊件。因此在相關技藝中需要存在便宜的一般裝置，該裝 置允許微觀樣本之電腦驅動共焦雷射掃描。 98815.doc 1249158 此種裝置之最小機械需求-雷射、聚焦和偵測光學、精 密度掃描裝置和電腦介面-全部可以在標準光碟閱讀機或 是撰寫器中發現。光碟閱讀機和讀寫器，如CD和DVD將 光由固態雷射聚焦在旋轉光碟表面上以及掃描該光碟以偵 測在可空間定址圖樣次微米結構中數位式編碼之資訊。 光碟和光碟讀寫器調適為雷射掃描圍觀之應用將呈現已 存在方式之上的顯著優點。在這些優點之間的主要優點為 廣泛之應用性和低成本。目前全世界CD和DVD-ROM閱讀 機安裝基座預估為大約3億單元，期望在未來5年將成長為 超過 5億單元。Optical Publishing Industry Assessment，第 9版（Infotech，Inc·, Woodstock，Vermont)(1998)。該裝置為 遍存、便宜和可靠。 使用光碟於微觀結構之偵測和特徵化的其他優點在W0 96/09548(Gordon)、EP A 392475(Idemitsu)、EP A 417 305(Idemitsu) 、 EP A 504432(Idemitsu) 、 WO 98/28623(Gamera)和 WO 98/12559(Demers)中討論，所有這 些文獻在此處以提及的方式併入本文中。其他優點在共同 擁有以及尚在審理中的美國專利申請案案號為第 08/888,935號，1997年 7月 7日申請；第 09/064,636 號，1998 年4月21曰申請；第09/120,049號，1998年7月21曰申請； 以及發佈為 WO 98/3 85 10，WO 98/385 10和 WO 98/01533之 對等國際專利；這些文獻在此處以提及的方式併入本文 中〇 然而，雖然光碟閱讀機擁有有效共焦雷射顯微掃描之機 98815.doc 1249158 械式先决條件，但現存光碟閱讀機之操作需求呈現在光碟 表面上配置之微觀結構成功债測和特徵化的重大障礙/、 至；具有4個操作區需求必須滿足於光碟以正確地讀取 和解碼呈現在光碟内的資料：該閱讀機必須正確地聚隹在 將資料編碼的光碟平面上、必須控制光學拾音器的徑向位 置、必須控制光學拾音器的切線位置以及必須控制光碟的 轉數。最普通的光碟系統使用光學媒體本身滿足這些需求 中的至少某些需求。 因此在典型壓製的⑶中，光碟機板以由-系列浮凸1 洞組成之螺旋磁軌壓印，由此產生的信號為光碟閲讀機利 用:維持適當的焦點和尋軌。在CD_R中，由使用者寫入 的貝枓編碼染料標記提供在依序讀取時的必要尋軌結構。 =-般地是’在已存在光碟標準之各標準中，用以編碼資 1 之結構同時作用為提供該閱讀機控制其操作所需之操作 2號。雖然有效率，但是此種標準沒有提供用於由配置在 一碟上之非操作結構擷取資料的規定。 例如’因為尋軌結構強制嵌入在光碟資訊層 用於光碟之雷射畀拉、^ φ η λ應 雷射最接近表面的結構可能㈣此種操作 之谓測，以及因此妨礙閱讀機的正確操作。甚至 焉 :操作結構可能置於該光碟操作結構焦平面之足夠外，而 止由该閱讀機光學拾音器對其同時和可鑑別之_。 :問題的一個解決方案為使非標準傳動裝置。 傳動裝置利用二光學拾音 ° 另-個目“ 尤子^曰盗，其令之一谓測尋軌資訊， 補測表面結構’ EP A 392475(Idemitsu)、Ερ Α 9«815.doc 1249158 417 305(Idemitsu)、EP A 504432(Idemitsu)和 EP A 417 3〇5(Idemitsu)。然而，此種修正假設使用光碟閱讀機於雷 射Μ觀谓測之主要優點，為此種裝置的普遍分伟。 ，光碟相關技藝中存在需要允許使標準光碟閱讀機/讀 寫為由光碟非操作結構如配置在光碟上的特定分析物信號 疋素同時和任意顧取信號，而信號由光碟操作結構如 結構產生。 【發明内容】 本^明藉由提供光碟、製造該光碟方法以及 :法而解決相關技藝這些和其他問題，其t該光碟： =閱讀機/讀寫器由光碟非操作結構如配置在光碟上的特 物信號元素同時和可鑑別地搁取信號，而信號由光 碟#作結構如尋執結構產生。 =^刀依據非操作結構能夠和光碟操作結構共焦配 解^以及雷射最接近於先前應用之反射層，因此 幾、^、維持焦點在操作和非操作結構上的光學問題。此 製造Λ光碟而達成，其中該光碟完整基板之圖像 為：杌(也:尤疋:兒’促成光碟之資訊層之該基板表面)呈現 面^ 4 # |面（”倒置光碟°因為在操作焦平 ^貫貝上小於在光碟第一表面上的光束半 &二倒置光碟以新的 成為可能。 <使^米大小之非操作使用 產生呈右^夠猎由利用傳統母片為壓模加以製造，因此 正浮雕呈現之操作結構的前向影像倒置光碟。 98815.doc 1249158 第一製造處理，其中利用為壓模之父片或是子片亦能夠加 、利用在此苐一製造處理中，主片具有與傳統相反之影 像，泫形成之前向影像倒置光碟具有以負浮雕呈現的操作 結構。The U.S. Patent No. 5,549,588 to Ashby et al. describes the scanning of the laser microscopic verification of the 'genomic reporting matrix.' In these genome reporting matrices, the elements of the spatially addressable matrix contain cells, with a common fluorescent report. The pattern is driven by different translational adjustment elements. The intensity of the fluorescent signal captured during the scan is identified by the genetic level driven by each spatially identifiable translational regulatory element. Scanning laser microscopy has also been adapted to Scanning of nucleic acid microarrays on wafers, Lashkari et al, Proc. Natl. Acad. Sci. USA 94: 13057-62 (1997); DeRisi et al, Science, 278: 680-86 (1997); Wodicka et al, Nature Biotechnology, 15: 1359-67 (1997); Adaptation to measurement of ion flux in cells, Schild, Cell. Calcium 19:281-296 (1996); Turner et al., J. Investig. Dermatol. Symp. Proc. 3:136-142 (1998); and measurement of subcellular distribution adapted to various cellular components, Takubo et al, Haematologica 82:643-647 (1997) ° Computer-controlled optics for applications in these SCLM applications Special parts of the equipment There is therefore a need in the art for an inexpensive general device that allows computer-driven confocal laser scanning of microscopic samples. 98815.doc 1249158 Minimum mechanical requirements for such devices - laser, focus and detection optics, precision scanning Devices and computer interfaces - all found in standard CD readers or writers. Optical disc readers and readers, such as CDs and DVDs, focus light from a solid-state laser onto the surface of a rotating disc and scan the disc for detection Information on digital encoding in spatially-addressable sub-micron structures. The application of optical disc and optical disc readers to laser scanning on-the-go applications will present significant advantages over existing methods. The main advantages between these advantages are extensive Applicability and low cost. Currently, the world's CD and DVD-ROM reader installation base is estimated to be about 300 million units, and it is expected to grow to more than 500 million units in the next five years. Optical Publishing Industry Assessment, 9th Edition ( Infotech, Inc., Woodstock, Vermont (1998). The device is ubiquitous, inexpensive and reliable. Use of optical discs for microstructure detection and Other advantages of the characterization are in WO 96/09548 (Gordon), EP A 392475 (Idemitsu), EP A 417 305 (Idemitsu), EP A 504432 (Idemitsu), WO 98/28623 (Gamera) and WO 98/12559 (Demers). In the discussion, all of these documents are incorporated herein by reference. Other advantages are co-owned and pending US Patent Application No. 08/888,935, filed July 7, 1997; No. 09/064,636, filed April 21, 1998; 09/120,049 No., filed July 21, 1998; and equivalent international patents issued as WO 98/3 85 10, WO 98/385 10 and WO 98/01533; these documents are hereby incorporated herein by reference. However, although the optical disc reader has an effective confocal laser microscopy machine 98815.doc 1249158 mechanical prerequisites, the operational requirements of the existing optical disc reader appear on the surface of the optical disc to configure the microstructure for successful debt testing and characterization. Obstacle/, to; having 4 operating area requirements must be satisfied with the disc to correctly read and decode the data presented on the disc: the reader must be properly clustered on the disc surface where the data is encoded, the optical pickup must be controlled The radial position, the tangential position of the optical pickup must be controlled, and the number of revolutions of the disc must be controlled. The most common optical disc systems use optical media to meet at least some of these needs. Therefore, in the typical press (3), the optical disk board is embossed with a spiral track composed of a series of embossed holes of one hole, and the resulting signal is used for the optical disk reader: maintaining proper focus and tracking. In CD_R, the beggar coded dye mark written by the user provides the necessary tracking structure for sequential reading. = - Generally, in the various standards of the existing optical disc standard, the structure for encoding the resource 1 simultaneously functions to provide the operation No. 2 required for the reader to control its operation. Although efficient, such standards do not provide provisions for extracting data from non-operational structures configured on a disc. For example, 'because the tracking structure is forced to be embedded in the disc information layer for the laser pull of the optical disc, ^ φ η λ should be the structure of the laser closest to the surface. (4) The operation of this operation, and thus hinder the correct operation of the reader . Even 焉: the operating structure may be placed outside of the focal plane of the optical disc operating structure, while being simultaneously and identifiable by the optical pickup of the reader. : A solution to the problem is to make non-standard transmissions. The transmission uses two optical pickups. Another one is "Ultra 曰 曰 , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , 305 (Idemitsu), EP A 504432 (Idemitsu) and EP A 417 3〇5 (Idemitsu). However, this modification assumes that the main advantage of using a disc reader in laser viewing is that it is a common part of such a device. Wei. There is a need in the optical disc related art to allow a standard optical disc reader/reading to be a non-operating structure of a disc, such as a specific analyte signal, which is disposed on the optical disc, simultaneously and randomly take signals, and the signal is operated by a disc operating structure such as The present invention provides a method for solving the related art and other problems by providing a compact disc, a method for manufacturing the optical disc, and a method, and the optical disc: = the reader/writer is configured by a non-operating structure of the optical disc, such as a configuration The special signal element on the optical disc simultaneously and identifiably captures the signal, and the signal is generated by the optical disc # as a structure such as a search structure. The ^^ knife can be combined with the optical disc operating structure and the laser according to the non-operating structure. most Close to the reflective layer of the previous application, therefore, the optical problem of maintaining the focus on the operational and non-operational structures. This is achieved by manufacturing a compact disc, wherein the image of the complete substrate of the optical disc is: 也 (also: 疋: The surface of the substrate that promotes the information layer of the disc is presented ^ 4 # | face ("inverted disc ° because the beam is smaller than the beam half & on the first surface of the disc in the operation of the coke It is possible to make the non-operational use of the size of the meter to produce a right-handed image that is manufactured by using a conventional master piece as a stamper, so that the forward-looking image of the operational structure of the relief is presented. 98815.doc 1249158 A manufacturing process in which a parent or a sub-piece that is a stamper can be added and utilized in the manufacturing process, the main film has an image opposite to the conventional one, and the image is inverted to the image before the formation of the image has a negative relief Operational structure.

因此，在第一特徵中，本發明呈現包括基板之光碟，該 基板具有@樣化表面和額外表面，該基板圖樣化表面促成 光碟資訊層，纟中當基板圖樣化表面呈現為雷射最接近盆 額外表面時，該資訊層包含可由光碟機尋執之操作表面f 如本發明第—處理製造時，此種倒置光碟之操作結構可 尋軌地配置為正浮雕之前向影像。在此方位中，該光碟之 操作結構可尋執地配置在資訊層平面中，該資tfl層在基板 額外表面最遠端。 *如本發明第二處理製造時，此種倒置光叙操作結構可 :軌地配置為負浮雕之前向影像。在此方位中，該光碟之 才呆作結構可尋執地配置在資 基板額外表面。纟貝'層千面中’该資訊層最接近 在特定具體實施财，該倒置光碟尚包括至少_非操 =構，該結構為和操作結構（特収，尋執）㈣取。 體實施例之特定具體實施财，該非操作結構可 2特疋“斤物信號元素，和光碟操作結構共焦配置。 在另—特徵中，本發明提供製造此種倒置光 法。因此，本發明提供用於製造 心裏圮方 光碟的方法，該光碟基板具有：：，之倒置 括:形成具有可尋執操作結構之壓二:和:::: 988l5.doc -11- 1249158 具有相I影像方位及浮雕；w及之後直接由該壓模形成光 碟基板。在第二方法中，本發明提供製造前向影像、負浮 雕之倒置光碟的方法，該光碟基板具有可尋軌操作結構， °亥方法包括.幵> 成相反主片；形成具有和該相反主片相反 影像方位及浮雕之壓模；以及之後直接由該壓模形成光碟 基板。 在另一具體實施例中，前述的新處理的其中之一能夠利 • 用CD-R或是CD-RW主片加以執行（或是與此相反），而鑄模 為DVD鑄模。此種新形成之光碟和⑶鑄模中鑄造的光碟 比較為具有較減少之深度。 本發明尚提供用於製造倒置光碟之方法，該光碟利用倒 置光碟基板而適合於非操作結構之可尋軌讀取。在此種第 一具體實施例中，本發明則共用於製造具有同時可讀取非 操作結構之可尋執光碟，該方法包括：應用反射層至該倒 置光碟基板的圖樣化表面，以及配置和該反射層共焦及雷 _ ㈣近之非操作結構。該倒置光碟基板可以藉由任何前述 加以準備。 對本發明前向影像/正浮雕倒置光碟或是前向影像/負浮 雕倒置光碟而言，沒有透明基板層為雷射接近至該光碟資 訊層。所以額外裝置為所需以改變光學路徑之長度，因此 加強光碟反射層表面上入射雷射之聚焦。此方式可以藉由 附接非整體或整體雷射折射構件為倒置光碟雷射接近側之 蓋板而完成以建立光碟組件。該蓋板作用為折射，以及因 此更進一步將入射光束聚焦在光碟操作平面上。 98815.doc -12 - 1249158 因此，在另一特徵中，本發明提供具有可讀取非操作結 構之光碟組件，該組件包括··倒置光碟和蓋板，其中該蓋 板更進一步將入射光束聚焦在光碟資訊層上。在一些具體 貫施例中，該蓋板為非整合至該光碟以及可由此附接。在 其他具體實施例中，該蓋板為整合至該光碟。在一些具體 實施例中，該蓋板本質上由塑膠組成、本質上由破璃組 成、或是本質上由矽組成。 本發明提供達成操和作非操作態樣共焦配置之其他方 因此，在另-特徵中，光碟操作結構之影像，特別是可 尋執結構之影像為全像投影至與非操作結構共焦之平面 ^在較佳具體實施例中，使用反射全像。全像之優點包 3光碟上操作結構和麵作結構之實體分離。 此種實體分離之額外方式為利用多層DVD技術。Accordingly, in a first feature, the present invention presents a disc comprising a substrate having a @formed surface and an additional surface, the substrate patterned surface contributing to the disc information layer, wherein the substrate patterned surface appears to be the laser closest When the extra surface is filled, the information layer includes an operation surface f which can be searched by the optical disc. When the manufacturing process is performed as in the first process of the present invention, the operation structure of the inverted optical disc can be configured to be a front relief image. In this orientation, the operating structure of the optical disc is configurably disposed in the plane of the information layer, the tfl layer being at the farthest end of the additional surface of the substrate. * When the second process of the present invention is manufactured, such an inverted optical operation structure can be configured to be a negative relief frontward image. In this orientation, the disc is constructed to be configurably configured on the additional surface of the substrate. The mussel's layer is the closest to the information layer. In the specific implementation, the inverted disc still includes at least _ non-operational structure, and the structure is taken with the operation structure (special collection, search) (four). For a specific implementation of the embodiment, the non-operational structure can be characterized as a "pump signal element, and a disc operating structure confocal configuration. In another feature, the present invention provides for the fabrication of such an inverted photo method. Thus, the present invention Providing a method for manufacturing a heart-shaped optical disc having::, inverted comprises: forming a pressure two with a searchable operation structure: and :::: 988l5.doc -11- 1249158 having a phase I image orientation And embossing; w and then directly forming the optical disc substrate by the stamper. In the second method, the present invention provides a method for manufacturing a front-facing image and a negative embossed inverted optical disc, the optical disc substrate having a track-searching operation structure, Including: 幵> into an opposite main sheet; forming a stamp having an image orientation opposite to the opposite main sheet and embossing; and then directly forming the disc substrate from the stamper. In another embodiment, the aforementioned new processing One of them can be implemented with a CD-R or a CD-RW master (or vice versa), and the mold is a DVD mold. The newly formed disc is compared with the (3) cast disc. There is a reduced depth. The present invention also provides a method for fabricating an inverted optical disk that utilizes an inverted optical disk substrate for tracking search of a non-operational structure. In this first embodiment, the present invention Commonly used to fabricate a rewritable optical disc having a simultaneously readable non-operational structure, the method comprising: applying a reflective layer to a patterned surface of the inverted optical disc substrate, and configuring the reflective layer to have a confocal and lightning _ (four) near non-operational The inverted optical disc substrate can be prepared by any of the foregoing. For the forward image/positive embossed inverted optical disc or the forward image/negative embossed inverted optical disc of the present invention, there is no transparent substrate layer for the laser to approach the optical disc information layer. Therefore, an additional device is required to change the length of the optical path, thus enhancing the focus of the incident laser on the surface of the reflective layer of the optical disc. This can be achieved by attaching a non-integral or integral laser refractive member to the side of the inverted optical disk. The cover is completed to create a disc assembly. The cover acts as a refraction, and thus further focuses the incident beam on the disc operation 98815.doc -12 - 1249158 Accordingly, in another feature, the present invention provides a disc assembly having a readable non-operating structure, the assembly comprising: an inverted disc and a cover, wherein the cover further The incident beam is focused on the disc information layer. In some embodiments, the cover is non-integrated to the disc and can be attached thereto. In other embodiments, the cover is integrated into the disc. In a specific embodiment, the cover is essentially composed of plastic, consists essentially of broken glass, or consists essentially of ruthenium. The present invention provides for other aspects of achieving a non-operating mode confocal configuration, and thus In the feature, the image of the optical disc operating structure, in particular the image of the searchable structure, is a holographic projection to a plane that is confocal with the non-operating structure. In a preferred embodiment, a full reflection image is used. The advantage of the full image package is the physical separation of the operating structure and the surface structure on the optical disc. An additional way of separating such entities is to utilize multi-layer DVD technology.

^ ’在另—特徵中，本發明提供具有可讀取非 ^ 光碟，包括反射層；額外反射層；以及非操作結構， 結槿f反射層或〜亥額外反射層具有光碟閱讀機可尋軌之 以及其中4非㈣結構和可尋軌結構為可讀取地配 置。在與DVD標準相容的典 中之-為半反射，以及兮打;; 層其 射層為共焦地配置。 轉結構和該反㈣或是半反 雖然上述之新方式解決π 建立聚焦的光學問題’ h們太構和非操作結構上 時偵測。第一，這些方式不需要解要允㈣結構之同 个而要解決在呈現此種共焦配置 988I5.doc -13- 1249158 非操作結構時維持尋軌、聚焦和同步（速度控制）之問題。 第二，和操作態樣（例如訊洞）共焦配置之非操作結構可能 4成^號不足以由操作信號區別而防止由此鐘別。 本發明因此更進一步依據在讀取期間產生之電氣信號具 有較先前瞭解地更具有較大資訊内容之新發現。也就 说，邊電氣信號被設計為提供二特定類別資訊_該信號為 光碟操作所需以及該資訊為數位式編碼在該光碟上_事實 φ Jl可以用多種方法查詢和量測以顯露有關該光碟實體結構 先前未開發之豐富資訊。 此外，本發明實際說明這些電氣信號的所需操作成分能 ，確定地分離，因此釋放該信號其餘成分以用於該非操作匕 量測而不干擾成功的驅動操作。 所需之部分信號的分離， 特別是光碟尋執所需之部分信號^ 'In another feature, the present invention provides a readable CD, including a reflective layer; an additional reflective layer; and a non-operating structure, a f-reflecting layer or an additional reflective layer having a disc reader for tracking And wherein the 4 non-four structure and the trackable structure are readable. In the classics compatible with the DVD standard - semi-reflective, and beating;; the layer of the layer is confocal. The rotation structure and the inverse (four) or semi-reverse, although the above-mentioned new way to solve the optical problem of π establishing focus, is detected when the structure is too structured and non-operating. First, these methods do not require the same (4) structure to solve the problem of maintaining tracking, focusing, and synchronization (speed control) when presenting such a confocal configuration 988I5.doc -13-1249158 non-operational structure. Second, the non-operational structure of the confocal configuration with the operational aspect (e.g., the hole) may not be sufficient to be distinguished by the operational signal to prevent the clock from being clocked. The present invention therefore further relies on the discovery that electrical signals generated during reading have new findings that are more informative than previously known. That is to say, the edge electrical signal is designed to provide two specific categories of information _ the signal is required for the operation of the optical disc and the information is digitally encoded on the optical disc _ fact φ Jl can be queried and measured in various ways to reveal the relevant A wealth of information previously undeveloped on the physical structure of the disc. Moreover, the present invention actually illustrates the required operational component of these electrical signals to be surely separated, thereby releasing the remaining components of the signal for the non-operational measurement without disturbing successful drive operations. The separation of some of the required signals, especially the part of the signal required for the optical disc search

該倒置光碟上之非操作結構的有目 因此’在另-特徵中，本發明提供設計為允許光碟操作 些後期光碟幾何允許在 目的、同時和可鑑別偵 988I5.doc -14- 1249158 測。 入本發明因此提供如上述之倒置光碟，其中該操作結構包 έ擺動、、文道。如δ亥製造處處理，該擺動”紋道”以正浮雕呈 ，為前向影像（處理υ或是以負浮雕呈現為前向f彡像。後期 T向加強非操作結構如特定分析物信號元素之定中心為在 吻取期間的尋執結構之上。和光碟資訊層共焦配置以及雷 射最接近之非操作結構為可同時與擺動紋道伯測以及其信 號可由擺動紋道鐘別。 匕呈現之例子2-3貫際說明本發明單一資料層、前向 影像擺動紋道、正浮雕倒置光碟之成功使用以⑴藉由免疫 分析伯測人體A液中的IgG，⑺以利用最小修正之光碟閱 讀機藉由特定免疫學的反應將光碟表面上捕獲的人體红血 球加以谓測以及特徵化。例子7實際說明核酸基礎驗定調 適為由此說明的偵測原理。 【實施方式】 為了使由此說明之本發明完全被瞭解，所以陳述下列詳 細說明。在該說明中，利用下列術語。 當由此使用時，該術語"徑向"表示在至少一光碟資料編 7表面（亦即光碟資訊層光學可偵測表面）的平面中，沿著 尋轨螺旋前進或是後退的方向。如本發明之光碟表面，= 以是内部或是外部表面。 當由此使用時，該術語”切線”表示在至少一光碟資料蝙 碼表面的平面+，沿著由該光碟中心向外周騎的線之向 内或是向外的方向。 ^ 98815.doc -15- 1249158 當由此使用時，該術語’’徑向平面π視為該光碟可尋執（亦 即螺旋尋軌）結構所配置的平面，以及是至少一光碟資料 編碼表面的平面。 當由此使用時，該術語”非操作結構”意義為配置在光碟 表面上或是光碟内的任何結構，當該光碟由光碟閱讀機讀 取時，該結構能夠產生信號，然而該結構信號不是讀取期 間的驅動操作所需（雖然可能為有利）。非操作結構包含， 例如特定分析物信號元素，如下文緊接著所說明。 當由此使用時，該術語π特定分析物信號元素”視為可以 用以發出信號為在應用至光碟之樣本中特定分析物存在的 任何非操作結構。該術語因此尤其包含，由此舉例證明之 該信號元素-包含串珠及細胞-以及在尚審理中及共同擁有 之美國專利申請案案號為08/888,935號，1997年7月7曰申 請；09/120,049號，1998 年 7 月 21 日申請；〇9/183,842號， 1998年10月30日申請；09/311，329，1999年5月14日申請； 60/134,368，1999年 5 月 14曰申請；Siddiqi 等人之"Methods and Apparatus for Physically Patterning Nonoperational Structures on an Optical Disc，n美國臨時專利申請案案號為 第60/150,287 ¾¾，1999年8月23日申請（事務所檔案案號 BURST-14P) ; Worthington 等人之’’Methods and Apparatus for Analyzing Operational and Nonoperational Data Acquired from Optical Discs，’’美國申請案序號為第 09/378,878號,，1999年8月23日申請（事務所檔案案號 BURST-15)；以及 Worthington 等人之，，Methods and 98815.doc -16- 1249158The non-operational structure on the inverted optical disc is therefore in the other feature, the present invention provides a design that allows the optical disc to operate at a later optical disc geometry allowing for the purpose, simultaneous and identifiable detection of 988I5.doc -14-1249158. The invention thus provides an inverted optical disc as described above, wherein the operational structure includes a wobble, tangible track. For example, in the processing of δHai, the oscillating "grain" is presented as a forward embossing, which is a forward image (processed υ or negative embossed as a forward f 彡 image. Late T-direction enhances non-operational structures such as specific analyte signals The center of the element is above the search structure during the kiss. The confocal configuration of the disc information layer and the non-operating structure closest to the laser are simultaneously capable of simultaneous detection with the wobble track and its signal can be tracked by the wobble track. Example 2-3 of the present invention illustrates the successful use of the single data layer, the forward image wobble track, and the embossed inverted optical disc of the present invention (1) by immunoanalysing the IgG in the human body A liquid, (7) to utilize the minimum The modified optical disc reader pre-measures and characterizes the human red blood cells captured on the surface of the optical disc by a specific immunological reaction. Example 7 actually illustrates the adaptation of the nucleic acid basic assay to the detection principle thus explained. The invention thus described is fully understood, so the following detailed description is set forth. In this description, the following terms are utilized. When used thereby, the term "radial" At least one of the surface of the optical disk data 7 (ie, the optically detectable surface of the optical disk information layer) is advanced or retracted along the tracking spiral. The surface of the optical disk of the present invention, = internal or external surface When used herein, the term "tangent" means the plane + on the surface of at least one disc data bat, in the direction of the inward or outward direction of the line riding from the center of the disc to the outer circumference. ^ 98815.doc -15- 1249158 When used herein, the term 'radial plane π is taken to mean the plane in which the disc is searchable (ie, the spiral tracking) structure, and the plane on which at least one of the disc data is encoded. As used herein, the term "non-operating structure" means any structure disposed on the surface of a disc or within a disc that is capable of generating a signal when the disc is read by a disc reader, however the structural signal is not read. The driving operation required during the fetching process (although it may be advantageous). The non-operational structure contains, for example, a specific analyte signal element, as explained immediately below. When used thereby, the term π The "analyte signal element" is considered to be any non-operating structure that can be used to signal the presence of a particular analyte in a sample applied to the disc. The term thus includes, inter alia, the signal element - including beading and cells - and in the trial and co-owned US Patent Application No. 08/888,935, July 7, 1997 application; 09/120,049, July 21, 1998; 〇9/183,842, 1998 Application on October 30th; 09/311,329, May 14, 1999; 60/134,368, May 14, 1999; Siddiqi et al."Methods and Apparatus for Physically Patterning Nonoperational Structures on an Optical Disc, n US Provisional Patent Application No. 60/150,287 3⁄43⁄4, application dated August 23, 1999 (office file number BURST-14P); Worthington et al.''Methods and Apparatus for Analyzing Operational and Nonoperational Data Acquired from Optical Discs, ''US application number No. 09/378,878, filed on August 23, 1999 (office file number BURST-15); and W Orthington et al., Methods and 98815.doc -16- 1249158

Apparatus for Optical Disc Data Acquisition Using Physical Synchronization Markers，”美國臨時專利申請案案序號為第 號，1999年8月23曰申請（事務所檔案案號 BURST-18P)的那些信號元素；這些專利申請案中的文獻 在此處以提及的方式併入本文中。該術語包含單獨由光碟 閱讀機可偵測的那些結構以及需要額外組件而成為可偵測 的那些結構。 為提供用於本發明更進一步討論的一些背景，傳統光碟 之突起結構和圖1有關聯地加以簡短說明。 圖1A描繪光學拾音器（接物組件）10和標準CD_類別光碟 11，而在二組件之間的光路徑以虛線指示。為了明確性， 圖1A描繪光學拾音器組件的最小配對件。圖⑺提供光碟 11相對於入射雷射為相同方位的側面橫斷面放大圖。 參考圖1A和圖1B，該光學拾音器包含雷射源19、透鏡 12-14、光束***器15、四分波片2〇和偵測器18。雷射源 1 9典型地為雷射二極體，發射雷射光束而經由光束***器 1 5傳輸以及之後由透鏡12照準。該照準之光束之後由光束 ***器15反射而往前至光碟丨丨。物鏡13將雷射光束聚焦在 光碟11雷射接近或是第一表面上的小點。 雷射光束由光碟反射層114(在傳統CD-類別中，第二表 明）表面反射以及經由物鏡13和四分波片20回轉至光束分 裂器15。光束***器15導引反射之雷射光束經由透鏡14， 該透鏡將聚焦在偵測器1 8上。散光元件16可以被包含在光 束***器15與偵測器18之間以導入在反射之雷射光束中的 98815.doc -17- 1249158 散光。 如圖1B之更詳細顯示，CD-類別光碟11包括3實體層·· 由雷射接近至雷射遠端為透明基板112、反射層U4和防護 層116。各層組合總深度微不足道地為12 mm。該圖為不 成比例加以繪晝。 當沿著光軸量測時，透明基板112組成典型Cd_類別光碟 1 ·2 mm厚度的大部分，以及作用為光學和結構功能。這些 _ 雙重角色4曰疋该基板112具有平面表面和圖樣化表面··而 傳統光碟在傳統讀取期間，平面表面呈現為基板二表面之 雷射接近表面，以及圖樣化表面呈現為基板二表面之雷射 遠端表面。 相對於其光學功能，透明基板112的折射性質作用為將 單色系統中的光學路徑縮短；基本上，透明基板112更進 步作用為將入射雷射聚焦在反射層U4上：在CD-類別光 碟雷射接近表面或是第一表面上，雷射點具有大約8〇〇 μιη •的直徑；透明基板112更進一步將光束聚焦，在此光碟第 二表面之反射表面114上達成大約h7 μηι的直徑。在典型 CD中，透明基板112主要由聚碳酸脂組成，以及因此具有 1 ·55的折射指數。對熟知相關技藝之人士而言，可以使用 除聚碳酸脂以外之其他材料是明顯的，只要材料厚度和折 射“數知:供光碟閱項機聚焦系統足夠的辅助以確定聚焦在 反射層114上。 透明基板112亦提供光碟丨丨的主要結構整合性。反射層 114厚度大約為〇.〇5至〇·ι μηι，以及防護層116典型地包括 98815.doc -18- 1249158 亮漆材料，當外露在uv光線時將變硬，厚度介於1〇與3〇 微米之間。因此，透明基板112組成光碟的主實體層，以 及是唯-能夠傳獸足夠剛性至光碟以保留光碟操作結構的 一層0 基板層112典型以螺旋磁㈣製，該磁軌由光碟最内部 可讀取部分往外螺旋至光碟最外部可讀取部分。在不可記 錄光碟中，此磁軌為由-系列訊洞組成，各訊洞具有大約 為光線波長"4倍的深度，用以讀取該光碟。該訊洞具有 變化長度，該訊洞長度和間隔將資料加以編碼。如下文更 進一步討言余’標_(：1)1可記錄光諜包含螺旋紋道而非訊 洞。此種擺動紋道118的一些部分橫斷面圖以圖⑴的透視 圖加以圖不顯示。 反射層114厚度大約為0·05至〇·!微米，以及典型地包括 反射金屬材料’如銘、銀、金或鋼。反射層ιΐ4為典型地 經由磁電管噴灑處理應用至基板112圖樣化表面。另一種 處理，蒸敷法可加以使用，但是較慢得多。化學濕法，，鍍 、艮處理（使用銀、鎳或是其他金屬）亦可以加以使用以形成 在透明基板112圖樣化表面上的反射層114。 防護層116典型地包括亮漆材料，當外露在紫外線（υν) 光線時，稱為"curing”處理，材料將變硬，厚度介於1〇與 3〇微米之間。防護層116作用為防止反射層ιΐ4刮傷和氧 化以及提供標號可以印在上面的方便表面。防護層丨j 6 典型地經由自旋鍍層處理應用至透明基板112和反射層 114，因此當外露在UV光線時，小量材料喷灑在接近反射 98815.doc -19- 1249158 層Π4内徑的光碟上，而且該光碟以高速旋轉，而使薄層 材料覆蓋在光碟表面上。之後，光碟外露在UV光線，而 使亮漆材料變硬。 雖然包括3實體層，光碟11亦被視為具有”資訊層，，（亦為，， 資料層”）115 ’至少為透明基板112和反射層114實體促成該 層。 n資訊層，，由二平面界定。對由此之目的而言，，，平面A” 為雷射最接近徑向平面而”平面B”為雷射最遠端徑向平 面，該操作結構延伸至該平面。在標準CD或是單一層 OVD中，只具有單一資訊層，以及該光碟因此傳統上描述 為單一資料層（或是，，單一層”）光碟。 各種不同CD和DVD標準預期具有微不足道深度（由光軸 疋義之方向）為1.2 mm以及徑向大小為120 mm的微不足道 直徑光碟。 雖然该微不足道厚度為丨·2 mm，光碟技術的資深標準 (口語化為，”紅皮書”），重新發行為IEC 908，允許厚度為 1 3-1.5 mm(用於所有層組合）。然而，閱讀機能夠容納一 二額外變化，以及適合由CD和DVD驅動裝置讀取之光碟 可以具有最大大約為2.4 mm以最小大約為微不足道〇8 mm 之深度’較佳為1·〇-1·4 mm，更佳地為ΐ·υ·3 mm，最佳 地為1.2 mm。 相對於CD和DVD直徑，該標準設定120 mm的微不足道 直亡。然而，光碟閱讀機可以容納徑向直徑為丨〇〇_丨4〇 的光碟，較佳為丨1(M30 mm，更佳地為丨15_125 mm，最佳 988l5.doc 1249158Apparatus for Optical Disc Data Acquisition Using Physical Synchronization Markers, "The US Provisional Patent Application No. is the number, and the signal elements of the application (office file number BURST-18P) of August 23, 1999; in these patent applications The literature is hereby incorporated by reference in its entirety. The term includes those structures that are detectable by a disc reader alone and those that require additional components to be detectable. Further discussion for providing the present invention Some backgrounds, the protruding structure of the conventional optical disc is briefly described in association with Fig. 1. Fig. 1A depicts an optical pickup (splicing assembly) 10 and a standard CD_class optical disc 11, and the optical path between the two components is indicated by a broken line. For clarity, Figure 1A depicts the minimum counterpart of an optical pickup assembly. Figure (7) provides a side cross-sectional enlarged view of the optical disc 11 in the same orientation relative to the incident laser. Referring to Figures 1A and 1B, the optical pickup includes a laser. Source 19, lens 12-14, beam splitter 15, quarter wave plate 2〇, and detector 18. Laser source 19 is typically a laser The polar body, which emits the laser beam, is transmitted via the beam splitter 15 and then is directed by the lens 12. The collimated beam is then reflected by the beam splitter 15 forward to the disc. The objective lens 13 focuses the laser beam on the disc. 11 The laser is close to or a small spot on the first surface. The laser beam is reflected by the surface of the disc reflection layer 114 (in the conventional CD-class, second indication) and is rotated to the beam splitting via the objective lens 13 and the quarter-wave plate 20 The beam splitter 15 directs the reflected laser beam through the lens 14, which will be focused on the detector 18. The astigmatism element 16 can be included between the beam splitter 15 and the detector 18 for introduction. 98815.doc -17-1249158 astigmatism in the reflected laser beam. As shown in more detail in FIG. 1B, the CD-class optical disc 11 includes a 3 physical layer, which is close to the laser distal end and is a transparent substrate 112. Reflective layer U4 and protective layer 116. The total depth of each layer combination is negligibly 12 mm. The figure is disproportionately drawn. When measured along the optical axis, the transparent substrate 112 constitutes a typical Cd_class disc 1 · 2 mm thickness Most of it, and the role For optical and structural functions. These _ dual roles 4 曰疋 the substrate 112 has a planar surface and a patterned surface · · Traditional optical disc during the conventional reading, the planar surface appears as the laser surface close to the surface of the two surfaces of the substrate, and the patterning The surface appears as a laser distal surface of the two surfaces of the substrate. Relative to its optical function, the refractive nature of the transparent substrate 112 acts to shorten the optical path in the monochromatic system; basically, the transparent substrate 112 is more progressively acting as an incident lightning Focusing on the reflective layer U4: on the CD-class disc laser close to the surface or on the first surface, the laser spot has a diameter of about 8 〇〇μη•; the transparent substrate 112 further focuses the beam, in this disc A diameter of about h7 μηι is achieved on the reflective surface 114 of the two surfaces. In a typical CD, the transparent substrate 112 is mainly composed of polycarbonate, and thus has a refractive index of 1.55. It will be apparent to those skilled in the art that other materials than polycarbonate can be used, as long as the material thickness and refraction are "available for the disc sighting machine focusing system to ensure focus on the reflective layer 114. The transparent substrate 112 also provides the primary structural integrity of the disc. The reflective layer 114 has a thickness of approximately 〇.5 to ι·ι μηι, and the protective layer 116 typically includes 98815.doc -18-1249158 lacquer material. When exposed to uv light, it will harden between 1 〇 and 3 〇 micron. Therefore, the transparent substrate 112 constitutes the main physical layer of the optical disc, and is only capable of transmitting the animal sufficiently rigid to the disc to retain the operating structure of the optical disc. A layer of 0 substrate layer 112 is typically made of spiral magnetic (four), which is spiraled from the innermost readable portion of the optical disc to the outermost readable portion of the optical disc. In a non-recordable optical disc, the magnetic track is composed of a series of channels. Each hole has a depth of about 4 times the wavelength of the light for reading the disc. The hole has a varying length, and the length and interval of the hole encode the data. One step to discuss the remaining 'mark _ (: 1) 1 can record the optical spy including the spiral track instead of the hole. Some parts of the cross-sectional view of the wobble track 118 are not shown in the perspective view of Figure (1). Layer 114 has a thickness of from about 0.05 mm to about 10,000 micrometers, and typically includes a reflective metallic material such as imprint, silver, gold or steel. Reflective layer ι 4 is typically applied to substrate 112 patterned surface via magnetron spray processing. Another treatment, steaming can be used, but much slower. Chemical wet, plating, enamel treatment (using silver, nickel or other metals) can also be used to form on the patterned surface of the transparent substrate 112. The reflective layer 114. The protective layer 116 typically comprises a lacquer material which, when exposed to ultraviolet (υν) light, is called a "curing" treatment, the material will harden and have a thickness between 1 and 3 microns. The protective layer 116 acts to prevent the reflective layer ι 4 from scratching and oxidizing and to provide a convenient surface on which the label can be printed. The protective layer 丨j 6 is typically applied to the transparent substrate 112 and the reflective layer 114 via a spin coating process, so that when exposed to UV light, a small amount of material is sprayed near the inner diameter of the 988 988 988 988 988 988 988 988 988 988 988 988 988 988 988 988 988 988 988 988 988 988 988 On the disc, and the disc is rotated at a high speed, so that a thin layer of material is overlaid on the surface of the disc. After that, the disc is exposed to UV light, which makes the lacquer material hard. Although a physical layer is included, the optical disc 11 is also considered to have an "information layer, (also, data layer)" 115' at least for the transparent substrate 112 and the reflective layer 114 to contribute to the layer. n information layer, defined by two planes. For this purpose, plane A" is the laser closest to the radial plane and "plane B" is the laser's most distal radial plane, the operational structure extends to this plane. On standard CD or single In a layer of OVD, there is only a single layer of information, and the disc is thus traditionally described as a single data layer (or, a single layer) of the disc. A variety of different CD and DVD standards are expected to have a negligible depth (1.2 mm from the optical axis) and a micro-diameter disc with a radial size of 120 mm. Although the insignificant thickness is 丨·2 mm, the veteran standard for optical disc technology (spoken, "Red Book"), reissued as IEC 908, allows a thickness of 1 3-1.5 mm (for all layer combinations). However, the reader can accommodate one or two additional variations, and a disc suitable for reading by CD and DVD drives can have a maximum of about 2.4 mm with a minimum depth of about 微8 mm, preferably 1·〇-1. 4 mm, more preferably ΐ·υ·3 mm, optimally 1.2 mm. The standard sets a negligible 120 mm in diameter relative to the CD and DVD diameters. However, the optical disc reader can accommodate a disc having a radial diameter of 丨〇〇_丨4〇, preferably 丨1 (M30 mm, more preferably 丨15_125 mm, optimal 988l5.doc 1249158

地為120 mm。甚至於，該標準亦提供具有直徑為8 cm(8〇 mm)的光碟：底座和箝夾環大小保持和i2〇㈤㈤的光碟相 同，/衣度亦相同，只有外徑減低，減低光碟的資料區域。 商業上可使用之CD和DVD閱讀機和讀寫器在個別光碟盤 上容納此直徑的光碟。此種光碟呈現實現本發明的特定優 點，在&些優點間為需要和整個光碟&面接觸而產生效應 的測定樣本容積的等量減低，以及將此種光碟封裝在套筒 大小等於3 1/2”軟性磁碟套筒的該種光碟之能力。 甚至於，各種不同的其他標準如那些定義（磁光）”逑你光 碟”或是類似雷射光碟的標準已經發展，以及其他標準毫 無疑問地將在未來發展。因&，本發明之光碟可以具有徑 向直徑為5。mm之小以及如標準雷射光碟之大，以及可以 調適為在未來發展的標準大小。熟知相關技藝之人士將更 ^ i知A術浯光碟”預期任何合適之可轉動媒體，無 論如何完美地為圓形。 如一般原則，具有4個操作需求必須符合於典型光碟系 、洗正確地仙··該_機必須準確地監督以及控制焦點、 =向位置、切線位置和速度。徑向位置和切線位置的控制 集中一起術語化為尋執。 如上文所討論以及如所示意，光碟之透明基板層 需要將閱讀機雷射適當地聚焦在光碟的反射層ιΐ4上。 失效於維持正確厚度、透 透月性和透明基板層112的折射指 數可能使得反射層114為不可讀取。 步討論’在層115中編碼之操作結構 988l5.doc -21. 1249158 為典型頃取以維持正確尋軌。在標準壓製的CD中，例 如，該閱讀機追蹤壓印在透明基板112上的訊洞螺旋。在 可記錄CD中，該閱讀機追蹤螺旋紋道（在寫入期間，參考 下文）。 " 圖4不意應用在典型cd-類別光碟之氣體_入射、雷射接 近第一表面的非操作結構4〇;該圖使得此種非操作結構將 位於雷射接近入射雷射内定聚焦平面成為明確，該平面為 反射表面114。 # 非操作結構如特定分析物信號元素，如此配置將不可由 標準裝置偵測。 第一，配置在典型CD_類別光碟之氣體-入射、雷射接近 第-表面的非操作結構4〇不僅為雷射接近亦且為正由閱讀 機訊問的外部聚焦平面。這些非操作結構將必須為在可谓 測之入射表面（800 μιη)上光束大小的範圍内。以及最後， 假使為很大時，這些結構4〇由於介入在雷射和反射層Η# 之間的特性，可能干擾讀取在反射表面U4内編碼的操作 結構。因為此後者原因，所以在光碟製造期間經歷實質努 力以確定光碟之雷射接近第一表面實質上沒有瑕疲，部分 為藉由保持在壓模和主片上的表面為乾淨以及沒有灰塵。 因此’為調適標準光碟技術為包含特定分析物信號元素 的非操作結構之目㈣測’具有新光碟和克服這些問題之 光碟製造技術的需要。具有需要於光碟幾何和尋軌設計以 同時和可鑑別地操取由配置在光碟表面上之非操作結構產 生的信號。 98815.doc -22- 1249158 單一資料層光碟 如本發明原理建立之光碟第—系列具體實施例 中術語化為"單一資料層”且興香# y 扣果 而解決這些問題。、”體實細例，藉由展現新的方式 第-，我認知職使該麵作結财質上可以置放 作結構的聚焦平面中時，達成同時聚焦在操作結構和非 作結構的問題將解決，也就是說，實質上和可尋軌社構並、 焦:將欣賞地是該非操作結構和該操作結構不需要在㈣ 的聚焦平面中-足以俊續非j^ & °私作、、，°構和該操作結構為足以 —以允許光碟_機光學讀寫頭❹m二結構。 因為在操作共焦平面上的光束半徑實質上小於在光碟第 一表面的光束半徑·由透明基板⑴折射的結果·次微米之 非操作態樣的使用被許可。 /' 然而’難題是將為可_之非操作結構必須為雷射接近 反射層114 .除非該反射層114為明顯可傳輸（亦即只有部 /刀反射’參考下文），置放於反射層ιΐ4雷射遠端的非操作 結構不比制於標準CD防護層116上的非操作結構為可侦 測。以現存光碟和製造處理而言，該非操作結構將必須置 放於透明基板112和反射層114之間以將它們置放為雷射接 近於反射層114以及實質上和反射層m共焦。此種解答， 強制反射層m在非操作結構配置之後的單獨化應用，於 實現光碟為部分快速特定分析物驗定時不可保持。 在此問題的新解答中’由於標準單一資料層cd_類別光 碟之翻轉實體方位的原因（換句話說，在閱讀機中上下顛 98815.doc -23- 1249158 倒置放），將呈現其他雷射遠端表面為光碟之雷射接近表 面。 如圖5A和5B所示意，將傳統CD_類別光碟翻轉，是將防 護層116呈現為光碟的雷射接近第一表面。假使防護層ιΐ6 被移除，非操作結構136之後實質上可以置放為與反射層 114共焦以及雷射接近。 光碟的另一實體翻轉-幾何上，相對於徑向平面（如那些 由此定義的術語）繪畫的切線大約丨8〇。之轉動-使二重大改 變產生效應。 第一種是實體倒置：在標準光碟中合適地置放以用於光 碟閱讀機讀取之雷射接近的結構（亦即平面”A”）變成雷射 遠端（亦即移動為平面” B")，而且標準光碟中合適地置放以 用於光碟閱讀機讀取之雷射遠端的結構（亦即平面"B")變成 雷射接近（亦即移動為平面"A")(在圖5A_5B中示意）。第一 種是"邏輯"或是"影像”倒置：由光碟機光學拾音器（目標组 件)的觀點’由光碟操作結構建立的幾何形式顯現為映射 影像-也就是說，像是反射跨越在徑向平面中繪晝的切線 (如由此定義）反射（在圖5C-5D中示意）。 該實體倒置呈現較少困難度··如下文更進一步討論，我 已經發現如CD-R標準之沒有染料的擺動"紋道"可以非常容 易地尋軌，無論該擺動"紋道"為雷射接近或是雷射遠端浮 雕0 幾何形式在其映 該額外，，影像倒置 然而，當操作結構（特別是尋執）建立的 射影像（亦即，對掌性）上為不可重叠時， 98815.doc -24- 1249158 ==傳統光碟閱讀機中為不可讀取”。此後者之問題 / 由傳統用於以⑶格式尋軌的《加以舉例證 明0 機二VI:先碟閱讀機逆時鐘方向旋轉光碟，如由光碟 !t/" 來看’以及由光碟内徑⑽螺旋至光碟 二(：d.)。該螺旋因此由光學拾音器觀點顯現，如圖 :。*該光碟翻轉以及由反側看時，有效地反射螺 疋 向平面的切線上，該螺旋如圖5C所示意加以顯 現，以及不能在傳統光碟機中由u.至〇.d尋軌。 ^更進-步討論的目的，該片語"前向影像"由此用以來 ^由光碟操作結構建立之幾何形式由光碟機光學拾音器 來看在#向平面十的傳統方位。特別是，該片語由此用以 參考為由光碟可尋軌結構建立之幾何形式由光碟機光學拾 音_在控向平面中的傳統方位。當應用於光碟時，該 片語，此用以參考為由光碟可尋軌結構本身建立之幾何形 式或疋由此參考為投射影像。 該f吾”反向影像"參考為反方位-也就是說，參考為藉由 反射則向影像在徑向平而φ彡合查 中繪畫的切線上而獲得的方位。 類似地是，該片語"正浮雕"將由此參考為操作結構在光 軸（介於資訊層平面，，A"和"B"之間）方向之傳統實體方位， 由光碟機光學拾音器的關點來看。特別是，該片語"正浮 雕”參考為光碟讀取期間尋執之操作結構的傳統實體方 位，由光碟機光學拾音器的關點來看。當應用於光碟時， 該片语參考為結構本身。 98815.doc -25- !249158 f片5吾，負浮雕"將參考為相反實體方位。 ·】當由此使用時’該片語"標準主片"參考為影像和浮雕典 型地用Μ在傳統處理中產生標準光碟之主片光碟。該片語 反向主片"視為具有影像，但不具浮雕，與傳統相反之主 本發明為部分依據非操作結構可以容易地與光The ground is 120 mm. Even the standard provides a disc with a diameter of 8 cm (8 〇mm): the size of the base and the clamp ring is the same as that of the i2 〇 (5) (5), and the clothing is the same, only the outer diameter is reduced, and the data of the disc is reduced. region. Commercially available CD and DVD readers and readers accommodate discs of this diameter on individual discs. Such a disc exhibits a particular advantage in achieving the present invention, between the & advantages, an equivalent reduction in the volume of the assay sample that is required to produce an effect with the entire disc & face contact, and packaging the disc at a sleeve size equal to 3 The ability of the 1/2" flexible disk sleeve for this type of disc. Even various other standards such as those defined (magneto-optical) "逑 your disc" or similar laser-disc have been developed, as well as other standards. Undoubtedly, it will develop in the future. Because of &, the optical disc of the present invention can have a radial diameter of 5 mm and as large as a standard laser disc, and can be adapted to a standard size for future development. Those skilled in the art will be more aware of any suitable rotating media, in any case perfectly circular. As a general rule, having four operational requirements must be consistent with a typical disc system, washing the correct ones. The machine must accurately monitor and control the focus, = position, tangent position and speed. The control of the radial position and the tangential position is collectively termed as a search. As discussed above and as illustrated, the transparent substrate layer of the optical disc requires proper focus of the reader laser on the reflective layer ι4 of the optical disc. Failure to maintain the correct thickness, permeability, and refractive index of the transparent substrate layer 112 may render the reflective layer 114 unreadable. Step Discussion 'Operation Structure Encoded in Layer 115 988l5.doc -21. 1249158 is a typical one to maintain proper tracking. In a standard pressed CD, for example, the reader tracks the hole spiral imprinted on the transparent substrate 112. In a recordable CD, the reader tracks the spiral track (during writing, see below). " Figure 4 is not intended to be applied to the gas-injection of a typical cd-class disc, the non-operational structure of the laser approaching the first surface; this figure makes this non-operational structure will be located near the incident focal plane of the laser. It is clear that the plane is the reflective surface 114. # Non-operational structures such as specific analyte signal elements, such configuration will not be detectable by standard equipment. First, the non-operating structure 4 of the gas-incident, laser-proximate first-surface configuration of a typical CD-class disc is not only a laser approach but also an external focus plane that is being interrogated by the reader. These non-operational structures will have to be within the range of beam sizes on the measurable incident surface (800 μηη). And finally, if it is large, these structures may interfere with the reading of the operational structure encoded in the reflective surface U4 due to the nature of the interaction between the laser and the reflective layer Η#. For this latter reason, substantial efforts are made during the manufacture of the optical disc to determine that the laser of the optical disc is substantially free of fatigue near the first surface, in part by keeping the surface on the stamp and the main sheet clean and free of dust. Therefore, the need to adapt standard optical disc technology to a non-operational structure containing specific analyte signal elements (4) has the need for new optical discs and optical disc manufacturing techniques that overcome these problems. There is a need for a disc geometry and tracking design to simultaneously and identifiably manipulate signals generated by non-operating structures disposed on the surface of the disc. 98815.doc -22- 1249158 A single data layer optical disc is created according to the principle of the present invention. In the specific embodiment, the term "single data layer" and "Xingxiang #yy" are used to solve these problems. By way of example, by presenting a new way - I recognize that the face can be placed in the focal plane of the structure, and the problem of achieving both the operational structure and the non-structural structure will be solved. That is to say, in essence, and the track-searchable organization, the focus is that the non-operational structure and the operational structure do not need to be in the focal plane of (4) - enough to continue the non-j^ & ° private work, ° The structure and the operational structure are sufficient to allow the optical disc to be optically read and written. Since the beam radius on the operating confocal plane is substantially smaller than the beam radius at the first surface of the disc, the result of the refraction of the transparent substrate (1), the use of the non-operating aspect of the sub-micron is permitted. /' However, the problem is that the non-operating structure must be a laser close to the reflective layer 114. Unless the reflective layer 114 is clearly transmissive (ie, only the part/knife reflection 'see below), placed on the reflective layer The non-operating structure of the ιΐ4 laser distal end is not detectable than the non-operating structure fabricated on the standard CD protection layer 116. In the case of existing optical discs and fabrication processes, the non-operational structure will have to be placed between the transparent substrate 112 and the reflective layer 114 to place them in proximity to the reflective layer 114 and substantially confocal to the reflective layer m. In this solution, the separate application of the forced reflection layer m after the non-operational structure configuration is not maintained for the implementation of the optical disc for a part of the fast specific analysis. In the new answer to this question, 'because of the flipping physical orientation of the standard single data layer cd_ category disc (in other words, upside down in the reader 98815.doc -23-1249158), other lasers will be presented. The distal surface is the laser of the disc approaching the surface. As illustrated in Figures 5A and 5B, the conventional CD_class disc is flipped over by exposing the protective layer 116 as a disc with the laser approaching the first surface. If the protective layer ι 6 is removed, the non-operating structure 136 can then be placed substantially confocal with the reflective layer 114 as well as the laser. Another entity of the disc flips - geometrically, a tangent to the plane of the drawing relative to a radial plane (such as those defined by this) is approximately 〇8〇. The rotation - causing two major changes to produce an effect. The first is physical inversion: the structure that is properly placed in a standard disc for the laser reader to read close to the laser (ie, plane "A") becomes the far end of the laser (ie, moves to a flat surface) B&quot ;), and the structure of the remote end of the laser that is properly placed in the standard disc for reading by the optical disc reader (ie, plane "B") becomes laser close (ie, moving to plane "A") (Illustrated in Figures 5A-5B.) The first is "logic" or "image" inversion: from the perspective of the optical pickup (target component) of the optical disk, the geometric form established by the optical disk operating structure appears as a mapped image - That is, reflections such as reflections across a tangent (as defined thereby) drawn in a radial plane (illustrated in Figures 5C-5D). The inversion of the entity presents less difficulty. As discussed further below, I have found that the sway of the dye-free "striker" as in the CD-R standard can be very easily tracked, regardless of the swing "grain" For the laser approaching or the laser remote embossing 0 geometry form in which it is reflected, the image is inverted, however, when the operational structure (especially the seek) establishes the image (ie, the palm of the hand) is not When overlapping, 98815.doc -24- 1249158 == is not readable in traditional CD-ROM readers. This latter problem / by traditionally used to track in (3) format, to exemplify 0 machine two VI: first disk reading Rotate the disc counterclockwise, as seen by the disc!t/" and spiral from the inner diameter of the disc (10) to the disc 2 (:d.). The spiral is thus visualized by the optical pickup, as shown in the figure: * When flipping and looking from the opposite side, it effectively reflects the tangential line of the snail to the plane. The spiral is visualized as shown in Fig. 5C, and cannot be tracked by u. to 〇.d in a conventional optical disc drive. The purpose of the discussion, the phrase "forward image "The resulting geometric form of the optical disc operating structure is viewed by the optical pickup of the optical disc machine in the conventional orientation of the plane to the plane. In particular, the phrase is used for reference to establish a track-retrievable structure from the disc. The geometric form is optically picked up by the optical disc drive_the conventional orientation in the control plane. When applied to a disc, the phrase is used to refer to the geometric form established by the disc trackable structure itself or by reference. To project an image, the reference is a reverse orientation—that is, the reference is the orientation obtained by reflection to the tangent of the image in a radial plane and a φ 彡 查 。 Similarly, the phrase "positive relief" will be referred to as the traditional physical orientation of the operational structure in the direction of the optical axis (between the information layer plane, A" and "B"), by optical disc optics Look at the pickup point. In particular, the phrase "positive relief" refers to the traditional physical orientation of the operational structure of the optical disk during reading, as seen by the point of view of the optical pickup of the optical disc. When applied to a disc, the phrase reference is a structure. 98815.doc -25- !249158 f-five 5, negative relief " will refer to the opposite physical orientation. ·] When used as such, the phrase "standard master" " reference for images and reliefs typical The main film used to produce a standard optical disc in a conventional process. The phrase reverse main film " is considered to have an image, but is not embossed, contrary to the tradition, the present invention is partially based on a non-operational structure that can be easily Light

f共焦配置的新發現，以及另外雷射接近於先前應用U ^ °此為藉由製造光碟完整基板（也就是說，該基板表 ^足成光碟的資訊層）圖樣化表面呈現為基板最雷射接近 表面（此後稱為，，倒置光碟”）而完成。 1我已經發現此種光碟可以藉由使用傳統母片為壓模而製 ^因此產生_作結構以正浮雕呈現的前向影像倒置光碟 (”處理1”）。第二製造處 直 八笮乂片或疋子片使用為壓模 亦加以說明。扁品牮_ # Λ丄 在此第一處理中，該主片具有與傳統反向的 影像；該形成的前向影像倒置光碟具有以負浮雕呈現之操 作結構。 雖然上述之新方式界決同時建立焦點在非操作結構和操 作結構上的光學問題，但它們本身不需要允許此種結構的 同時谓測。第…這些方式不需要解決維持此種共焦配置 呈現時非操作結構的尋軌、聚焦和同步（速度控制）問題。 第二，與操作態樣（例如訊洞）共焦配置的非操作結構可能 造成信號不足以由操作(以及資料編碼)信號區別以由此防 止鑑別。 生的電子信號較先前 現在我已經發現在光碟讀取期間產 98815.doc -26- 1249158 已經瞭解地具有更大的資訊内容。也就是說，該電子信號 為設計以提供二特定類別資訊-用於光碟操作的必要資訊 以及數位式編碼在光碟上的資訊_事實上考以用各種方法 查詢以及量測以揭露先前有關光碟實體結構的未開發之大 量豐富資訊。 甚至於，我已經發明允許這些信號之所需操作分量被分 離的方式，除去該#唬其餘部分而不干擾成功的驅動操 作。在本發明此特徵之一具體實施例中，新光碟製造為其 中光碟操作所需的信號，特別是光碟尋執所需的信號在讀 取期間由用以報告在光碟上非操作結構反應的信號部分分 離，而允許在倒置光碟非操作結構的同時和可鑑定偵測。 因此，在本發明此特徵之第一系列具體實施例中，光碟 呈現為允許尋執信號由HF(RF，四列總和）信號分離，除去 HF彳。號以報告在非操作結構上配置的資訊。尋執信號由 HFk號的目的分離為使用尋執設計完成，由現存光碟標準 在光碟讀取期間意圖忽略，該擺動，，紋道，，制訂在^〇和 可寫入以及可再寫入標準中。 5亥擺動’’紋道’’首先加入至光碟標準以允許使用者-導引記 錄（寫入）至CD(以及後來的DVD)媒體中。如在相關標準中 所陳述，口語術語化為”橘皮書，，，該擺動在使用者寫入資 料至光碟期間單獨由記錄裝置偵測；此後，尋執為藉由偵 測身料沿著螺旋寫入而完成。該標準以及標準的所有現存 製作因此預期和教導該擺動變成冗餘以及因此在寫入之後 忽略。 988l5.doc 27- 1249158 雖然由現存標準在渠取期間意圖忽略，該擺動紋道呈現 一些重大優點為用於在本發明倒置光碟上的尋軌設計。 和螺旋訊洞相反，該擺動提供HF中相當低頻信號。因 為非操作結構（如特定分析物信號元素），在此由該結構舉 例4明為0.5-10微米大小範圍，該擺動使用為尋軌結構允 許由尋軌信號的頻率為基礎之非操作分離。 非操作結構發生的較高頻HF事件之一結果為可以容易 地摘測和特徵化。藉由舉例方式，圖13為數位示波器顯示 在特疋免疫附著在本發明前向影像、正浮雕倒置光碟之雷 射接近第一表面上紅血球細胞上可尋執掃描期間所獲得 卵（四列總和，RF)信號中之擾亂，其中χ軸顯示時間而γ 軸顯示四列總和信號大小。 此種頻率為基礎之分離的第二、相反結果為該非操作信 發生之更高頻信號為夠短歷時以防止尋執鎖定之遺漏。 甚至於，低通或是凹槽濾波器可用以由偵測器“報告之信 號移除該高頻事件，更進一步確定正確尋執。將欣賞地是 該濾波器由用以_非操作信號（如特定分析物信號）的伴 隨信號部分省略。 擺動紋道於本發明光碟用為尋軌設計的另一主要優點為 是擺動信號可由該驅動裝置使用以在光碟上所有點維持常 數線性掃描速度。既定已知擺動頻率，有關高頻非操作事 件的大小資訊可以因此容易地計算，如例子3所顯示以及 在下文更進一步討論。相反地，該光碟可用具有至少—選 擇擺動頻率之紋道加以mastered ’而允許光碟線性速度為 98815.doc -28- 1249158 徑向平面之任何既定點加以指定。The new discovery of f-focal configuration, and the additional laser is close to the previous application U ^ °. This is by making the complete substrate of the optical disc (that is, the information layer of the substrate is formed into a disc). The laser is completed close to the surface (hereinafter referred to as, inverted optical disc). 1 I have found that such a disc can be made by using a conventional master for stamping, thus producing a forward image of the structure with a positive relief. Invert the disc ("Process 1"). The second manufacturing section of the straight gossip or scorpion sheet is also used as a stamper. Flat 牮 _ # Λ丄 In this first process, the main piece has a tradition Inverted image; the formed forward image inverted optical disc has an operational structure presented in a negative relief. Although the new method described above simultaneously establishes optical problems of focus on the non-operational structure and the operational structure, they do not need to allow themselves. Simultaneous prediction of such a structure. The first method does not need to solve the problem of tracking, focusing and synchronization (speed control) of the non-operational structure when maintaining such a confocal configuration presentation. A non-operational structure of a confocal configuration (such as a hole) may cause a signal to be insufficient to be distinguished by an operational (and data encoding) signal to thereby prevent discrimination. The raw electronic signal is earlier than I have now found during the optical disk reading. .doc -26- 1249158 already knows that there is more information content. That is, the electronic signal is designed to provide two specific categories of information - the necessary information for the operation of the disc and the digitally encoded information on the disc - facts The test was conducted in various ways to inquire and measure to reveal a large amount of untapped information about the physical structure of the optical disc. Even I have invented the way to allow the required operational components of these signals to be separated, removing the rest of the #唬Without disturbing the successful driving operation. In one embodiment of this feature of the invention, the new optical disc is manufactured as a signal required for the operation of the optical disc, and in particular, the signal required for the optical disc seek is reported during reading. The signal on the disc is separated from the non-operating structure, and the signal is allowed to be inverted while inverting the non-operating structure of the disc. Therefore, in a first series of embodiments of this feature of the invention, the optical disc is presented to allow the seek signal to be separated by the HF (RF, Four Columns Sum) signal, removing the HF 彳. to report on the non-operational structure. The configuration information. The search signal is separated by the purpose of the HFk number to be completed using the search design. The existing optical disc standard is intended to be ignored during the reading of the optical disc. The swing, the groove, is formulated in the ^ and writable and can be Re-written in the standard. 5 Hai swing ''slot'' is first added to the disc standard to allow user-guided recording (writing) to the CD (and later DVD) media. As stated in the relevant standards The colloquial term is "Orange Book," and the swing is detected by the recording device alone during the writing of the data to the disc by the user; thereafter, the seek is completed by detecting the body along the spiral writing. This standard, as well as all existing productions of the standard, therefore anticipates and teaches that the wobble becomes redundant and therefore ignored after writing. 988l5.doc 27-1249158 Although it is intended to be ignored during the dredging by existing standards, the wobble track presents some significant advantages for the tracking design on the inverted optical disc of the present invention. In contrast to a spiral hole, this swing provides a relatively low frequency signal in the HF. Because of the non-operational structure (e.g., a particular analyte signal element), the structure of Example 4 is here considered to be in the range of 0.5-10 microns, and the wobble is used as a tracking structure to allow non-operational separation based on the frequency of the tracking signal. One of the higher frequency HF events that occur with non-operational structures results in easy sniffing and characterization. By way of example, Figure 13 is a digital oscilloscope showing the eggs obtained during the scannable scan of the red blood cells on the first surface of the laser that is attached to the forward image and the embossed inverted optical disc of the present invention. , RF) The disturbance in the signal, where the x-axis shows time and the γ-axis shows the four-column sum signal size. The second, opposite result of such frequency-based separation is that the higher frequency signal generated by the non-operational signal is short enough to prevent omission of the seek lock. Even a low pass or groove filter can be used to remove the high frequency event by the detector's "reported signal, further determining the correct seek. Appreciate that the filter is used by the _ non-operational signal ( The accompanying signal portion of the specific analyte signal is omitted. Another major advantage of the wobble track for the track design of the present invention is that the wobble signal can be used by the drive to maintain a constant linear scan speed at all points on the disc. Given the known wobble frequency, the size information about the high frequency non-operational events can thus be easily calculated, as shown in Example 3 and discussed further below. Conversely, the disc can be mastered with a track having at least a select wobble frequency. 'Allow the linear speed of the disc to be specified by any predetermined point in the radial plane of 98815.doc -28- 1249158.

除此之外，已知為雙相標示信號的另一信號為編碼在 CD-R光碟上的擺動信號頂端。該雙相標示信號提供可用 以決定光碟上位置的位址資訊。此位址資訊有利於特定分 析物驗定之立體-可尋址特徵；該空間可定址性之效用為 更進步在共同擁有及尚在審理申請案之1997年7月7曰申 請的第〇謂8,935號；1998年7月21日中請，第〇9/12〇,〇49 號中加以說明，該文獻以提及的方式併入本文中。在 DVD-R格式巾’類似資訊為使用"紋面預製訊洞"加以提 供，因此在紋面區域（介於擺動紋道之間）中的預製沖壓凹 槽將位址資訊編豸。在用於本發明之CD或是dvd格式 令，該雙相標示或是紋面預製訊洞編碼可以選擇性地 定區域省去。 雖然特別參考非操作信號由操作信號分離，本發明預期In addition to this, another signal known as a bi-phase indicator signal is the top of the wobble signal encoded on the CD-R disc. The bi-phase indicator signal provides address information that can be used to determine the location on the disc. This address information facilitates the stereo-addressable characteristics of a particular analyte; the space-addressable utility is more advanced in the co-owner's application of the application of July 7th, 1997. No. 9/12 〇, 〇 49, which is incorporated herein by reference. In the DVD-R format, the similar information is provided using the "textured prefabricated hole", so the pre-formed stamping grooves in the textured area (between the wobbled tracks) encode the address information. In the CD or dvd format used in the present invention, the two-phase label or the grain prefabricated hole coding can be selectively omitted. Although the reference to the non-operational signal is specifically separated by the operational signal, the present invention contemplates

能夠具有依據CD及/或DVD標準數位式編碼之資訊的光 碟；=此陳述之方式允許該數位式編碼之資訊以非操作結 構的讀取加以回復。 以及雖然特別參考HF信號内的頻率分離加以說明 發明預㈣測器產生之不同於四列總和(如尋軌誤 ::其他類別信號之分離和組合可用以偵測及特徵化非操作 曰刖只頁光碟撰 、 吻^能勒之能力。 因為該擺動典型只在寫入期間被偵測， 制A ^ 尸坏以本發明大多翁 製乍不#要以及造成雷射可能以干擾非操作結構们則之振 98815.doc -29- 1249158 幅發生脈衝之處理，特殊設計用於品質控制目的之讀寫器 在例子2及例子3中利用。此裝置能夠偵測及尋執擺動而不 會強制雷射以光碟寫入所需的能量發生脈衝。然而，將瞭 解地是，能夠偵測該尋執結構之驅動器，無論是否能夠寫 入，都可以利用於本發明之實現中。 由此呈現之例子2及例子3在下文實際說明本發明單一資 料層、前向影像正浮雕倒置光碟之成功利用以（1)藉由免疫 分析偵測人體血液中之IgG，以及（2)使用最小修正光碟閱 讀機彳貞測和特徵化在光碟表面上捕捉之人體紅血球。例子 7實際說明核酸基礎驗定之調適於由此說明的偵測原理。 單一資料層光碟製造 用於製造本發明單一資料層、前向影像光碟之第一處理 (處理Γ’）現在將參考圖3 A-3D加以說明。處理1形成具有 正浮雕之操作結構的光碟。 透明光碟基板112可以藉由注入鑄造處理加以製造，其 中熔解之聚碳酸脂被注入至具有”壓模”之鑄造孔穴中。該 壓模藉由電鑄以光碟主片製成，將在下文更完全地說明。 該注入鑄造處理典型地需每光碟花費5至丨〇秒。 圖3 A示意在標準電鑄和基體化處理中之3步驟，緊接著 為最終注入鑄造（沖壓）步驟，處理（沖壓）步驟，用於建立 具有擺動紋道之單一資料層光碟的完整基板112，CD-R或 是CD-RW光碟（更進一步細節參考WilliamsA撰寫，"The CDjlROM and Optical Disc recording SvstemsM，牛津出版 社’ 1994年，在此處以提及的方式併入本文中）。主片 98815.doc -30· 1249158 51〇、父片520、母片530、子片540和光碟基板112以側面 橫斷面部分圖式加以顯示。 簡短地是，該處理可以主片光碟51〇之建立開始（由玻 璃、塑膠等製成），該步驟在圖3八顯示之那些步驟之前進 仃。在步驟505中，主片光碟51〇被置放於電流鎳電解質槽 中，该光碟在該處作用為電子電路之陰極。鎳陽極為用以 將層鎳沈澱在主光碟表面上，建立錄，，父片"52〇。接 著，鎳父片520由主片510分離，典型地（但不是總是）在處 理中破壞主片510。結果，父片520以主片51〇之負浮雕相 反影像浮凸壓印。 雖然父片52〇可以在注射鑄造處理535中直接使用為壓 模但已經證實在相關技藝中為更有效以產生父片520之 數相專複製，術5吾化為”子片” $ 4 〇。由原始錄部分建立 之電鑄部分家族術語化為"基體化，，。 因此，父片520典型地在步驟5丨5中電流地使用以串列產 生多數相等之”母片”530。母片53〇具有以正浮雕呈現之前 向衫像。該多數相等母片53〇，依次，在步驟525中用以產 生多數子片540，影像方位和浮雕等於，，父片”52〇，以及因 此具有和主片光碟5丨〇比較為負浮雕呈現之反向影像。在 v驟535中，子片540使用為光碟基板1丨2之注射鑄造的壓 模如此製造之光碟具有以正浮雕呈現之前向影像。 圖3C顯不具有如圖3A製造之基板112的CD_R光碟u，具 有擺動紋道118、反射層114和防護層ιι6。在擺動”紋道” 上的光學路徑和焦點以虛線顯示。如所能瞭解，在典型光 98815.doc -31 - 1249158 碟之基板112以其雷射接進至其圖樣化表面之平面表面呈 現0 圖3B顯示如本發明之一處理（此後稱為”處理厂），用於 建立本發明具有以正浮雕呈現的操作結構之前向影像的單 為料層倒置光碟。該處理和圖3入顯示之標準處理不同， 在該處理中，母片53〇在步驟625中直接使用以沖壓光碟基 板132。如例子1和例子5所陳述，在此報告之實驗中所使 用的光碟為使用此種修正之基體化程序以在ΕχΐΜρ〇 S.R.〇(布拉格，捷克共和國）產生。 處理1之一優點為標準主片-如。仏尺主片或是dRw主 片-或是另外容易使用現存技術和裝置建立之主片，可用 以製造本發明前向影像倒置光碟。 在處理1中，如圖3B和例子丨和例子5中所示意，母片以 標準注射鑄it機詩放直接纽射鑄料理之錢發生某 些程度之啟始困難度。注射鑄造機器之各種不同參數對那 些熟知相關技藝之技士將是明顯的，將被調整以致於溶化 的聚碳酸脂在鑄模上適當地流動。例如，聚碳酸脂注入至 之溫度被提升以確；t熔化的聚碳酸脂為較不黏。雖然此增 加之溫度可能改變聚碳酸脂光學性質(例如雙折射），聚碳 酸脂層132之光學性質對本發明單一資料層倒置光碟效能 為無形的’因為雷射從不通過此資料層。圖4ΐΑ·川呈現 使用於如例子5中所陳述製造之光碟製造的鑄模設定。 圖3D顯示由處理！形成之光碟，而非操作結構136與反射 層U4共焦應用以及雷射接近於反射層134。如所能瞭解， 988 丨 5.doc -32- 1249158 在本發明單一貧料層倒置光碟中之基板丨32以其圖樣化表 面雷射接近於其平面表面而加以呈現。如亦所能瞭解，如 此製造之光碟具有以正浮雕呈現之前向影像。 將理所當然瞭解地是倒置光碟所謂，，平面表面，，不需要非 常平面，因為不位於光學路徑中。因此，此表面由此亦將 術語化為光碟基板之，，額外”表面。 在一標準光碟和本發明前向影像/正浮雕倒置光碟尹， 正尋執之結構-該擺動"紋道為在資訊層平面"A"中，也就 是說，分別在圖3C和圖3D所示之資訊層雷射接***面 中。 然而，在標準光碟中，平面"A”為最接近於該基板平面 表面之資訊層平面。相反地，在本發明前向影像/正浮雕 倒置光碟中，平面”A”為該基板平面表面之資訊層最遠端 平面。此後者結構方位之結果，以及儘管說明尋軌結構之 術語”紋道"的π語化使用，我已經發現配置在單—資料層 前向影像/正浮雕光碟之反射層上的非操作結構136傾向於 將它們本身定在鄰近尋軌結構本身中央而非落在尋軌結構 本身内，如圖3D所示。 用於製k本發明單一資料層、前向影像倒置光碟之第二 处里（此後稱為處理2 ’’）現在將加以說明。使用處理2製造 之光碟具有操作結構以負浮雕呈現之前向影像。 在處理2中’主片光碟建立為具有與傳統相反之所需操 作〜構的衫像。基本上如圖3 A所示，緊接著傳統電鱗和基 體化處理之後，當以其圖樣化表面雷射接進制其平面表面 98815.doc -33- 1249158 (亦即倒置）而讀取時，該光碟基板具有操作態樣為負浮雕 之前向影像。 因此，和標準光碟（圖3C中所示）以及如處理丨製造之本 發明光碟（圖3D中所示）相反，如處理2製造之本發明光碟 中所示）具有在平面” B"之尋軌結構，也就是說，在 、貝訊層雷射遠端平面中。在這些倒置光碟卡之平面為A disc capable of having digitally encoded information in accordance with CD and/or DVD standards; = this statement allows the digitally encoded information to be replied with a non-operational read. And although the frequency separation in the HF signal is specifically referred to, the invention produces a pre-(four) detector that is different from the four-column sum (such as tracking error:: separation and combination of other types of signals can be used to detect and characterize non-operational 曰刖The ability to slap, kiss, and sing. Because the sway is typically detected only during the writing process, the A ^ corpse is broken by the majority of the invention and the laser may interfere with the non-operating structures. Then the vibration of 98815.doc -29- 1249158 is processed by pulse. The reader specially designed for quality control is used in examples 2 and 3. This device can detect and seek swing without forcing thunder. The pulse of energy required for the writing of the optical disc is generated. However, it will be appreciated that the driver capable of detecting the homing structure, whether or not it can be written, can be utilized in the implementation of the present invention. 2 and Example 3 The following is a practical description of the successful use of the single data layer and forward image positive relief inverted optical disc of the present invention to (1) detect IgG in human blood by immunoassay, and (2) use minimal correction The optical disc reader speculates and characterizes the human red blood cells captured on the surface of the optical disc. Example 7 actually illustrates that the adjustment of the nucleic acid base assay is suitable for the detection principle thus described. Single data layer optical disc manufacturing is used to fabricate a single data layer of the present invention, The first process (process Γ') of the forward video disc will now be described with reference to Figures 3A-3D. Process 1 forms a disc having a positive relief operating structure. The transparent disc substrate 112 can be fabricated by injection casting processing, wherein The molten polycarbonate is injected into a casting cavity having a "die" which is made by electroforming in a master disc of the optical disc, which will be more fully explained below. The injection casting process typically requires a cost per disc. 5 to leap seconds. Figure 3A shows the 3 steps in the standard electroforming and matrixing process, followed by the final injection casting (stamping) step, the processing (punching) step, for creating a single data with a wobbled track. The complete substrate 112 of the layered disc, CD-R or CD-RW disc (for further details, refer to Williams A, "The CDjlROM and Optical Disc recording Svstems M, Oxford Press '1994, incorporated herein by reference.) Main film 98815.doc -30· 1249158 51〇, parent piece 520, mother piece 530, sub-piece 540, and optical disk substrate 112 The cross-section of the side cross-section is shown. Briefly, the process can begin with the creation of the master disc (made of glass, plastic, etc.), which step is preceded by those shown in Figure 38. In step 505, the main piece of the optical disc 51 is placed in a current nickel electrolyte bath, where the optical disc acts as a cathode of the electronic circuit. The nickel anode is used to deposit the layer of nickel on the surface of the main optical disc to establish a recording. Parent film " 52 〇. Next, the nickel parent sheet 520 is separated by the main sheet 510, typically (but not always) destroying the main sheet 510 in the process. As a result, the parent piece 520 is embossed with the negative relief of the main piece 51〇. Although the parent piece 52 can be used directly as a stamp in the injection casting process 535 but has been shown to be more effective in the related art to produce a digital copy of the parent piece 520, the process is "sub-slice" $4 〇 . The family name of the electroformed part established by the original recorded part is changed to "base,. Thus, parent tile 520 is typically used galvanically in steps 5丨5 to produce a plurality of equal "masters" 530 in series. The mother piece 53 has a front jersey image in a positive relief. The majority of the equal masters 53〇, in turn, are used to generate a plurality of sub-slices 540 in step 525, the image orientation and the relief are equal to, the parental film "52", and thus has a negative relief representation compared to the main disc 5's. The reverse image. In v 535, the sub-slice 540 is fabricated using a shot-casting die for the optical disk substrate 1 丨 2, and the optical disk thus produced has a forward embossed front image. Figure 3C shows that it is manufactured as shown in Fig. 3A. The CD_R disc u of the substrate 112 has a wobble track 118, a reflective layer 114 and a protective layer ι6. The optical path and focus on the wobble "track" are shown in dashed lines. As can be seen, in the typical light 98815.doc -31 - 1249158 The substrate 112 of the dish is rendered with its laser attached to the planar surface of its patterned surface. FIG. 3B shows one of the processes of the present invention (hereinafter referred to as "processing plant") for establishing the present invention with positive relief The operating structure is previously used to invert the disc to a single layer of the image. This process is different from the standard process shown in Figure 3, in which the master 53 is used directly in step 625 to stamp the disc substrate 132. As stated in Examples 1 and 5, the optical discs used in the experiments reported herein were produced using the modified matrixing procedure in ΕχΐΜρ〇 S.R.〇 (Prague, Czech Republic). One of the advantages of Process 1 is the standard master film - such as. The main film or the dRw main film - or another main piece that can be easily built using existing technologies and devices, can be used to manufacture the forward video inverted optical disc of the present invention. In the process 1, as shown in Fig. 3B and the example 丨 and the example 5, the master piece has a certain degree of difficulty in starting the money of the standard injection molding machine. The various parameters of the injection casting machine will be apparent to those skilled in the art and will be adjusted so that the melted polycarbonate flows properly over the mold. For example, the temperature at which the polycarbonate is injected is increased to ensure that the molten polycarbonate is less viscous. While this increased temperature may alter the optical properties of the polycarbonate (e.g., birefringence), the optical properties of the polycarbonate layer 132 are intangible to the single data layer inversion of the present invention' because the laser never passes through the data layer. Fig. 4 shows the mold setting used for the manufacture of the optical disc manufactured as set forth in Example 5. Figure 3D shows by processing! The formed optical disc, rather than the operational structure 136, is in confocal application with the reflective layer U4 and the laser is adjacent to the reflective layer 134. As can be appreciated, 988 丨 5.doc - 32 - 1249158 The substrate 丨 32 in the single poor layer inverted optical disc of the present invention is presented with its patterned surface laser approaching its planar surface. As can also be appreciated, the optical disc thus manufactured has a forward embossed forward image. It will of course be understood that the inverted optical disc is called a flat surface, and does not require a very flat surface because it is not located in the optical path. Therefore, this surface is thus also termed into a disc substrate, an additional "surface. In a standard optical disc and the present invention, the forward image/positive embossed inverted disc Yin, the structure being sought - the swing " In the information layer plane "A", that is, the information layer lasers shown in Figures 3C and 3D, respectively, are close to the plane. However, in the standard optical disc, the plane "A" is the closest to the substrate. The information layer plane of the planar surface. In contrast, in the forward video/positive embossed optical disc of the present invention, the plane "A" is the farthest plane of the information layer of the planar surface of the substrate. The result of the structural orientation of the latter, and although the π-lingualization of the term "grain" is used to describe the tracking structure, I have found that the non-operational structure is placed on the reflective layer of the single-data layer forward image/positive embossed disc. 136 tends to position themselves in the center of the tracking structure itself rather than falling within the tracking structure itself, as shown in Figure 3D. For use in making the second data layer of the present invention, the second portion of the forward image inversion disc (hereinafter referred to as Process 2 '') will now be explained. The disc manufactured using Process 2 has an operational structure to present the forward image to the negative relief. In Process 2, the 'Master CD is established to have the desired operation opposite to the conventional one~ The structure of the shirt. Basically as shown in Figure 3A, immediately after the traditional scale and substrate treatment, when the surface of the surface is laser-like, the plane surface is 98815.doc -33-1249158 (that is, inverted) And when read, the optical disc substrate has an operational aspect of a negative embossed forward image. Therefore, contrary to a standard optical disc (shown in FIG. 3C) and the optical disc of the present invention (shown in FIG. 3D) manufactured by processing 丨As shown in the optical disc of the invention manufactured by Process 2, it has a tracking structure in the plane "B", that is, in the far-end plane of the laser beam. On the plane of these inverted disc cards is

光碟基板平面表面最遠端資訊層之平面^結果，配置在反 射表面上的非操作結構將經f定在尋軌態樣上中心。此情 形應該期望地增加信號在由定於該鄰近磁軌中^之相同非 操作結構可獲得信號之上。 ^本^明則向影像/正浮雕倒置光碟（處理丨）或是前向影 像:負浮雕倒置光碟(處理2)而言，反射層134現在呈現為光 第I面應.亥疋明顯的’沒有透明基板層η 2呈現以協 助雷射聚焦’如標準光碟。圖6Α以前向影像/正浮雕倒置 光碟解釋問題。The plane of the farthest information layer on the plane surface of the disc substrate, the non-operating structure disposed on the reflecting surface will be centered on the tracking pattern. This situation should desirably increase the signal above the signal obtained by the same non-operational structure defined in the adjacent track. ^本明明向Image/Positive Embossed Inverted Disc (Processing 丨) or Forward Image: Negative Embossed Inverted Disc (Process 2), the reflective layer 134 is now rendered as the first side of the light. The transparent substrate layer η 2 is presented to assist in laser focusing 'such as a standard optical disc. Figure 6Α Previously explained the problem to the image/positive embossed inverted disc.

在圖6Β』不之-簡單解答為增加—額外聚焦透鏡1 7至光 碟光學頭拾音器。用以產生呈現在例子2和例子3中的資料 之閱讀機在此為如此修正。另外，或是額外，介於光學頭 拾音器和光碟第—矣& β μ n 表面之間的距離可以加以調整以致於雷 射可以正確地聚焦在光碟第一表面上。 另-種替代方案，—較佳具體實施例在下文更進一步說 明，s周整光碟本身而非閱讀機。在此較佳具體實施例中， 非整體或是整體雷射折射構件以蓋板附接至光碟雷射接近 側以建立光碟組件。此蓋板作用為折射，以及因此更進一 98El5.doc -34- 1249158 步將入射光聚焦在光碟操作平面上。傳統上，該操作平面 現在计异為光碟第二表面。雖然目前較佳，但是可安襄敛 練或是另外可修正附接之整體蓋板亦可以使用。 在處理1或是處理2中，標準CD-R鑄模在處理}以標準母 片530沖壓或是在處理2為反向影像父片52〇或是子片54〇_ 產生厚度為1.2 mm之光碟基板。蓋板之加入則將光碟組件 厚度增加為超越制定在標準中之微不足道之丨·2 mm厚度之 • 上。雖然呈現之實驗由此實際說明2.4 mm之深的光碟仍然 可以讀取’但仍然期待將蓋板意圖加入之基板132環境厚 度減低。 在新的方式中，基板132深度可以在處理1或是處理2最 〜v驟中使用DVD-R鑄模而減低。如將欣賞地是，cd-R 格式壓模（處理1為母片530，處理2為反向影像父片52〇或 疋子片54〇)必須製造為可以安裝在鑄模中，而且鑄模設定 • 必須調整以確定適當沖壓。如此形成之基板〇_6 mm ;而具 有丨·2 mm厚之蓋板時，組合光碟組件厚度因此為1.8 mm。 無淪是何種解答或是解答組合為用以調整聚焦，本發明 單一資料層倒置光碟幾何消除基板132所選擇合成物之光 學限制是明顯的，相對於上述透明基板112之光學限制。 也就是說’由於本發明目前單一資料層倒置光碟之層132 不用以折射入射雷射光，和標準光碟基板112相反，層132 透明、折射指數和厚度可以調整而與這些光學參數無關。 隋^/呈現不谷易以標準光碟達成的製造優點。然而，既 疋現存光碟製造裝置之安裝基座，目前以使用聚碳酸脂製 988l5.doc -35- 1249158 造本發明單一資料層光碟為較佳，如下文說明。 為實際說明這些原理可用以建造具有同時可讀取非操作 結構之可尋軌光碟，單一資料層前向影像/正浮雕倒=光 碟如例子1所詳細陳述加以製造。各光碟基板132由注射 鑄造之聚礙酸脂組成’包含具有L6 _節距之擺動紋道前 向影像以及擺動頻率近似在CD_R標準中陳述之頻率。金 層配置在雷射接近表面以形成反射層13 4。In Figure 6 Β No - simple solution for adding - extra focus lens 1 7 to the optical pickup of the optical disc. The reader used to generate the data presented in Examples 2 and 3 is modified as such. Alternatively, or additionally, the distance between the optical pickup and the surface of the disc - 矣 & β μ n can be adjusted so that the laser can be properly focused on the first surface of the disc. Another alternative, the preferred embodiment, is further described below, in which the entire disc itself is not a reader. In the preferred embodiment, the non-unitary or unitary laser refractive member is attached to the proximity of the disc laser with a cover to establish the optical disc assembly. This cover acts as a refraction, and thus a step 98E5.doc -34-1249158 focuses the incident light on the disc operating plane. Traditionally, this plane of operation is now counted as the second surface of the disc. Although currently preferred, it is also possible to use an integrated cover that can be spliced or otherwise modified. In Process 1 or Process 2, the standard CD-R mold is processed by the standard master 530 or the process 2 is the reverse image parent 52 〇 or the sub-piece 54 〇 _ produces a disc of 1.2 mm thickness Substrate. The addition of the cover increases the thickness of the disc assembly beyond the negligible thickness of 2 mm set in the standard. Although the experiment presented thus actually shows that a disc of 2.4 mm depth can still be read', it is still expected that the environmental thickness of the substrate 132 to which the cover is intended to be added is reduced. In the new approach, the depth of the substrate 132 can be reduced by using the DVD-R mold in either Process 1 or Process 2. As will be appreciated, the cd-R format stamper (process 1 is master 530, process 2 is reverse image master 52 〇 or tweezer 54 〇) must be made to fit in the mold, and the mold is set • Must be adjusted to determine proper stamping. The substrate thus formed is 〇6 mm; and when the cover is 丨·2 mm thick, the thickness of the combined optical disk assembly is therefore 1.8 mm. What is the solution or combination of solutions for adjusting the focus, the optical limitations of the selected composition of the single data layer inverted optical disc geometry eliminating substrate 132 of the present invention are significant relative to the optical limitations of the transparent substrate 112 described above. That is, since the layer 132 of the present single-data layer inverted optical disk is not used to refract incident laser light, as opposed to the standard optical disk substrate 112, the transparency, refractive index and thickness of the layer 132 can be adjusted independently of these optical parameters.隋^/ presents the manufacturing advantages that are not achieved with standard optical discs. However, in addition to the mounting base of the existing optical disc manufacturing apparatus, it is preferable to fabricate the single data layer optical disc of the present invention using polycarbonate 988l5.doc -35-1249158, as explained below. To illustrate these principles, it is possible to construct a track-receivable optical disc having a simultaneously readable non-operating structure, and a single data layer forward image/positive embossing = optical disc is manufactured as detailed in Example 1. Each of the optical disk substrates 132 is composed of injection-molded agglomerated acid grease. The forward image of the wobble track having an L6_pitch and the wobble frequency are approximately the frequencies stated in the CD_R standard. The gold layer is disposed near the surface of the laser to form a reflective layer 132.

在第一系列實驗中，在例子2中詳細報告，特定於人體 免疫寫球素G(IgG)之驗定格點建構在光碟小部分氣體入射 金表面上。 如圖7A所示意，該驗定格點建構為三層爽心；將由那些 熟知臨床驗定相關技藝之人士所瞭解地是，該夾心目的為 呈現最終夾心層，抗體76為用於分析物捕獲和.偵測。將由 那些熟知相關技藝之人士所瞭解地是，該驗定本身包含許 多此種三分子夾心，只有其中之一為在圖7中示意。 在第一系列實驗中，抗體76因為其特定性而選擇用於人 體免疫血球素G(IgG)，在正常人體血液近似於丨」g/dl之濃 度中發現。 由成人志願者所抽出之人體血液樣本被應用於光碟驗定 格點以及簡易地在上面培養。該光碟被清洗，而且之後則 由3 μπι膠乳球體之應用而加以”顯影”，各球體先前已經以 人體IgG特定之抗體加以塗層。該光碟之後再次被清洗。 如圖7B所示意，此處理之結果為膠乳球體乃之“①調解特 定附著於光碟。在血液樣本中缺乏IgG時，該膠乳球體將 988l5.doc -36- 1249158 在清洗期間被移除。 圖8為由光線微觀檢視部分IgG特定光碟捕獲之視頻影 像，㈣乳球«接著人體血液和抗體_共輛膠乳球體。 該膠乳球體為容易出現，如擺動紋道本身。在此視頻捕獲 影像中，較暗區域為正尋執結構。放大率預先排除正觀察 ”紋道，，連續性。該視頻捕獲影像建議假使不是全部但也是 井多膠乳球體位於鄰近在此前向/正浮雕光碟上的擺動紋 道。 必須容易出現地是相對於紋道寬度之發出信？虎一部份大 小可以有利地加以調整而加強非操作結構之定位。例子4 :現發出信號-部份大小和紋道寬度之最佳化關係的計 算，在環境中，前向影像/負浮雕倒置光碟被使用。類似 地是，在驗定袼點上的分子範圍之大小，此處三分子維生 素H-Streptavidin-維生素η夾心可以有利地加以調整而加強 此，出信號一部份的穩健、然而限制移動而允許該定位。 證實有利於發出信號一部份的變化長度之聚合脊骨在共同 擁有以及上在審理中之美國專利申請案序號為第 09/12Μ49號，1998年7月21日中請，·第〇8細，935號， 1997年7月7日中晴中說明；該文獻在此處以提及的方式併 入本文中。甚至於，紋道本身形狀在CD和DVD規範中可 以調整以加強該定位。 圖9和圖1〇為附著於光碟之單一膠乳球體的原子力 (ATM)微觀影像，為稍高於圖8中使用的放大率。在圖9為 谷易顯著的是該擺上紋道本身以及膠乳球體定位於接近紋 98815.doc -37- 1249158 道-轉之中央。圖10呈現編音取之量化資料。該紋道 量測171.70 nm深。㈣球體超過紋道底部的高度為24〇7 μηι 〇 圖11為二膠乳球體在單一微觀視界中之原子力微觀影 像。該量化區段分析解釋用以顯影此抗體_IgG免疫驗定之 膠乳球體的-致性。另外，該量化區段分析辨識介於紋面 中心和鄰近紋道中心之間的水平距離為843 75 nm(0 84 μηι) ’和所需之1 ·6 μιη磁執為優良地吻合。 在例子3中報告之第二系列實驗，實際說明此種非操作 結構可以可靠地由最小修正CD_R裝置偵測為hf信號高 頻、高振幅變化。也就是說，上述具有擺動紋道之單一資 料層、前向影像/正浮雕第一表面驗定光碟允許尋軌信號 和配置在光碟表面上的非操作結構產生之信號同時擷取以 及由該信號鑑別。 如例子3更進一步說明，光碟如例子丨所陳述地準備。驗 定袼點基本上如例子2準備，但是以抗_glyc〇ph〇rin抗體取 代在例子2所使用之抗-lgG抗體76。Glycophorin為出現在 所有人體紅血球（紅血球細胞，RBC)表面上的蛋白質。 人體血液之肝燐脂化樣本（1〇…丨應用至驗定格點，以及 該光碟之後簡易地洗濯。 圖12為確定RBc免疫特定附著於光碟驗定格點的原子力 微觀影像。如量化分析所表示，RBC水平大小既定為7.984 ’和紅血球細胞（8 μηι)已知直徑吻合；此大小和例子2 使用與觀察之膠乳球體均勻的3 μηι直徑為明顯地不同。 98815.doc -38- 1249158 RBC超過紋道底部的高度觀察為18 μηι。 如例子3更進一步說明，由CD Associates，Inc.(加州 Irvine市）所製造用於光碟工業品質控制使用的cd-R裝置為 用以讀取該光碟。驅動裝置的CD-R擺動尋執系統（模式 RSL100)被加入之透鏡17修正光學拾音器以在光碟上缺乏 折射層112調整聚焦；轉軸之高度亦提高。hf信號（亦mRF 或是四列總和）由RSL 100中的電子電路加以放大，以致於 不波器顯示可以在不用不利於影響擺動尋執裝置效能下提 供。圖1 3呈現代表性尋執，以X軸顯示時間而γ軸顯示四 列總和信號大小。 圖13實際說明紅血球細胞直接可見地為cD-R閱讀機hf 4吕號面頻、南振幅事件；對哺乳動物細胞大小之分析物而 言，沒有膠乳球體或是其他外因性發出信號一部份為所需 以產生特定分析物信號。 亦由圖13之示波器追蹤明顯地是hf基準線的差異為雙 重峰值。雖然紅血球細胞已知為具有特徵性雙凹形狀，我 們已經在例子2中觀察到此雙重峰值被用以報告分析物之 呈現。當雷射遍歷球體時，此雙重峰值出現為由反射比中 之再生變化形成。此種再生電子記號可以有利地用以由各 種不同大小的非操作結構辨識以及鑑別信號。 由圖13之示波器追蹤另一觀察而言，容易見到地是在發 出信號事件各側上的基準線為穩定；也就是說，擺動紋道 之尋軌本身不造成四列總和信號中的重大變化。 如CD-R標準之光學閱讀機，無論正在光碟上的任何位 98815.doc -39- 1249158 置讀取都維持常數線性速度，修正轉軸速度以鎖定擺動紋 道之頻率。依據已知光碟線性速度和示波器尋軌上的增加 標示，示波器追蹤上的各劃分可以顯示為相當於光碟：線 性距離13 μπι。如圖13顯示之示波器尋軌所量測，四列總 和信號基準線中的差異因此既定為1G叫，近似尋軌㈣ 方向之物件的非校正大小。 該物件實際大小為較小。使用3 μηι膠乳球體之閱讀機校 準和不波器已經給予示波器峰值報告5 μηι明顯大小，較實 際物件寬2 μιη。此情形適當地在驗定光碟上㈣一表面上 考量為1·5 μηι雷射聚焦直徑。 考量介於量測和雷射直徑在光碟表面上發生實際大小之 間的2㈣差異時’在圖13示波器尋執捕獲的事件為四列總 和信號高頻、高振幅差異’報告8叫物件大小，和已知8 μιη人體紅血球吻合。 圖14呈現由相同光碟上之分離紅血球細胞侦測發出信號 的㈣件之另—示波器追蹤。雙相峰值為更顯著。圖15_ 17為額外例子。 β _由相同光碟上不同區域掏取之多數事件的數位重 逢實際5兒明在-些特定免疫附著於光碟不同紅灰球細胞 上大小和形狀量測之再生。 總結’例子i-3實際說明配置在如由此說明之原裡建構 的單-層倒置光碟第一表面上微米大小之非操作結構，可 以由最小修正之標準光碟讀寫器加以偵測、量測和特徵 化。包含尋軌結構之光碟操作結構為使用單—光學拾音考 98815.doc -40- 1249158 门日守與非操作結構同時積測月 才1貝成1以及谷易由非操作結構鑑別。 例子2特別實際說明小分子分蚯 . 刀于刀析物之免疫分析可以容易地 _適為使用此系統偵測；例早 』例子3特別實際說明細胞計數和 細胞分析亦容易地完成。 雜化作用之驗定可以類似 加以彳貞測。 例子7依次實際說明依據核酸 地調適為使用本發明可尋軌光碟In the first series of experiments, it was reported in detail in Example 2 that the assay site specific to human immunoglobulin G (IgG) was constructed on a small portion of the gas incident gold surface of the disc. As illustrated in Figure 7A, the assay grid is constructed as a three-layered refreshment; it will be appreciated by those skilled in the art of clinical assays that the sandwich is intended to present the final sandwich layer and antibody 76 is used for analyte capture and. Detection. It will be appreciated by those skilled in the art that the assay itself contains many such three-molecule sandwiches, only one of which is illustrated in Figure 7. In the first series of experiments, antibody 76 was selected for human immunoglobulin G (IgG) because of its specificity, and was found in normal human blood at a concentration similar to 丨"g/dl. Human blood samples taken from adult volunteers are applied to the disc verification grid and simply cultured on it. The disc is cleaned and then "developed" by the application of 3 μm latex spheres, each of which has previously been coated with human IgG specific antibodies. The disc is cleaned again afterwards. As shown in Fig. 7B, the result of this treatment is that the latex sphere is "1 mediating specific attachment to the disc. When the blood sample is deficient in IgG, the latex sphere is removed during the cleaning period of 988l5.doc -36-1249158. 8 is a video image captured by a light microscopic view of an IgG-specific disc, (4) a milk ball «and a human blood and an antibody _ a total of latex spheres. The latex sphere is prone to occur, such as the wobble track itself. In this video capture image, The darker area is the positive search structure. The magnification is precluded from observing the "track", continuity. The video capture image suggests that not all but also the multi-lact spheres are located adjacent to the oscillating tracks on the omni/positive embossed disc. Must be easy to appear is a letter relative to the width of the groove? A part of the tiger can be advantageously adjusted to enhance the positioning of the non-operational structure. Example 4: The calculation of the signal-partial size and the width of the groove width is calculated. In the environment, the forward image/negative embossed inverted disc is used. Similarly, in determining the size of the molecular range on the defect, here the three-molecule vitamin H-Streptavidin-vitamin η sandwich can be advantageously adjusted to enhance this, allowing a portion of the signal to be robust, yet restricting movement to allow The positioning. U.S. Patent Application Serial No. 09/12,49, the entire disclosure of which is incorporated herein by reference in its entirety in its entirety in its entirety in , 935, July 7, 1997, the disclosure of which is incorporated herein by reference. Even the shape of the track itself can be adjusted in the CD and DVD specifications to enhance the positioning. Figures 9 and 1 show the atomic force (ATM) microscopic image of a single latex sphere attached to a disc, which is slightly higher than the magnification used in Figure 8. In Fig. 9, it is obvious that Gu Yixian is that the pendulum itself and the latex sphere are positioned in the center of the pattern - 98815.doc -37-1249158. Figure 10 shows the quantitative data obtained by the timbre. The groove measures 171.70 nm deep. (4) The height of the sphere beyond the bottom of the groove is 24〇7 μηι 〇 Figure 11 is the atomic force microscopic image of the second latex sphere in a single microscopic horizon. This quantitative section analysis explains the susceptibility of the latex spheres used to develop this antibody-IgG immunoassay. In addition, the quantized segment analysis identified that the horizontal distance between the center of the land and the center of the adjacent groove was 843 75 nm (0 84 μηι) ' and the required magnetic resonance of 1 · 6 μιη was excellently matched. The second series of experiments reported in Example 3 demonstrates that such a non-operational structure can be reliably detected by the minimally modified CD_R device as a high frequency, high amplitude variation of the hf signal. That is, the single data layer having the wobble track and the forward image/positive relief first surface verification optical disc allow the tracking signal and the signal generated by the non-operational structure disposed on the surface of the optical disk to simultaneously capture and be used by the signal. Identification. As further illustrated in Example 3, the optical disc is prepared as set forth in the example. The assay sites were prepared essentially as in Example 2, but the anti-lgG antibody 76 used in Example 2 was replaced with an anti-glyc〇ph〇rin antibody. Glycophorin is a protein that appears on the surface of all human red blood cells (red blood cells, RBC). The liver lipids of human blood are sampled (1〇...丨 applied to the assay grid, and the disc is simply washed after the disc. Figure 12 is a microscopic image of the atomic force that determines the specific attachment of the RBc to the disc. As quantized The RBC level was determined to be 7.984 ' and the red blood cells (8 μηι) were known to be diameter-matched; this size and Example 2 were significantly different from the observed 3 μηι diameter of the latex sphere observed. 98815.doc -38- 1249158 RBC exceeded The height of the bottom of the track was observed to be 18 μη. As further illustrated in Example 3, the cd-R device manufactured by CD Associates, Inc. (Irvine, Calif.) for use in optical disc quality control was used to read the disc. The CD-R wobble seek system of the drive unit (mode RSL100) is added to the lens 17 to correct the optical pickup to adjust the focus on the lack of the refractive layer 112 on the disc; the height of the shaft is also increased. The hf signal (also mRF or the sum of the four columns) It is amplified by the electronic circuit in the RSL 100, so that the non-wave device display can be provided without adversely affecting the performance of the swing seek device. Figure 13 shows the representative The search is performed on the X-axis and the γ-axis shows the four-column sum signal size. Figure 13 shows that the red blood cells are directly visible to the cD-R reader hf 4 Lu, the south amplitude event; For analytes, there is no latex sphere or other exogenous signaling that is required to produce a specific analyte signal. The oscilloscope trace of Figure 13 is clearly the difference in the hf baseline as a double peak. Although red blood cells Known as having a characteristic biconcave shape, we have observed in this example that this double peak is used to report the presentation of the analyte. When the laser traverses the sphere, this double peak appears as a regenerative change in the reflectance. Such regenerative electronic signatures can advantageously be used to identify and identify signals from a variety of different sized non-operational structures. From the oscilloscope trace of Figure 13, another observation is readily seen as a reference on each side of the signalling event. The line is stable; that is, the tracking of the wobble track itself does not cause significant changes in the four-column sum signal. Optical readers such as the CD-R standard Regardless of the position of the 98815.doc -39- 1249158 on the disc, the constant linear velocity is maintained, and the spindle speed is corrected to lock the frequency of the wobble track. According to the known linear speed of the disc and the increased indication on the oscilloscope tracking, Each division on the oscilloscope trace can be displayed as equivalent to a disc: linear distance 13 μπι. As shown in the oscilloscope tracking shown in Figure 13, the difference in the four-column sum signal reference line is therefore set to 1G, approximate tracking (four) direction The non-corrected size of the object. The actual size of the object is small. The calibrator and the non-wave device using the 3 μηι latex sphere have given the oscilloscope a peak report of 5 μηι, which is 2 μηη wider than the actual object. This situation is appropriately considered to be a laser focus diameter of 1·5 μηι on the (four) surface of the verification optical disc. Consider the difference between the measurement and the actual diameter of the laser diameter on the surface of the disc. 'The event captured by the oscilloscope in Figure 13 is the four-column sum signal high frequency, high amplitude difference' report 8 called object size, It is consistent with the known 8 μιη human red blood cells. Figure 14 presents an additional oscilloscope trace of (4) detected by the isolated red blood cells on the same disc. The two-phase peak is more pronounced. Figure 15_17 is an additional example. β _ The number of most events taken from different areas on the same disc reproduces the actual size and shape of the specific nucleus attached to different red ash cells on the disc. Summary 'Example i-3 actually illustrates the micro-sized non-operating structure configured on the first surface of a single-layer inverted optical disc constructed as described above, which can be detected and quantified by a minimally modified standard optical disc reader. Measure and characterize. The optical disc operating structure including the tracking structure is to use the single-optical pickup test 98815.doc -40-1249158. The gate and the non-operational structure simultaneously measure the month, and the 1st and 1st are identified by the non-operational structure. Example 2 specifically illustrates the small molecule bifurcation. The immunoassay of the knife in the cleavage of the cleavage can be easily adapted to the use of this system; the early example of Example 3 is particularly practical to demonstrate that cell counting and cell analysis are also easily accomplished. The verification of hybridization can be similarly speculated. Example 7 sequentially illustrates the adaptation of the nucleic acid to the use of the present invention.

士如例子7所詳細陳述’溶解相位雜化作用為在下列常數 什數呈現之目標核酸各種不同濃度平行執行：⑴順序至第 :部分目標（，，3,探針”）互補單—股核酸探針和⑺順序至第 刀目標（"5'楝針”）互補單—股核酸探針。該3，探針為更 進—步共軛至順磁性串珠以及該5,探針為更進一步共軛至 蛋白質：順磁性串珠作用為允許磁性分離以及部分雙工目 標之淨化’以及更進一步允許由光碟閱讀機直接偵測；5, 探針蛋白質一部份允許由應用至本發明光碟第一表面之金 屬表面的streptavidin捕獲部分雙工目標。 圖36示意在視覺化時間之驗定格點。直接附著於可尋執 光碟金表面為由韋德瓦斯力以及介於自由sulfyhydryi streptavidin蛋白質與光碟金表面之間形成的硫化_金黏合 劑接合之streptavidin鍍層。該streptavidin捕獲5，探針之維 生素Η—部份。該5,探針依次由14核甘酸之Wats〇n_Crick補 充在目標3’端捕獲目標核順序。該目標依次在5,端經由14 核甘酸之Watson-Crick補充捕獲3,端，因此繫鏈Dynabead® 至該光碟。 圖37呈現分別採取之驗定光碟ι-3光微觀影像，各光碟 98815.doc -41 - 1249158 放大率為2:附接球體和擺動紋道在所有3光碟較高放大率 面板上為清楚可見。增加數目之附接串珠以增加量之核酸 目標為清楚可見，而光碟3(圖37C)顯示補充驅動之球體以 20十兆億分之一莫耳（2〇χ1〇-2ΐ莫耳；12χ1〇3分子）核酸目標 附接至光碟表面，而光碟2(圖37Β)顯示補充驅動之球體以 20百億億分之一莫耳（2〇xl〇w莫耳；12χ1〇ό分子）核酸目標 附接至光碟表面，以及光碟丨（圖37Α)顯示補充驅動之球體 以20千萬億分之一莫耳（2〇xl(rl5莫耳；12χ1〇9分子）核酸目 標附接至光碟表面。各控制光碟面上沒有觀察到任何串珠 (沒有顯示）。將欣賞地是圖37在各影像中只呈現一部份驗 定範疇。 例子7因此實際說明核酸雜化作用基準之驗定可以容易 地調適為使用本發明光碟加以偵測。在可以容易地調適為 使用本發明光碟加以偵測之核酸雜化作用基準的驗定間為 雜化作用至核酸陣列，如美國專利案號為第5,143,854號； 5,405,783 號；5,445,934 號；5,510,270 號；5,800,922 號； 5,556,752 號；5,599,695 號；5,744,305 號；5,770,722 號； 5,837,832號；5,843,655號；國際專利出版案號為w〇 98/12559(DmerS)中加以說明，以及在 Nature Genetics 21 (Suppl·) : 1-60( 1999)更進一步說明，該文獻在此處以提 及的方式併入本文中。 例子7更進一步實際說明磁性串珠，長期使用於分子生 物學之分離和淨化，現在將另外直接使用於發出信號，提 供分離、淨化和偵測之有效系統而不強制以物理放射性核 98815.doc -42- 1249158 種、fluorophores、酵、化學一部份或是類似物另外標示 核酸。 共同擁有以及上在審理中的申請案序號第〇8/888,935 號，1997年7月7曰申請；第〇9/120,049號，1998年7月21曰 申請在此處以提及的方式併入本文中，說明各種其他信號 元素和化學將允許現存驗定調適為使用本發明可尋軌光碟 之偵測。 在先前提到之各利用中，本發明光碟允許獨立、立體可 分解事件。 單一資料層光碟變化 將由熟知相關技藝之人士所瞭解地是，單一資料層特定 分析物光碟之許多變化130，可以加以製造。不限制之多 數具體實施例將在下文討論。 全像操作結構 現在參考圖20，顯示單一資料層具體實施例。在此具體 貫施例中光碟之操作結構編碼在反射全像而非藉由實體 壓製在光碟基板中。反射全像允許操作結構之實體平面由 非操作結構佔據的實體平面移除。多數資料層（資訊層）光 碟’如下文更進—步說明，纟現另—方式於操作結構和非 私作結構之分離。 光碟190包括光碟基板192、全像194和透明防護鍍層 198。全像194為包含光碟19〇所需之操作結構的反射全 像。特定地是，當雷射由全像194反射時，假使正確方位 之擺動紋道呈現在全像影像平面195上時將出現。 98815.doc -43- 1249158 較佳具體貫施例中，全像影像平面195雷射接近至 王像實體平面194以及實質上與配置在驗定光碟第一表面 j非操作結構！ 96共焦。該雷射如之前被聚焦在由非操 乍、Ό構和操作結構（此處，擺動紋道影像）共享的平面上， 允许細作資料（特定為尋軌資料）和特定分析物資料。 當然，應該為明顯地是將雷射最佳化聚焦在全像影像平 —非操作結構共焦_需要雷射較不嚴謹地聚焦在全像 _平面上。而全像非常自然地是不僅容許該”誤差”亦且 由此得利。如已在光學技藝所知，各部分全像實體表面可 以產生影像之完整性，干涉地編碼在該表面上，然而如照 射部分大小減少，解析度減低。相反地，全像較大部分照 射時，影像較佳。因此，照射雷射點較大，光碟操作結影 像較佳-在較佳具體實施例中，將為_擺動紋道。 將由熟知相關技藝之人士所瞭解地是，全像影像平面亦 可以有利地投射，所以不再正確地與非操作結構共焦，至 於非操作結構如擺動紋道為與特定分析物信號為同時可偵 測。因此，該影像不僅可以雷射接近至全像時平面，亦且 為雷射遠端。 甚至於，雖然顯示為整合至光碟190，全像1 94為可移 除。此情形允許全像194將利用現存高速全像印製處理加 以大量生產。甚·至於，和應用有關，全像194亦可以相反 地附接於光碟基板丨92，潛在地允許基板丨92再使用。本發 明可尋軌驗定光碟之全像單一資料層具體實施例允許由光 碟閱讀機廣泛安裝機座可讀取之光碟為低成本大量生產。 98815.doc -44- 1249158 另一方面’本發明單一資料層光碟實體浮凸壓印之具體 實施例中，光碟基板192不需要符合標準透明光碟基板112 之光學需求，因為光碟基板1 92位於資料平面之雷射遠 端。 亦且如上述之具體實施例中，其中該操作結構分別被浮 凸壓印和反射鍍層，全像具體實施例經常可以有利地包含 輔助I焦之雷射接近、非整合蓋板。在全像具體實施例 中，焦點因此可以調整在全像影像平面上。As detailed in Example 7, the 'dissolved phase hybridization' is performed in parallel at various concentrations of the target nucleic acid present in the following constants: (1) sequence to: part of the target (, 3, probe)) complementary single-stranded nucleic acid The probe and (7) sequence to the cleavage target ("5' 楝") complementary single-strand nucleic acid probe. The 3, the probe is more forward-conjugated to the paramagnetic bead and the 5, the probe is further conjugated to the protein: paramagnetic beading to allow magnetic separation and purification of part of the duplex target' and further allow Directly detected by a disc reader; 5, a portion of the probe protein allows for capture of a portion of the duplex target by streptavidin applied to the metal surface of the first surface of the disc of the present invention. Figure 36 illustrates the checkpoint of the visual time. Directly attached to the gold surface of the rewritable disc is a streptavidin coating bonded by a vadovas force and a vulcanized-gold bond formed between the free sulfyhydryi streptavidin protein and the surface of the disc gold. The streptavidin captures 5, the vitamin Η part of the probe. In the 5th, the probe is sequentially supplemented with Wats〇n_Crick of 14-nuclear acid to capture the target nuclear sequence at the target 3' end. The target was then replenished at the 5' end with a 14-nucleotide Watson-Crick to capture the 3' end, thus tethering Dynabead® to the disc. Figure 37 presents the optical micrographs of the optical discs ι-3 taken separately, each disc 98815.doc -41 - 1249158 Magnification 2: Attached spheres and oscillating trajectories are clearly visible on all 3 discs higher magnification panels . The increased number of attached beads is clearly visible to increase the amount of nucleic acid target, while the disc 3 (Fig. 37C) shows the complementary driven sphere at 20 billion trillion moles (2〇χ1〇-2ΐ莫耳; 12χ1〇) The 3 molecule) nucleic acid target is attached to the surface of the disc, and the disc 2 (Fig. 37A) shows the complementary driving sphere with a nucleic acid target of 20 billionth of a mole (2〇xl〇wmole; 12χ1〇ό molecule) Connected to the surface of the disc, and the disc 丨 (Fig. 37Α) shows that the complementary drive sphere is attached to the surface of the disc with a 20 mega-mole (2 〇 xl (rl 5 mr; 12 χ 1 〇 9 molecule) nucleic acid target. No beading is observed on the control disc surface (not shown). Appreciation is shown in Figure 37. Only a part of the verification range is presented in each image. Example 7 Therefore, the verification of the nucleic acid hybridization benchmark can be easily adjusted. For detection using the optical disc of the present invention, hybridization between the assays that can be readily adapted to the nucleic acid hybridization criteria detected using the optical disc of the present invention is hybridization to a nucleic acid array, such as U.S. Patent No. 5,143,854; 5,405,783; 5,445,9 No. 34; 5, 510, 270; 5,800, 922; 5,556,752; 5,599,695; 5,744,305; 5,770,722; 5,837,832; 5,843,655; International Patent Publication No. (Suppl.): 1-60 (1999) further clarifies that this document is incorporated herein by reference. Example 7 further illustrates magnetic beading, long-term use in the separation and purification of molecular biology, now It will additionally be used directly to signal, providing an efficient system for separation, purification, and detection without forcing additional identification of nucleic acids with physical radionuclides 98815.doc -42-1249158, fluorophores, leavens, chemical moieties, or the like. Co-ownership and the pending application number No. 8/888,935, July 7, 1997, application; No. 9/120,049, July 21, 1998, the application is hereby incorporated by reference. It is stated that various other signal elements and chemistries will allow the existing assay to be adapted to detect the use of the track-receivable optical disc of the present invention. In each of the previously mentioned uses, the present invention The disc allows for independent, stereoscopically decomposable events. A single data layer disc change will be known to those skilled in the art, and a number of variations of a single data layer specific analyte disc can be made. Most embodiments will be The following is a discussion of a holographic operation structure Referring now to Figure 20, a single data layer embodiment is shown. In this particular embodiment, the operational structure of the optical disc is encoded in a reflective holographic image rather than being physically pressed into the optical disc substrate. The reflection hologram allows the physical plane of the operational structure to be removed by the physical plane occupied by the non-operational structure. Most data layer (information layer) discs are further described in the following paragraphs, and the other way is to separate the operational structure from the non-private structure. The optical disk 190 includes a disk substrate 192, a full image 194, and a transparent protective plating 198. The hologram 194 is a reflection hologram of the operational structure required to contain the optical disk 19 。. In particular, when the laser is reflected by the hologram 194, it will occur if the correct orientation of the wobble track appears on the hologram image plane 195. 98815.doc -43- 1249158 Preferably, in the embodiment, the holographic image plane 195 is close to the king image solid plane 194 and substantially non-operating with the first surface of the optical disc. 96 confocal. The laser, as previously focused on a plane shared by non-operating, framing, and operational structures (here, oscillating striated images), allows fine-grained data (specifically tracking data) and specific analyte data. Of course, it should be obvious that the laser is optimized to focus on the holographic image - the non-operating structure confocal _ requires that the laser focus less on the holographic _ plane. The hologram is very natural to not only allow this "error" but also benefit from it. As is known in the art of optics, the holographic solid surface of each portion can produce image integrity and be interferometrically encoded on the surface, however, as the size of the illuminating portion is reduced, the resolution is reduced. Conversely, when the hologram is illuminated for a larger portion, the image is better. Therefore, the illumination laser spot is large, and the disc operation image is preferably - in the preferred embodiment, the _ wobble track. It will be appreciated by those skilled in the relevant art that the holographic image plane can also be advantageously projected, so that it is no longer correctly confocal with the non-operating structure, as for non-operational structures such as wobbled tracks that are simultaneously with a particular analyte signal. Detection. Therefore, the image can not only be close to the plane of the full image, but also the far end of the laser. Even though, although shown to be integrated into the disc 190, the hologram 1 94 is removable. This situation allows the hologram 194 to be mass produced using existing high speed holographic printing processes. As a matter of application, the full image 194 can also be attached to the optical disk substrate 丨 92, potentially allowing the substrate 丨 92 to be reused. The present invention can be used to determine the holographic image of a compact disc. A single data layer embodiment allows a wide range of optical discs that can be read by a disc reader to be mass produced at low cost. 98815.doc -44- 1249158 In another embodiment of the present invention, the optical substrate 192 does not need to conform to the optical requirements of the standard transparent optical disk substrate 112 because the optical disk substrate 1 92 is located in the data. The laser's distal end of the plane. Also in the specific embodiment described above, wherein the operational structure is embossed and reflective coated, respectively, the holographic embodiment may often advantageously include a laser-assisted, non-integrated cover of the auxiliary I-focus. In the holographic embodiment, the focus can therefore be adjusted on the holographic image plane.

區域化CLV 由上文說明之實體壓印和全像編碼之單一資料層光碟， 二者經常可以利用和CD_R標準擺動紋道不同的尋軌設 e十。特別疋，本發明光碟可以如本發明以位於光碟區域外 之（ZCLV)格式原理”Zoned Constant Unear Vel〇city„加以建 立。此處簡易地加以說明，（ZCLV)格式如相關技藝所熟 知，在各種不同工業標準詳細化，包含dvd_ram規範（例 如請參考ECMA 272，此處以提及的方式併入本文中）。 如上側視圖21所顯示，ZCLV光碟2〇〇具有資料區2〇2， 劃分為多數區域204a-204e。雖然只有顯示5個區域，實際 ZCLV格式光碟可以具有不同數目之區域。例如，dvd_ RAM ZCLV袼式具有24個區域在可寫人㈣區域内。 各區域2〇4a_204e劃分為多數扇區2〇6。内部區域具有較 外部區域少的扇區，因為内部區域半徑小於外部區域半 徑。光碟之佈局被配置為各磁執（亦即螺旋各轉）上的資料 塊槽頭資訊在各扇區内為徑向對齊。此情形允許將用於各 988l5.doc -45- 1249158 ::==，_區域緊接著資料 在使用日夺，光碟閱讀機可以各區域内 光碟以及仍然維㈣域㈣常數資料、對^ =言以維_上整體_ 圏2”s 域而…亥光碟可以較低速率轉動。 圖22顯不ZCLV光碟2〇〇其中之一扇區的一部份 圖。如所能看見，多數磁軌220在扇區内為徑向配置，所 ==在”預製紋道，，區域222浮凸壓印之資料塊的 田貝貝訊。各磁執資料之後可以在擺動紋道内和.，擺動紋 :和紋道”區域224内之紋道間的擺動紋面區域上加以記 為如本發明使用為驗定光碟，非操作結構可以配置 在光碟上之扇區之擺動紋面和紋道區域224内，如圖U所 顯不。因為在各扇區内，擺動紋面和紋道區域“A形成連 績區域’使用ZCLV格式之驗定光碟可以使用各扇區執行 不同驗疋在預製紋道區域222中的浮凸壓印檔頭資訊可 用以儲存辨識擺動紋面和紋道區域内之驗定的資訊。該擺 動紋面和紋道區域内之擺動紋面和紋道以及在預製紋道區 域中的洋凸壓印資料磁執滿足光碟閱讀機之功能需求。 如上文所建議，各種不同數位多樣化光碟（DVD)標準之 特徵可以有利地在本發明單一資料層具體實施例中之實現 加以利用。所以，本發明不限於現存CD_類型光碟或是cd 標準。除了 ZCLV格式特定利用於上文討論之多數驗定光 98815.doc -46- 1249158 碟以外’已經明顯地是制訂於DVD標準之較小結構大小和 降低波長雷射允許較高密度之非操作結構將以較可能使用 CD標準為高的立體鐘別。調適dvd_ram標準之態樣為本 發明強向影像倒置光碟的另—重大優點為其紋面-紋道尋 軌態樣（參考職A 272)，將可用於Μ上之非操作結構横 測和特徵化的切線解析度加倍。甚至於，提供獨特特徵之 雙層資料層DVD格式將在信文章節中更進一步討論。 光碟蓋板 由此使用於例子2和例子3中的CD_R閱讀機在此已經藉 由增加聚焦透鏡17而修正為考量缺乏特定分析物光碟第一 表面之雷射接近折射基板時調整驅動裝置焦點。經常將優 先採用之另一方式為調整光別本身而非光碟閱讀機。在之 後的方式中’雷射折射構件為附接於光碟雷射接近側而為 蓋板，该蓋板作用為折射以及因此聚焦該入射光在光碟折 射表面上。合適地設計時，該蓋板在驅動裝置聚焦光學中 排除另一交替。使用此種蓋板而獲得呈現在圖4〇中的資 料’而不用另一聚焦透鏡17之輔助。 此種方式描繪在圖19中。在此藉由傳統方式，雷射光由 此側面橫斷面圖底部入射。光碟130包括光碟基板132和反 射層13 4 ’非操作結構13 6配置在反射層上。擺動紋道13 8 在基板132中壓製以及由反射層134加以鍍層，如指示。亦 且顯示在圖中的是非整合蓋板140。此種蓋板之一具體實 施例更進一步在圖35中以上透視圖更進一步顯示。 較佳地是，光碟組件（光碟加上附接蓋板）以近似於一元 98815.doc -47- 1249158 化光碟之大小標準加以定大小，也就是說，深度為1 2 mm 以及直徑為80 mm或是120 mm。然而，亦預期地是該光碟 組件可以由此大小加以變化。在之後這些案例中，不論光 碟標準之實體大小規範之變化，該組件必須仍然證實能夠 符合驅動裝置之必要光學和機械需求··在這些需求間，該 雷射必須正確地聚焦在光碟操作平面上，該光碟組件必須 適當地央住轉軸，以及該光碟組件必須和標準重量變化不 大以致於驅動裝置馬達不能維持適當的轉動速度。 在此呈現之例子6在下文說明聚碳酸脂製造之蓋板近似 M7 mm厚，但是除此以外大小相當於標準的i2〇 光 碟。該蓋板以聚碳酸脂製造為利用聚碳酸脂的熟知光學品 質以及利用裝置調適於其鑄造之現成使用性。然而，如下 文更進一步討論，其他塑膠可以有利地使用於建造光碟蓋 板。 如例子5製造之單一資料層光碟為12+/_〇〇5麵厚；如 例子6製造之單一資料層光碟為大約丨^厚。由紅皮書標 準(所有組合層為^ _ 15 mm)提供之實體厚度外部組合為 二蓋板之深度近似2·4随。雖然此情形不會呈現光學問 題-保持瘥規範内的光學路徑_看似可能為此增加厚度可能 見夾住驅動4置中之光碟組件的困難度。甚至於，該組 合之光碟和蓋板超越標準、—元化光碟重量。 然而’實驗性地我們不期望地發現箝夾和轉動之組件沒 有二題i現在圖4〇之資料為使用如例子5製造之光碟而 I侍以及在光碟機讀取之前以依據例子6製造之蓋板組 98815.doc 1249158 合。該蓋板提供足夠的輔助於聚焦以排除另一聚焦透鏡之 增加至光碟機拾音器於產生在圖40陳述之資料。 在一系列分離之實驗中（沒有顯示），我們在附接蓋板之 刖由光碟移除箝夾環，允許驅動裝置藉由單獨箝夾該 mm厚之蓋板而喃合光碟組件。亦且如上文所討論，該光The regionalized CLV is a single data layer disc that is physically imprinted and holographically encoded as described above, and both can often utilize a different tracking set than the CD_R standard wobble track. In particular, the optical disc of the present invention can be constructed in accordance with the present invention by the principle of "ZCLV" format "Zoned Constant Unear Vel〇city" located outside the optical disc area. As explained briefly herein, the (ZCLV) format is well known in the art and is detailed in various industry standards, including the dvd_ram specification (see, for example, ECMA 272, incorporated herein by reference). As shown in the side view 21 above, the ZCLV disc 2 has a data area 2〇2, which is divided into a plurality of areas 204a-204e. Although only five areas are displayed, the actual ZCLV format disc can have a different number of areas. For example, the dvd_RAM ZCLV style has 24 areas in the writable person (four) area. Each area 2〇4a_204e is divided into a plurality of sectors 2〇6. The inner area has fewer sectors than the outer area because the inner area radius is smaller than the outer area radius. The layout of the disc is configured such that the data block slot header information on each of the magnetic stops (i.e., the spiral turns) is radially aligned within each sector. This situation is allowed to be used for each 988l5.doc -45- 1249158 ::==, _ area immediately after the data is used, the disc reader can be in each area of the disc and still dimension (four) domain (four) constant data, on ^ = words In the dimension _ the upper _ 圏 2" s domain... the CD can be rotated at a lower rate. Figure 22 shows a portion of one of the sectors of the ZCLV disc 2 。. As can be seen, most of the tracks 220 Radial configuration in the sector, == in the prefabricated track, the area 222 embossed embossed data block of the field. Each of the magnetic data can be recorded in the wobble track and the wobbled area between the wobbles in the wobble track and the track area 224 as being used as the verification optical disc according to the present invention, and the non-operational structure can be configured. In the wobble pattern and track area 224 of the sector on the optical disc, as shown in Fig. U, because in each sector, the wobble pattern and the track area "A form a continuous performance area" use the ZCLV format test The fixed disc can perform different inspections using the sectors. The embossed imprint head information in the pre-formed track area 222 can be used to store information identifying the verification in the wobbled surface and the track area. The oscillating surface and groove in the oscillating surface and the track area and the embossed data in the pre-fabricated area satisfy the functional requirements of the optical disc reader. As suggested above, the features of various different digitally diverse optical disc (DVD) standards can be advantageously utilized in the implementation of the single data layer embodiment of the present invention. Therefore, the present invention is not limited to existing CD_type discs or cd standards. In addition to the ZCLV format specifically utilized in the majority of the assay light 98815.doc -46-1249158 discussed above, it has become apparent that the smaller structure size and lower wavelength laser developed for the DVD standard allow for higher density non-operational structures. A stereoscopic clock that is more likely to use the CD standard. Adjusting the dvd_ram standard is another important advantage of the invention for the image-inverted disc. Its grain-grain tracking mode (refer to A-27) will be used for the non-operational structure cross-measurement and features on the raft. The tangent resolution is doubled. Even the two-layer data layer DVD format that provides unique features will be discussed further in the letter section. Disc Cover The CD_R reader thus used in Examples 2 and 3 has been modified here by the addition of the focus lens 17 to adjust the focus of the drive when the laser lacking the first surface of the disc of the particular analyte is approaching the refractive substrate. Another method that is often preferred is to adjust the light itself rather than the optical disc reader. In the latter manner, the 'laser refractive member is attached to the proximity side of the disc laser as a cover which acts to refract and thus focus the incident light on the disc-refractive surface. When properly designed, the cover excludes another alternation in the focusing optics of the drive. The use of such a cover plate results in the material presented in Figure 4 without the aid of another focusing lens 17. This way is depicted in Figure 19. Here, by conventional means, the laser light is incident from the bottom of the side cross-sectional view. The optical disc 130 includes a disc substrate 132 and a reflective layer 13 4 '. The non-operational structure 136 is disposed on the reflective layer. The wobble track 13 8 is pressed in the substrate 132 and plated by the reflective layer 134 as indicated. Also shown in the figure is a non-integrated cover plate 140. One embodiment of such a cover is further shown further in the above perspective view in Figure 35. Preferably, the optical disc assembly (disc plus the attachment cover) is sized to approximate the size of the unit 98815.doc -47-1249158, that is, the depth is 12 mm and the diameter is 80 mm. Or 120 mm. However, it is also contemplated that the disc assembly can be varied in size. In these latter cases, regardless of the physical size specification of the disc standard, the assembly must still demonstrate compliance with the necessary optical and mechanical requirements of the drive unit. • Between these requirements, the laser must be properly focused on the disc operating plane. The disc assembly must properly hold the spindle, and the disc assembly must not change much with the standard weight so that the drive motor cannot maintain the proper rotational speed. Example 6 presented here below illustrates that the cover plate made of polycarbonate is approximately M7 mm thick, but otherwise the size is equivalent to a standard i2 〇 disc. The cover sheet is made of polycarbonate as a well-known optical quality utilizing polycarbonate and adapted to the ready-to-use properties of the casting by means of a device. However, as discussed further below, other plastics can be advantageously used to construct a disc cover. A single data layer disc manufactured as in Example 5 has a thickness of 12+/_〇〇5; a single data layer disc manufactured as in Example 6 is approximately 丨^ thick. The external thickness of the physical thickness provided by the Red Book standard (all combinations are ^ _ 15 mm) is the depth of the two cover plates approx. Although this situation does not present an optical problem - maintaining the optical path within the specification - it appears that it may be possible to increase the thickness for this to see the difficulty of clamping the optical disc assembly in the drive 4 center. Even the combination of the disc and the cover exceeds the standard, the weight of the disc. However, 'experientially, we did not expect to find that the clamp and the rotating assembly have no two problems. Now the information in Figure 4 is the use of the optical disc manufactured as in Example 5 and the manufacture of the optical drive according to Example 6. Cover plate set 98815.doc 1249158. The cover plate provides sufficient assistance to focus to exclude the addition of another focus lens to the disc player pickup to produce the information set forth in Figure 40. In a series of separation experiments (not shown), we removed the jaw ring from the disc after attaching the cover, allowing the drive to mate the disc assembly by clamping the mm-thick cover separately. And as discussed above, the light

碟本身可以藉由在DVD_R鑄模中沖壓而減低厚度。 雖然非整合構件目前較佳化為雷射折射光碟蓋板之構 件，可安裝絞鍊或是另夕卜可移冑附接 < 整合構件或是不能 依據所需之使用，非整 移動之整合構件亦可以加以使用 合構件可以反向地（可移除）或是非反向地附接至該光碟The disc itself can be reduced in thickness by stamping in the DVD_R mold. Although the non-integrated component is currently better suited as a member of the laser refracting disc cover, the hinge can be mounted or otherwise attached. The integrated component can not be integrated according to the required use. The member can also be attached to the disc in reverse (removable) or non-reversely.

众生呈現於圖4〇之資料，如例子6製造之非整合聚碳 酉=脂盍板可以反向地固定於如例子5製造之光碟。可以部 分使聚碳酸脂溶解以及使其成為發黏的二至三小滴甲基甲 銅(MEK),被應用於光碟籍夾環142。商業上可使用之塗 漆等級被使用。之後蓋板被溫和地壓印在光碟上大〇 秒。贿永久地將蓋㈣定光碟箝夾環；在組件外徑上， 光碟和蓋板保持緊密地並列但不附接。 &系’先光予和機械需求&需要光碟孝口蓋板以緊密徑向登記 、且口以防止光碟離開圓周；在轉動期間之不正常行為可以 防止伺服機構鎖定尋軌信號。 為確定適當的登記，本發明光碟和匹配之非整合蓋板將 :佳地具有相互傷合及/或相互鎖定之非操作結構。在一 :佳之方式中’該蓋板將和光碟為周圍地重疊。如例子6 造之離形蓋板，缺乏工程化之結構，如下^位於光碟中 988l5.doc -49- Ϊ249158 心 ο 在啟始工作中，CD支架(，，珠寶盒")之塑膠托盤被用以使 早:身料層前向影像、正浮雕倒置光碟不能移動。MEK以 “應用於光碟箝夹裱且蓋板之後置放於頂端以及壓製到 正確位置。支架中的可變形轉轴保持光碟和蓋板為夠緊密 登記以允許成功的組合。這些工作之後續，簡單的、專屬 裝置被改革以完成該登記。 分別如例子5和例子6製造之光碟#蓋板為各具有疊環 144’將箝夾環142箝夾環142由資料區分離。該資料區， 由光碟（和蓋板）-側突出但非另一側，當堆疊時，設計為 使鄰近光碟互相保持足夠距離以防止刮傷。因此，以疊環 組合之光碟和蓋板為相互面對。 MEK在此使用於實驗樣本中，目&方便和快速允許聚碳 酸脂結構附接。其他黏合劑亦可以使用而且需要除了聚碳 酸脂以外的塑膠使用於蓋板及/或光碟。甚至於，MEK使 聚碳酸脂表面結霜；雖然其應用於箝夾環142沒有呈現光 干問題，如其位於資料區外部一樣，假使結合在資料區内 為所需時，較近於光碟外徑，MEK不能容易地使用。為了 那些目的，光學上更合適的黏合劑將為較佳；在此種黏合 劑之間，那些典型用以附接DVD光碟中的分離薄層之黏合 剑（參考下文）可以證實為較佳。一種該黏合劑在美國專利 案唬為第5,879,774號中說明，該文獻在此以提及的方式併 入本文中。尚在審理中以及共同擁有之美國專利申請案號 為第 09/263,972號由 Virtanen所申請取名為”M〇n〇m〇lecuiar 98815.doc -50- 1249158The data shown in Fig. 4 is shown in Fig. 4, and the non-integrated polycarbonate 盍 = grease raft plate manufactured as in Example 6 can be reversely fixed to the optical disc manufactured as in Example 5. The two to three small drops of methyl copper (MEK), which can partially dissolve the polycarbonate and make it sticky, are applied to the optical disk holder ring 142. Commercially available paint grades are used. The cover is then gently stamped on the disc for a maximum of two seconds. The bribe will permanently cover (4) the disc clamp ring; on the outer diameter of the assembly, the disc and cover remain tightly juxtaposed but not attached. The &'s first light and mechanical requirements & requires a compact visor cover for tight radial registration and a port to prevent the disc from leaving the circumference; abnormal behavior during rotation prevents the servo from locking the tracking signal. In order to determine proper registration, the disc of the present invention and the mating non-integrated cover will preferably have non-operating structures that are mutually indented and/or interlocked. In a preferred mode, the cover will overlap the disc. As shown in Example 6, the off-shaped cover plate lacks the engineered structure. The following is located on the CD-ROM. 988l5.doc -49- Ϊ249158 Heart ο In the beginning of work, the CD tray (,, jewel box ") plastic tray was It is used to make the early: body layer forward image, positive relief inverted disc can not move. The MEK is “applied to the disc clamp and placed behind the cover and pressed into the correct position. The deformable shaft in the bracket keeps the disc and cover tightly registered to allow for a successful combination. A simple, proprietary device was reformed to complete the registration. The optical disc # cover plates manufactured as in Examples 5 and 6, respectively, have stacked loops 144' to separate the clamp ring 142 clamp ring 142 from the data zone. The disc (and the cover)-side protrudes but not the other side, when stacked, is designed to keep adjacent discs at a sufficient distance from each other to prevent scratching. Therefore, the disc and the cover plate combined with the stacked loops face each other. MEK is used here in experimental samples to facilitate and quickly allow the attachment of polycarbonate structures. Other adhesives can also be used and require plastics other than polycarbonate for use on covers and/or optical discs. MEK frosts the surface of the polycarbonate; although it is applied to the clamp ring 142 without the problem of light drying, as it is located outside the data area, if it is needed in the data area, it is closer to the disc. Outer diameter, MEK cannot be easily used. For those purposes, an optically more suitable adhesive would be preferred; between such adhesives, those typically used to attach a separate layer of a DVD disc ( Reference may be made to the following. It is believed to be preferred. One such adhesive is described in U.S. Patent No. 5,879,774, the disclosure of which is incorporated herein in The case number is 09/263, 972, which was applied by Virtanen to name it "M〇n〇m〇lecuiar 98815.doc -50-1249158

Adhesion Methods for Manufacturing Microfabricated Multilaminate Devices，，，該文獻在此以提及的方式併入本 文中’呈現另一替代方案於標準黏合劑，可以證實為特別 有利於固定雷射折射蓋板至具有非操作結構之可尋軌光 碟。 聚碳酸脂被選擇於在此舉例證明之蓋板以利用聚碳酸脂 熟知的光學稟直以及裝置調適於其鑄造之存在使用性。然 而，其他塑膠可以優越地使用於建立光碟蓋板。此種塑膠 包含聚甲基丙烯酸、聚乙烯、聚丙烯、聚丙烯酸、聚甲基 丙烯酸酯、聚氣乙烯、聚四氟乙烯、聚苯乙烯、聚碳酸 月曰、聚乙縮醛、p〇lysulf〇nes聚磺、纖維素醋酸鹽、硝酸 纖維素、醋酸纖維素或是其中之混合物。玻璃亦可以使 用0 又口丄又所表 ’’冲I r、、、口 4舟匈议狂地興本發明單一資料 層光碟之操作結構共焦配置。此情形允許後者同時聚焦在 光，之非操作結構與操作結構上。甚至於，當信號元素直 接洛於操作結構内時-由此舉例證明之光碟中，落於擺動 紋道-信號被最大化。 =而’將瞭解地是非操作結構和操作結構（制是尋軌） 作相等之聚焦平面中-能夠滿足該非操作結構和操 構:U共焦而使光碟閲讀機目的組件同時㈣該二結 二碑Γ作結構可以配至在蓋板雷射遠端側(也就是 -先樂接近）而非另外在光碟表面本身上。此方式呈現 98815.doc -51 . 1249158 一些重大優點。 第一’配置非操作結構在塑膠上引人注目地增加用以固 疋非操作結構之化學物，如特定分析物信號元素至表面。 雖然金-硫化黏合劑證實可廣泛調適-如抗體（例子2)之附 著、細胞（例子3)之附著、核酸（例子7)之附接-塑膠呈現可 使用附接化學物之更廣泛選擇。 第二’雖然金表面可以圖樣化以呈現該附接之離散格 鲁 點，塑膠表面甚至可以更容易地衍生以呈現立體定義圖樣 之化學反應群組；這些反應群組圖樣依次加強在立體可尋 址圖樣中之非操作結構的應用。 這些圖樣中之某些圖樣和其優點在共同擁有以及尚在審 理中的美國專利申請案案號為第〇8/888,935號，1997年7月 7曰申請；以及第〇9/12〇,〇49號，1998年7月21曰申請之申 請案中說明；這些文獻在此處以提及的方式併入本文中。 在該文獻中討論之優點為報告跨越廣泛動態範圍之分析物 籲 濃度之圖樣以配置信號元素的能力。 其他立體定義和立體可尋址圖樣容易建立這些優點本 身。例如，以其他基板上之立體可尋址格式如矽晶片或是 玻璃片配置核酸之利用為熟知。 甚至於，置放較近於光碟内徑之特定分析物元素在光碟 動作開始時被讀取；置放於内徑漸遠之特定分析物元素在 漸大轉動延遲之後被讀取。在非均衡分析中，該圖樣容易 地允許動力驗定被執行，而較早之反應時間點因此由配置 光碟更周圍之驗定格點報告。 988I5.doc -52- 1249158 第二’塑膠表面亦可以容易地衍生以呈現所需程度之親 水性’當表面必須一致以似水樣本弄濕時，呈現在金屬表 面之上的優點。另外，表面可以圖樣化為區域呈現變化程 度之親水性和恐水性。 第四，我們已經觀察到跨越光碟表面之流體流量由擺動 紋道本身影響以及由紋道本身阻止。此情形在圖38中實際 說明，其中使用之微串珠為容易造成沿著紋道對齊。光碟 蓋板之平坦表面沒有呈現此種阻礙至跨越其表面的流體均 勻流量。 第五，microfluidic成分可以容易地工程化為塑膠蓋板。 該microfluidics尤其在共同擁有和尚在審理中之申請案序 號為09/064,636號申請案中說明，該文獻在此處以提及的 方式併入本文中。 配置非操作結構在塑膠蓋板上的另一優點為，當該蓋板 為非整體和可由光碟組件移除時，該可尋軌光碟本身可以 再使用。 該蓋板-無論整體或是非整體，可移除或是永遠固定， 具有或是不具有配置在上面的非操作結構-可以作用為其 他優越功能。應該明顯的是本發明光碟反射表面外露於空 間中’與標準光碟反射層114相反。為此原因，非氧化金 屬為較佳地使用於其製造中，雖然鋁或是由薄層塑膠覆蓋 之非氧化金屬可以使用。正外露時，本發明第一表面之特 疋分析物驗疋光碟之反射層13 4受到磨損、灰塵和類似 物，可能衰減由此獲得之信號。塑膠蓋板有利地保護反射 98815.doc -53- 1249158 表面以及保護資訊以免受到環境衰減。 另外。亥蓋板作用為將傳染病和其他引起疾病的媒介與 隔離使用者之 析0 重大好處為病毒媒介如HIV之免疫分 由操作立場，在非操作結構136已經配置在反射層134上 之後的可移除蓋板140之應用，可以壓縮非操作結構以及 驅動其至擺動紋道138中，更進一步將信號元素接近至光 碟非操作結構，以增加信號。 在有利於雷射折射蓋板之建構的塑膠間，聚苯乙烯證實 特別有利：許多現行臨床驗定為在聚苯乙烯表面上執行。 使用於酵-鏈結之免疫吸收物驗定（eusa)和放射免疫檢定 法（RIA)之標準微滴定量碟狀物，由聚苯乙烯製成。參加 在聚苯乙烯表面上臨床驗定之執行的大量豐富經驗；因此 此種驗疋可以谷易地调適為呈現平臺。另外，聚苯乙婦之 精密鑄造目前被實現以及容易地完成。 甚至於，該蓋板亦可以有利地由玻璃而非塑膠或是其他 實質上透明超冷流體、液晶或是包含矽之結晶材料組成。 理所當然瞭解地是，該蓋板140厚度必須調整為考量所 選擇塑膠之折射指數的差異以將雷射正確地劇聚焦在光碟 非操作結構上。該調整在光碟技師之技巧内為理所當然。 亦將瞭解地是，經由蓋板偵測非操作結構_也就是說， 執行第二表面而非第一表面-將有時由增益控制電路驅動 之呈現中獲得益處’如CDRW驅動裝置之大量主要性上發 現〇 98815.doc -54 - 1249158 雖然說明為一致組合，可能以表面圖樣化而言，該蓋板 亦可以具有非一致組合，在徑向平面或是光軸方向或是二 者之方向。 亦可能藉由以薄透明層塑膠將光碟反射表面鍍層而達成 藉由配置非操作結構在塑膠蓋板上所賦予之許多優點，該 非操作結構之後應用在蓋板上（亦即在此多重薄層結構的 最雷射接近表面上）。聚苯乙烯樹脂可以容易地用於此目 鲁 的以及之後原位熱化處理。該樹脂藉由真空配置或是轉動 鍍層加以應用，之後以uv光熱化處理；該處理以聚甲基 丙烯酸在相關技藝中實現，廣泛熟知為，，2p"處理。 土 此後者的方式賦予配置非操作結構，如特定分析物信號 凡素在塑膠上的先前提到之優.點。甚i於，該#式刪除另 外存在於蓋板和光碟金表面之間的界限狀況，允許由更協 調的光之光碟操作平面回轉。然而，因為該層設計為夠薄 以置放實質上與光碟操作結構共焦之該信號元素，該層單 •冑不足以完全輔助聚焦。一合適大小之蓋板則可以另外使 用。 甚至於，該鍍層可以有利地以薄層玻璃而非塑膠或是其 他實質上透明超冷流體、液晶或是結晶材料如石夕組成。 如上文所表示，增加雷射折射蓋板至該光碟以建立重新 儲存第二表面特徵至光碟之光碟組件時，允許額外透鏡口 由光學拾音器移除。呈現在圖40中之資料為使用如例子5 製造之具有如例子6製造之蓋板的光碟而獲得，由仏^ 6200S CD-RW光碟機讀取而不用在例子2和例子3中輔助聚 988l5.doc -55- 1249158 焦之額外聚焦透鏡。 如圖3一1和圖32所顯示，在例子5製造之光碟為用以產生 67 '員不之貝料’具有紋道深度近似100 nm，接近第一表 面债测之最佳化。甚至具有蓋板時，該丨⑽·紋道深度允 許介於信號和背景之間的現成鑑別，㈣辦所顯示之電 押、二所€明’實際說明建構具有可讀取之非操作結構的 單貝料層可尋執光碟之目前方式為非常健全。Adhesion Methods for Manufacturing Microfabricated Multilaminate Devices,, which is hereby incorporated by reference, in its entirety, which is hereby incorporated hereinby Structured trackable disc. The polycarbonate is selected from the cover sheets exemplified herein to take advantage of the optical straightening well known in polycarbonate and the suitability of the device for its casting. However, other plastics can be used to create a disc cover. The plastic comprises polymethacrylic acid, polyethylene, polypropylene, polyacrylic acid, polymethacrylate, polyethylene, polytetrafluoroethylene, polystyrene, polycarbonate, polyacetal, p〇lysulf 〇nes polysulfonate, cellulose acetate, nitrocellulose, cellulose acetate or a mixture thereof. Glass can also be used with 0 and 丄 丄 ’ ’ 冲 I r 、 、 舟 舟 舟 舟 舟 舟 匈 匈 匈 匈 匈 匈 匈 匈 匈 匈 匈 匈 匈 匈 匈 匈 匈 匈 匈 匈 匈 匈 匈 匈 匈This situation allows the latter to focus on the light, the non-operational structure and the operational structure. Even when the signal elements are directly connected to the operating structure - in this exemplified disc, the falling track - the signal is maximized. = and 'will understand that the non-operational structure and the operational structure (the system is the tracking) in the same focal plane - can meet the non-operational structure and operation: U confocal and make the disc reader target components simultaneously (four) the two knot two The monument structure can be placed on the distal end side of the cover laser (ie, the proximity to the music) rather than on the surface of the disc itself. This approach presents 98815.doc -51 . 1249158 with some significant advantages. The first 'configuration non-operating structure significantly increases the plastic on the plastic to immobilize non-operating structures, such as specific analyte signal elements to the surface. Although gold-vulcanized binders have been shown to be widely adaptable - such as attachment of antibodies (Example 2), attachment of cells (Example 3), attachment of nucleic acids (Example 7) - plastic presentation may be more widely available using attachment chemistries. Secondly, although the gold surface can be patterned to present the discrete Glu points of the attachment, the plastic surface can even be more easily derived to present a chemical reaction group of stereo definition patterns; these reaction group patterns are sequentially enhanced in stereoscopic search. The application of non-operational structures in the address pattern. Some of these drawings and their advantages are co-owned and are still pending in the U.S. Patent Application Serial No. 8/888,935, filed July 7, 1997; and No. 9/12, 〇 No. 49, filed July 21, 1998, filed hereby incorporated herein by reference. An advantage discussed in this document is the ability to report patterns of analytes across a wide dynamic range to configure signal elements. Other stereo definitions and stereo addressable patterns make it easy to establish these advantages themselves. For example, the use of nucleic acids in stereoscopically addressable formats on other substrates, such as germanium wafers or glass sheets, is well known. Even a particular analyte element placed closer to the inner diameter of the disc is read at the beginning of the disc motion; the particular analyte element placed at an inner diameter is read after the progressive rotation delay. In the non-equilibrium analysis, the pattern readily allows the power check to be performed, and the earlier reaction time point is therefore reported by the test spot around the configuration disc. 988I5.doc -52- 1249158 The second 'plastic surface can also be readily derivatized to exhibit the desired degree of hydrophilicity' when the surface must be uniform to wet the water sample, presenting an advantage over the metal surface. In addition, the surface can be patterned to show the degree of change in hydrophilicity and water inevitability. Fourth, we have observed that the flow of fluid across the surface of the disc is affected by the wobbled groove itself and by the groove itself. This situation is illustrated in Figure 38, where the microbeads are used to easily align along the track. The flat surface of the disc cover does not exhibit such a barrier to a uniform flow of fluid across its surface. Fifth, the microfluidic component can be easily engineered into a plastic cover. The microfluidics are described in particular in the application Serial No. 09/064,636, the disclosure of which is incorporated herein by reference. Another advantage of configuring the non-operating structure on the plastic cover is that the trackable disc itself can be reused when the cover is non-integral and can be removed by the optical disc assembly. The cover, whether integral or non-integral, removable or permanently fixed, with or without a non-operating structure disposed thereon, can function as a superior function. It should be apparent that the reflective surface of the optical disc of the present invention is exposed to the space 'as opposed to the standard optical disc reflective layer 114. For this reason, non-oxidized metals are preferably used in their manufacture, although aluminum or non-oxidized metals covered by a thin layer of plastic can be used. Upon exposure, the reflective layer 13 of the first surface of the first surface of the present invention is subjected to abrasion, dust and the like, possibly attenuating the signal thus obtained. The plastic cover advantageously protects the surface and protects the information from environmental degradation. Also. The role of the cover plate is to isolate infectious diseases and other disease-causing media from isolated users. A significant benefit is that the immunization of viral media such as HIV is divided into operational positions, after the non-operating structure 136 has been placed on the reflective layer 134. The application of the cover plate 140 removes the non-operative structure and drives it into the wobble track 138, further bringing the signal elements closer to the disc non-operational structure to increase the signal. Polystyrene has proven to be particularly advantageous between plastics that facilitate the construction of a laser refractive cover: many current clinical tests are performed on polystyrene surfaces. A standard microtiter plate for immunosorbent assay (eusa) and radioimmunoassay (RIA) for use in leaven-chains, made of polystyrene. Participate in a wealth of experience in the implementation of clinical validation on the surface of polystyrene; therefore, such testing can be adapted to the presentation platform. In addition, precision casting of polystyrene is currently being implemented and easily completed. Even the cover may advantageously consist of glass rather than plastic or other substantially transparent ultra-cold fluid, liquid crystal or crystalline material comprising tantalum. It is of course understood that the thickness of the cover plate 140 must be adjusted to take into account the difference in refractive index of the selected plastic to focus the laser correctly on the non-operating structure of the disc. This adjustment is taken for granted in the skills of the disc technician. It will also be appreciated that detecting non-operational structures via the cover plate - that is, performing the second surface rather than the first surface - will sometimes benefit from the presentation driven by the gain control circuit - as the bulk of the CDRW drive Sexually found 〇98815.doc -54 - 1249158 Although the description is a consistent combination, the cover may also have a non-uniform combination in the radial plane or the optical axis direction or both directions in terms of surface patterning. . It is also possible to achieve many advantages imparted to the plastic cover by arranging the non-operating structure by plating the reflective surface of the optical disc with a thin transparent layer of plastic, which is then applied to the cover (i.e., multiple layers in this layer). The most laser of the structure is close to the surface). Polystyrene resin can be easily used for this and subsequent in-situ thermal treatment. The resin is applied by vacuum or rotary coating followed by uv photothermal treatment; this treatment is achieved in the art of polymethacrylic acid, widely known as 2p" treatment. The way the latter is given gives the configuration of non-operating structures, such as the specific analyte signal, which is previously mentioned in plastics. In other words, the #-type deletion additionally exists between the cover and the gold surface of the disc, allowing the plane to be rotated by the more coordinated optical disc. However, because the layer is designed to be thin enough to place the signal element that is substantially confocal to the disc operating structure, the layer is not sufficient to fully assist focus. A suitable size cover can be used separately. Even, the coating may advantageously consist of a thin layer of glass rather than plastic or other substantially transparent ultra-cold fluid, liquid crystal or crystalline material such as Shi Xi. As indicated above, the additional lens aperture is allowed to be removed by the optical pickup when the laser refracting cover is added to the disc to create a disc assembly that re-stors the second surface feature to the disc. The data presented in Fig. 40 was obtained using a disc manufactured as in Example 5 having a cover plate as manufactured in Example 6, and was read by a 6^6200S CD-RW disc drive without being assisted in Examples 2 and 3 by an auxiliary 988l5. .doc -55- 1249158 Extra focus lens for focus. As shown in Figs. 3 to 1 and Fig. 32, the optical disc manufactured in the example 5 is used to produce 67's of beryllium material' having a groove depth of approximately 100 nm, which is close to the optimization of the first surface debt measurement. Even with a cover plate, the 丨(10)·grain depth allows for off-the-shelf identification between the signal and the background, (4) the electric charge displayed by the office, and the two instructions are actually constructed to have a readable non-operational structure. The current way in which the single-shell layer can be used to search for optical discs is very robust.

蓋板140為遠擇性地呈現，而驗^袼點為正準備在光碟 上’樣本被加上’、需要顯影該驗定時，執行另一驗定步 驟此後，為準備光碟於讀取，蓋板14〇置放在反射層m 和信號元素136上。 擺動偵測 ^上文所討論，該擺較道為證實特财利於本發明單 -資料層具體實施例中操作和非操作信號之同時和可鑑別 ㈣的尋執結構，為首先加入至光碟標準以允許cd(和之 後的DVD)媒體之使用者導向記錄。如相關標準所陳述， 口語術語化為橘皮書，該擺動由記錄裝置在寫人資料至光 碟期間所偵測；此後’尋軌藉由偵測如此寫入之資料而完 成。特定光碟裝置，主要為品質控制目的而已經製造，具 有能力偵測擺動尋軌結構而不用寫入至光碟。此種裝置的 例子包含由加州Irvine市CD Ass〇ciates公司和瑞典觀_ 市Audio Devel〇pment AB製造的那些光碟。因此，除了使 用於這些特殊化品質控制裝置以外，該標準和許多標準之 存在製作，一般預期該擺動在寫入之後為冗餘。 98815.doc -56- 1249158 因為該擺動典型只在寫入期間偵測，一處理不需要於本 發明大多數製作（以及造成振幅可能干擾特定分析物信號 偵測之雷射脈衝），在CD製造中特殊設計於品質控制目的 之讀寫器在例子2和例子3中使用；此裝置偵測和追蹤該擺 動而不強制雷射以光碟寫入時所需能量加以脈衝。不需要 用以產生圖40資料之額外聚焦透鏡的該Ricoh 6200S CD· RW光碟機被調適為讀取該擺動而不需寫入至該光碟。 信號處理 在例子3中，如上文簡易說明以及在下文詳細說明，該 類比HF信號被饋送至數位示波器以產生在圖13-18顯示之 即時追蹤。相反地，圖40資料首先被擷取、數位化、儲存 在電腦磁碟上，以及此後只藉由解釋性軟體顯示在電腦監 視器上，更進一步在Worthington等人所撰寫，於1999年8 月23日申請之美國專利申請案案號為第09/378,878號（事務 所案號為 BURST-15)之"Methods and Apparatus for Analyzing Nonoperational Data Acquired from Optical Disc”說明以及在Worthington等人所撰寫，於1999年8月23 曰申請之美國專利申請案案號為第60/150·288號（事務所案 號為 BURST· 18Ρ)之 ’’Methods and Apparatus for Optical Disc Data Acquisition using Physical Synchronization Markers’1說明，該申請案文獻在此處以提及的方式併入本 文中。 多重資料層光碟 本發明第二系列具體實施例利用制訂於最近發展之數位 98815.doc -57- 1249158 多樣化光碟（DV)格式之多ί資料層結構。如τ文所詳細討 論’該DVD格式特別良好地適合於提供光碟幾何亦即尋軌 設計，允許光料IU线與非操料構產生之信號同時操 取以及鑑別。 現在參考圖24,顯示典型雙^VD格式光碟。藉由此處 之傳統’雷射光由下方入射。光碟包括雷射接近基板 282、半反射層284、間隔層286、反射層288和雷射遠端基 板 290。 接近基板282包括透明光學材料如聚碳酸脂，具有所選 擇折射指數，用以輔助將雷射光束聚焦在二層資料之一 側接近基板282可以由類似於上文用於製造CD_R格式光 碟處理之注射鑄造處理加以製造。接近基板282為典型地 以/〇著螺旋軌配置之資料加以浮凸壓印。這些資料為典型 地視為存在於二層光碟之，，層0，，中。 接近基板282之該含有資料表面以半反射層 284加以鍍 層，包括非常薄之鍍層材料如矽、金、鋁、銀、銅或是其 中之合金，用以反射一些光以及傳輸一些光。雖然反射性 可以容納一個範圍，但半反射層284典型地具有近似3〇% 之反射性。因此’半反射層284可以具有大約2〇%-40%之 反射性，較佳地是25%-35%，最佳地是大約3〇%。 遠端基板290包括如聚碳酸脂之材料，可以螺旋資料軌 加以矯造。因為雷射光束將不通過遠端基板29〇，所以其 光學特徵為不重要。遠端基板29〇可以由注射鑄造處理加 以製造，如上文所述。 98815.doc -58- 1249158 遠端基板2 9 0以在螺旋資料軌中的資料加以浮凸應印， ㈣可師層〇之„賴執平行運行（㈣由光碟内部部 刀至外部部分）或是在層0之螺旋資料軌之反方向（亦即由光 碟外部部分至内部部分）。浮凸塵印在遠端基板290中的資 料視為存在於二層光碟之，，層丨，，中。 通端基板290之該含有資料表面以反射層咖加以鑛層， 包括薄層之任何材料如金、銘、銀、銅。反射層挪典型 * 土也具有设計為可能接近層0之反射性。當由-層交換至另 I層讀取時，此方式可以藉由自動增益控制強制再調整而 完成，·增益之此種變化可以反向影響尋執。對此理由而 言，雙層光碟之層1最經常具有遠低於70%之反射性。 間隔286提供40至70微米之空間介於雙層光碟之層丨與層 〇之間，以及作用為結合接近基板282和半反射層284至遠 端基板290和反射層288。間隔286典型地包括具有接近於 製造接近基板2 8 2之材料折射指數的折射指數之光學膠黏 ’劑。 在使用時，DVD閱讀機可以將雷射光束聚焦在半反射層 284—側上，以讀取層〇中之資料，或是在反射層288上: 以讀取層1中之資料。DVD光碟多層特性和DVD閱讀機共 存的雙焦點使得DVD特別良好地適合使用於本發明：由光 碟操作結構所佔據之平面在這些具體實施例中，實體上可 以和特定分析物元素所佔據之平面分離，加強二類型資料 之同時可鑑別擷取。 因此，在一具體實施例中，非操作結構和資料層〇共 98815.doc •59- 1249158 焦，該光碟尋軌和其他操作結構位於資料層1。相反地， 在另一具體實施例中，非操作結構和資料層丨共焦，且該 光碟尋執和其他操作結構位於資料層0。在另一替代方案 中 驗疋元素為配置間隔層286中，實質上與二資料層之 一層共焦。 這些原理中之一些原理藉由參考圖25顯示之具體實施例 加以實際說明。圖25呈現DVD類型雙資料層驗定光碟3〇〇 之分解側視圖。該光碟實質上沿著在典型雙曾Dvd光碟中 由間隔層286定義之平面拆開。 光碟300包括二部分··主部分3〇2和蓋板部分3〇3。該部 分可以永久地固定於另一部分，可以分離和組合或是可以 分離和反向地組合。在這些建構中之任何建構中，在光碟 頃取之前’該蓋板部分303組合在主部分3〇2之外部驗定區 域306之上。開孔3〇8和區域304為如此定大小以允許二件 之緊貼和可靠之安裝。 主邛为302之外部驗定區域306包括單一資料層區域，非 刼作結構為配置在該區域上。類似於上文呈現之單一資料 層具體貫施例，外部驗定區域3〇6以擺動紋道（沒有顯示）加 以浮凸壓印，或是其他合適尋軌結構，使用於提供尋軌資 訊至光碟閱讀機。依據DVD標準，以及和上文呈現之單一 資料層具體實施例相反，該擺動紋道可以為前向影像或是 反向影像紋道。如所提到一般，可以使用格式。 主部分302亦包括内部資料區域3〇4。内部資料區域3〇4 以類似於正常雙曾DVD光碟之方式加以格式化。程式和資 98815.doc -60- 1249158 料可以儲存在光碟此區域之層〇及/或層1。 特別是，内部資料區域304較佳地包含指引光碟閱讀機 調整其焦點以校正該資料層而讀取呈現在驗定區域306中 之特定分析物信號。甚至於，内部資料區域304可以儲存 用以調整韌體或是驅動裝置晶片組”快閃”元件之資料，如 需要地允許驅動裝置正確地讀取和解譯特定分析物信號。 蓋板部分303較佳地包括透明光學材料如聚碳酸脂、聚 • 甲基丙烯酸或是玻璃，選擇為將光碟300操作結構（例如擺 動紋道）偵測以及信號元素偵測加以最佳化。 將為明顯地是，在相關技藝技巧内之良好變化包含將非 操作、、Ό構被至在區域306之層〇或層1，或是在該二層上， 貫體地由該驗定平面分離尋軌結構，該結構可能本身缺乏 尋執結構或是組合。另外，該驗定可以在光碟組合之前藉 由沈澱該信號元素在蓋板部分3〇3上的雷射遠端表面上= 在蓋板部分303上執行。 熟知相關技藝之人士而言，為明顯地是，具有許多微 小變化可以在此具體實施例中製成。例如，假使大量資料 或是程式為解釋驗定結果所需要時，内部資料區域3〇4可 以具有寫在層〇及層1上的資料，而不用改變外部驗定區域 306層0擺動紋道和驗定結果（亦即信號元素）。 多層DVD格式之另一組優點可以藉由參考圖26而瞭解， 該圖為依據本發明建立之另一多層具體實施例驗定光碟之 橫斷面圖。藉由此處之傳統，雷射光由下方入射。 光碟320包括位於間隔層324中的通道322。驗定可以藉 98815.doc -61 - Ϊ249158 由將測試之材料經由開孔325導入至通道322而執行，該開 孔位於光碟雷射遠端側上。當執行驗定時，信號元素為配 置在光碟320層1之反射層326上。 光碟320層1以擺動紋道加以浮凸壓印，提供光碟閱讀機 最小操作需要。光碟320層0包含讀取驗定光碟和解譯該結 果所需要之資料和程式。 對熟知相關技藝之人士而言，為明顯地是，多數驗定可 以藉由使用分離通道322而在單一光碟上執行，各通道設 計為處理不同驗定。另外，將為明顯地是，通道322在間 隔324内的位置可以變化。例如，通道322可以鄰近層〇而 非層1，或是可以粗略地以間隔324定中心。在這些案例其 中之一案例中，信號元素置放於通道322内為執行驗定結 果，可以在聚焦於呈現在光碟320層1中非操作結構上之雷 射回轉路徑中加以偵測。 1998年4月21日申請之共同擁有及上在審理中之申請案 序號為09/064,636號之文獻在此處以提及的方式併入本文 中’說明各種不同通道化及其他三維驗定光碟之變化。 圖27顯示類似於圖26之驗定光碟320的側面橫斷面圖。 雷射將由下方入射。在圖27之光碟330中，通道322為往光 碟外部定位，使光碟3 3 0中央部分成為”標準”二層光碟。 光碟330層0劃分為二區段。層〇區段334儲存資料或是程 式，如上文所說明。層0區段336包括具有光學性質可以和 區段334光學性質不同之透明材料。在一較佳具體實施例 中，層0區段336之光學性質為最佳化於使雷射光束聚焦在 988I5.doc -62- 1249158 光碟非操作結構上，以及最佳化於谓測通道322中的信號 元素。 :考圖28,依據本發明原理建立之另—雙層具體實施例 驗定光碟以類似側面橫斷面圖加以顯示。光碟“Ο可使用 於DVD閱讀機或是CD_R閱讀機。光碟34〇層〇依據dvd格 式配置。在光碟340層〇上編碼之資料可以由標準1)¥]〇播放 機讀取。光碟340層1依據（^次格式編碼，所以可以使用 較廣之磁軌節距以及低密度配置資料。資料可以配置在光 碟州層丨中央部分342。層丨之驗定部分以擺動紋道加以浮 凸壓印以滿足光碟閱讀機操作需求，以及鄰近通道346， 為用於執行驗定，如上文所說明。 圖29顯示二層光碟之另一具體實施例。在光碟35〇上， 二層光碟層1之資料和操作結構由全像352加以提供。全像 352類似於圖20之全像194，所以出現於光碟閱讀機之在全 像352上編碼的操作結構和資料將定於影像平面354上，該 平面相對於全像3 5 2可以為雷射遠端或是雷射接近。通道 356為用於執行驗定，所以信號元素可以配置在光碟35〇之 間隔層358内。這些信號元素在聚焦於全像352操作結構上 之雷射光束的回轉路徑中為可偵測。 對熟知相關技藝之人士為明顯地是類似全像352之全像 可用以提供層1在幾乎上述之任何雙層光碟中。如顯示， 影像平面354相對於全像352表面為雷射接近，所以信號元 素將出現為直接配置在層i表面上，或是在由全像352模擬 的擺動紋道内。 98815.doc -63- 1249158 更進一步對熟知相關技藝之人士為明顯地是參考二層光 不之s午多上述之具體實施例可以容易地延伸為使用具 有多於二層之多層光碟。例如，用於執行驗定之通道可二 夕層光碟之各層之間，而各層（除層〇以外）提供光碟閱 頃機所需要的任何操作結構。 下列例子以解釋方式而非以限制方式提供。 例子1 _ * 口於特定分析物驗定之可尋執前向影像正浮雕倒置光碟 之製造 由Cinram公司（加州安那罕市）製造之包含CD_R格式擺動 紋道影像的未打孔父片，為藉由標準程序基體化為CD_R 母片。簡易地是，該電鑄為藉由Digital 公司（㈣The cover plate 140 is displayed in a remote manner, and the inspection point is ready to be 'sample added' on the optical disc, and the inspection timing needs to be developed, and another verification step is performed thereafter, in order to prepare the optical disc for reading, the cover is The plate 14 is placed on the reflective layer m and the signal element 136. Swing Detection ^ As discussed above, the swaying is to verify that the operation and non-operational signals in the single-data layer embodiment of the present invention are simultaneous and identifiable (four) of the homing structure, for the first time to join the optical disc standard To allow users of cd (and subsequent DVD) media to direct the recording. As stated in the relevant standards, the spoken term is translated into a Orange Book, which is detected by the recording device during the writing of the person's data to the disc; thereafter the tracking is accomplished by detecting the data so written. The specific optical disc device, which has been manufactured primarily for quality control purposes, has the ability to detect the wobble tracking structure without writing to the disc. Examples of such devices include those manufactured by CD Ass〇ciates of Irvine, Calif., and by Audi Develpment AB of Sweden. Therefore, in addition to being used for these specialized quality control devices, the existence of this standard and many standards is generally expected to be redundant after writing. 98815.doc -56- 1249158 Because this wobble is typically detected only during writing, a process that does not require most of the fabrication of the present invention (and laser pulses that cause amplitudes that may interfere with the detection of a particular analyte signal) is manufactured at CD. The readers specifically designed for quality control purposes are used in Examples 2 and 3; the device detects and tracks the wobble without forcing the laser to pulse the energy required to write the disc. The Ricoh 6200S CD·RW disc drive that does not require the additional focus lens used to generate the Figure 40 data is adapted to read the wobble without writing to the disc. Signal Processing In Example 3, the analog HF signal is fed to a digital oscilloscope to produce an instant trace as shown in Figures 13-18, as briefly described above and as detailed below. Conversely, the data in Figure 40 was first captured, digitized, stored on a computer disk, and then displayed on a computer monitor only by interpretive software, further written by Worthington et al., August 1999. The U.S. Patent Application Serial No. 09/378,878, filed on Jun. 23, filed on file No. No. No. No. No. No. No. No. No. No. No. No. No. No. No. No. No. No No No No No No No No No No No No No No No No No No No No No No No No No No No No No No No No No No No No No No No No No No No No No No No No No No No No No No No No No No No No No ''Methods and Apparatus for Optical Disc Data Acquisition using Physical Synchronization Markers'1, US Patent Application No. 60/150.288, filed on August 23, 1999. The application document is hereby incorporated by reference in its entirety. Multi-data layer optical disc The second series of specific embodiments of the invention utilizes the recently developed digital number 98815.doc -57-1249158 Diversified Optical Disc (DV) The format of the data layer structure. As discussed in detail in the article τ, 'The DVD format is particularly well suited for providing optical disc geometry, that is, tracking The light IU line is allowed to simultaneously acquire and identify the signal generated by the non-proper structure. Referring now to Figure 24, a typical dual VD format optical disc is shown. The conventional 'laser light here is incident from below. The optical disc includes a laser. Proximity to substrate 282, semi-reflective layer 284, spacer layer 286, reflective layer 288, and laser distal substrate 290. Proximity substrate 282 includes a transparent optical material, such as polycarbonate, having a selected index of refraction to aid in focusing the laser beam Accessing the substrate 282 on one side of the two-layer data can be fabricated by an injection casting process similar to that used above for fabricating CD_R format disc processing. The proximity substrate 282 is embossed with embossing typically with/or the spiral track configuration. These materials are typically considered to be present on a two-layer disc, layer 0, in. The surface containing the substrate 282 is coated with a semi-reflective layer 284, including very thin coating materials such as tantalum, gold, aluminum. , silver, copper or alloys thereof to reflect some light and to transmit some light. Although the reflectivity can accommodate a range, the semi-reflective layer 284 typically has approximately 3 The reflectivity of 〇%. Thus the 'semi-reflective layer 284 may have a reflectivity of about 2% to 40%, preferably 25% to 35%, and most preferably about 3%. The remote substrate 290 includes as The material of polycarbonate can be modified by spiral data track. Since the laser beam will not pass through the distal substrate 29, its optical characteristics are not important. The distal substrate 29A can be fabricated by injection casting processing as described above. 98815.doc -58- 1249158 The remote substrate 209 is embossed by the data in the spiral data track, and (4) the slab layer is slid in parallel ((4) from the internal part of the disc to the outer part) or It is in the opposite direction of the spiral data track of layer 0 (that is, from the outer portion to the inner portion of the optical disc). The data embossed on the remote substrate 290 is considered to exist in the second layer of the optical disc, layer 丨, , The surface of the substrate 290 containing the data surface is provided with a layer of reflective material, including any material such as gold, inscription, silver, and copper. The reflective layer typically has a design that is designed to be close to the reflection of layer 0. When switching from layer to layer I, this method can be done by forced re-adjustment by automatic gain control. This change in gain can adversely affect the search. For this reason, double layer Layer 1 of the disc most often has a reflectivity well below 70%. Space 286 provides a space of 40 to 70 microns between the layers of the dual layer disc and the layer, and acts to bond the substrate 282 and the semi-reflective layer. 284 to the remote substrate 290 and the reflective layer 288. The spacing 286 is typical An optical adhesive agent having a refractive index close to that of a material that is close to the substrate 28 2 is used. In use, the DVD reader can focus the laser beam on the side of the semi-reflective layer 284 to read the layer. The data in the file, or on the reflective layer 288: to read the data in layer 1. The dual focus of the DVD disc multi-layer feature and the DVD reader makes the DVD particularly well suited for use in the present invention: by the optical disc operating structure The plane of occupancy, in these embodiments, can be physically separated from the plane occupied by a particular analyte element, and the two types of data can be enhanced while discriminating. Thus, in one embodiment, the non-operational structure and data layer 988 a total of 98815.doc • 59-1249158 coke, the optical disk tracking and other operational structures are located in the data layer 1. Conversely, in another embodiment, the non-operational structure and the data layer are confocal, and the optical disk is searched And other operational structures are located at data layer 0. In another alternative, the verification element is disposed in the spacer layer 286, substantially confocal with one of the two data layers. One of these principles The principle is illustrated by reference to the specific embodiment shown in Fig. 25. Fig. 25 is an exploded side view showing the DVD type double data layer verification optical disc 3〇〇. The optical disc is substantially separated by a spacer layer in a typical double-dvd disc. The plane defined by 286 is disassembled. The optical disc 300 comprises two parts, a main part 3〇2 and a cover part 3〇3. This part can be permanently fixed to another part, can be separated and combined or can be combined separately and in reverse. In any of these constructions, the cover portion 303 is combined over the outer verification region 306 of the main portion 3〇2 before the disc is taken. The apertures 3〇8 and 304 are so sized. Allows two pieces to be snug and reliable to install. The external verification area 306 of the main frame 302 includes a single data layer area on which the non-active structure is disposed. Similar to the single data layer embodiment presented above, the external verification area 3〇6 is embossed with a wobble track (not shown) or other suitable tracking structure for providing tracking information to CD reader. In contrast to the DVD standard, and in contrast to the single data layer embodiment presented above, the wobble track can be a forward image or a reverse image track. As mentioned, the format can be used. The main portion 302 also includes an internal data area 3〇4. The internal data area 3〇4 is formatted in a manner similar to a normal dual DVD. The program and resources 98815.doc -60- 1249158 can be stored in the layer and/or layer 1 of this area of the disc. In particular, internal data area 304 preferably includes a directing optical disc reader to adjust its focus to correct the data layer to read a particular analyte signal present in assay area 306. In even, the internal data area 304 can store information to adjust the firmware or "flash" components of the driver chipset, as needed to allow the drive to properly read and interpret specific analyte signals. The cover portion 303 preferably includes a transparent optical material such as polycarbonate, polymethacrylic or glass that is selected to optimize the operation of the optical disc 300 (e.g., wobble track) and signal element detection. It will be apparent that a good variation within the relevant art skill includes the incorporation of a non-operational structure into the layer or layer 1 of the region 306, or on the second layer, from the verification plane. Separating the tracking structure, which may itself lack a look-ahead structure or combination. Alternatively, the verification can be performed on the cover distal end surface 303 by depositing the signal element on the cover distal portion 3〇3 prior to the disc assembly. It will be apparent to those skilled in the art that many variations can be made in this particular embodiment. For example, if a large amount of data or a program is required to interpret the verification result, the internal data area 3.4 may have data written on the layer 层 and layer 1 without changing the external verification area 306 layer 0 wobble track and The result of the verification (ie the signal element). Another set of advantages of the multi-layered DVD format can be understood by reference to Figure 26, which is a cross-sectional view of a verification optical disc in accordance with another multi-layered embodiment of the present invention. With the tradition here, laser light is incident from below. The optical disc 320 includes a channel 322 located in the spacer layer 324. The verification can be performed by introducing 98815.doc -61 - Ϊ249158 from the material to be tested via opening 325 to channel 322, which is located on the distal side of the disc laser. When the timing is performed, the signal element is disposed on the reflective layer 326 of the layer 320 of the optical disc 320. The layer 320 of the optical disc 320 is embossed with a wobble track to provide a minimum operational need for the optical disc reader. The layer 320 of the disc 320 contains the data and programs needed to read the verification disc and interpret the result. It will be apparent to those skilled in the art that most assays can be performed on a single disc by using separate channels 322, each designed to handle different assays. Additionally, it will be apparent that the position of the channel 322 within the space 324 can vary. For example, channel 322 can be adjacent to layer 〇 instead of layer 1, or can be roughly centered at interval 324. In one of these cases, the signal elements are placed in channel 322 to perform the verification results and can be detected in a laser slewing path that is focused on the non-operational structure presented in layer 1 of the disc 320. The co-owned and pending application filed on April 21, 1998, the serial number of which is incorporated herein by reference in its entirety, Variety. Figure 27 shows a side cross-sectional view of the optical disc 320 similar to that of Figure 26. The laser will be incident from below. In the optical disc 330 of Fig. 27, the channel 322 is positioned outside the optical disc so that the central portion of the optical disc 300 becomes a "standard" two-layer optical disc. The layer 330 of the disc 330 is divided into two sections. The layer section 334 stores data or procedures as explained above. Layer 0 section 336 includes a transparent material having optical properties that may differ from the optical properties of section 334. In a preferred embodiment, the optical properties of the layer 0 section 336 are optimized to focus the laser beam on the 988I5.doc -62-1249158 optical disc non-operational structure, and optimized to the pre-measurement channel 322. Signal element in . Reference to Figure 28, another embodiment of a two-layer embodiment constructed in accordance with the principles of the present invention. The optical disk is shown in a similar side cross-sectional view. The disc "can be used for DVD readers or CD_R readers. The disc 34 layer is configured according to the dvd format. The data encoded on the 340 layer of the disc can be read by the standard 1) ¥] player. 340 layer of the disc 1 According to (^ sub-format coding, it is possible to use a wider track pitch and low-density configuration data. The data can be configured in the central part 342 of the disc state layer. The verification part of the layer is embossed by the wobble track. Printing to meet the operational requirements of the optical disc reader, and adjacent channel 346, for performing the verification, as explained above. Figure 29 shows another embodiment of a two-layer optical disc. On the optical disc 35, the second optical disc layer 1 The data and operational structure are provided by a full image 352. The full image 352 is similar to the full image 194 of Figure 20, so that the operational structure and data encoded on the full image 352 of the optical disc reader will be placed on the image plane 354. The plane may be remote to the laser or close to the laser with respect to the hologram 325. The channel 356 is for performing the verification, so the signal elements may be placed in the spacer layer 358 of the disc 35. These signal elements are in focus. Yu Quan The slewing path of the laser beam, such as the 352 operational structure, is detectable. A person skilled in the art will be apparently a holographic image similar to hologram 352 to provide layer 1 in virtually any of the dual layer discs described above. As shown, the image plane 354 is laser close to the surface of the hologram 352, so the signal elements will appear to be placed directly on the surface of the layer i, or within the wobble track simulated by the hologram 352. 98815.doc -63- 1249158 Further to those skilled in the art, it is apparent that reference is made to a second layer of light. The above-described embodiments can be easily extended to use a multilayer optical disc having more than two layers. For example, a channel for performing verification. The layers of the disc can be placed between the layers, and the layers (except the layer) provide any operational structure required for the optical disc reader. The following examples are provided by way of explanation and not limitation. Example 1 _ * Manufactured by the Cinram Corporation (Annaham, Calif.), an unpunched parent film containing the CD_R format wobbled image. To be a CD_R master by standard procedures. Simply, the electroforming is done by Digital ((4)

Hempstead市）製造之電鑄系統中之鎳氣基磺酸鹽槽令執 行。 將忒母片加以β潔、擦亮p〇Hshed和打孔，之後直接使 •肖㈣模以製造具有㈣結構為正浮雕之前向影像螺旋紋 道的倒置光碟。由細⑻機器公司（瑞典市）製造之 NETSTAL鑄造機器和瑞典AWM公司創建之cd_r鑄模為用 以在ΕΧΙΜΡ0 s.R.0(布拉格，捷克共和國）產生光碟。注射 禱造機器之鑄造參數被調整以加強鑄模中的高度通風，而 正確地再生該紋道。用以鑄模光碟之聚碳酸脂由Bayer Plastics公司生產。 該聚碳酸脂光碟基板之後以金使用First Light Technologies緬因州以⑺市）公司製造之喷鍍金屬化合器加 988I5.doc -64- 1249158 以金屬化。 如圖10-12之AFM量測所顯示，這些光碟深度近似170 nm而具有近似1 ·6 μηι之磁軌節距。 例子2 在可尋執光碟上之IgG-特定免疫分析格點之建構 單一資料層、第一表面、前向影像/正浮雕擺動光碟為 依據例子1加以製造。光碟之金表面之後如下衍生以建構 φ 特定於光碟之驗定格點能夠偵測血液樣本中之人體IgG。 2 mg N-[6-(維生素Η氨基）己基]-3’-(2’-比啶基過硫基）丙 醯胺（’，維生素 H-HPDP’，）(Pierce，Rockford，IL;貨品號為 97032461號）之可分量為溶解於2 ml之二甲基甲醯胺。在4 個意圖之驗定格點中的各格點上，各以距離光碟中心相同 之半徑加以定位，10 μΐ之維生素H-HPDP溶液用移液管移 動。該光碟在室溫下被培養2小時，以及之後以50 mM磷 酸鹽緩衝器（pH 7)清洗。 鲁 接著，10 μΐ之 streptavidin溶液（Monobind、Costa Mesa， CA ;貨品96-001/MF ; 2 mg/ml)被用移液管移動在相同驗 定點上。該光碟在室溫下被培養1小時，以及之後以50 mM鱗酸鹽緩衝器清洗。The nickel gas sulfonate tank in the electroforming system manufactured by Hempstead) was implemented. The mother piece is subjected to β cleaning, polishing p〇Hshed and punching, and then the ?(4) mode is directly used to manufacture an inverted optical disk having a (four) structure for positive embossing to the image spiral track. The NETSTAL casting machine manufactured by Fine (8) Machine Company (Sweden City) and the cd_r mold created by Swedish AWM Company were used to produce CDs in ΕΧΙΜΡ0 s.R.0 (Prague, Czech Republic). The casting parameters of the injection prayer machine are adjusted to enhance the high degree of ventilation in the mold while properly regenerating the track. The polycarbonate used to mold the optical disc was produced by Bayer Plastics. The polycarbonate disc substrate was then metallized with gold using a spray metallizer manufactured by First Light Technologies, Maine, Inc. (7) City, 988I5.doc -64-1249158. As shown by the AFM measurements in Figure 10-12, these discs have a depth of approximately 170 nm and a track pitch of approximately 1.7 μm. Example 2 Construction of an IgG-specific immunoassay grid on a searchable disc A single data layer, a first surface, a forward image/positive embossed oscillating disc was manufactured according to Example 1. The gold surface of the disc is then derived as follows to construct a φ disc-specific spot to detect human IgG in the blood sample. 2 mg N-[6-(vitaminium amino)hexyl]-3'-(2'-pyridylperthio)propanamide (', vitamin H-HPDP',) (Pierce, Rockford, IL; goods No. 97032461) can be dissolved in 2 ml of dimethylformamide. At each of the four intent checkpoints, each was positioned at the same radius from the center of the disc, and 10 μL of the vitamin H-HPDP solution was pipetted. The disc was incubated at room temperature for 2 hours and then washed with a 50 mM phosphate buffer (pH 7). Lu Next, 10 μΐ of streptavidin solution (Monobind, Costa Mesa, CA; product 96-001/MF; 2 mg/ml) was pipetted at the same test point. The disc was incubated for 1 hour at room temperature and then washed with a 50 mM sulphate buffer.

Biotinylated goat 抗-人體 IgG 由 Chemicon International 公 司（加州Temecula市；親和力淨化，貨品47797017)獲得。5 μΐ之可分量被用移液管移動在4個驗定格點中的各格點 上。該光碟在室溫下被培養1小時，之後以50 mM磷酸鹽 緩衝器（pH 7)清洗。 98815.doc -65- 1249158 完成之驗定格點的幾何在圖7A中示意。維生素H-HPDP 70形成光碟表面之上的分子層，以金-硫化與格（座標）黏合 劑接合至光碟金表面（Au)。Streptavidin 72，其各分子可 以在高親和力下接合4維生素Η分子，形成次層。 Biotinylated goat抗-人體IgG 76，在驗定格點上賦予驗定 特定性，之後尤其維生素Η —部份74接合至固定 streptavidin 72。該goat抗-人體IgG在允許其固定之位置上 biotinylated，而不干擾抗原（人體IgG)接合。 如此衍生之光碟之後用以驗定人體血液IgG之呈現。 100 μΐ人體血液之樣本由正常志願者抽出。1〇〇 μΐ樣本 之可分量使用磷酸鹽-緩衝器之鹼鹽（，，PBS，，）稀釋10倍。二 種另外10 : 1系列稀釋相同地執行。各一樣本之10微升（μΐ) 可分量-也就是說，未稀釋血液之可分量和3系列稀釋血液 樣本之可分量-為分開和單獨置放在4光碟驗定格點其中之 一驗定格點上。 該光碟在密閉潮濕器室内之氮氣下以室溫培養2小時。 該光碟之後以PBS清洗。 為了發展IgG-特定之驗定，也就是說，使其適合於報告 在應用之樣本中IgG之呈現，5 μ1(160 pg)之MagaBeadsTM goat抗人體 lgG(Fc)(加州 San Leandro市 Cortex Biochem公 司；貨品7A2201 A)被作成記號在4驗定格點中之各格點 上，以及該光碟以50 mM磷酸鹽緩衝器（pH 7)清洗以及之 後以蒸顧水清洗。 在由血液捕獲IgG以及以抗-人體IgG MagaBeadsTM發展 98815.doc -66- 1249158 之後的驗定格點之幾何為在圖冗中加以示意。已經呈現在 血液樣本（为析物）中之IgG 78*bi〇tinylated抗-人體“^ % 加以接合，固定在該驗定格點上。該人體IgG 78之後更進 一步作用為固定抗_人體IgG MagabeadsTM 79。Biotinylated goat anti-human IgG was obtained from Chemicon International (Temecula, Calif.; Affinity Purification, 47797017). The 5 μΐ component is moved by pipettes at each of the four test grid points. The disc was incubated at room temperature for 1 hour and then washed with a 50 mM phosphate buffer (pH 7). 98815.doc -65- 1249158 The geometry of the completed checkpoint is illustrated in Figure 7A. Vitamin H-HPDP 70 forms a molecular layer above the surface of the disc and is bonded to the gold surface of the disc (Au) with a gold-vulcanized and lattice (coordinate) adhesive. Streptavidin 72, each molecule can bind 4 vitamins and molecules to form a sublayer with high affinity. Biotinylated goat anti-human IgG 76 confers specificity on the assay grid, after which in particular vitamin Η - part 74 is conjugated to fixed streptavidin 72. The goat anti-human IgG is biotinylated at a position that allows it to be immobilized without interfering with antigen (human IgG) binding. The disc thus derived is used to verify the presentation of human blood IgG. A sample of 100 μΐ human blood was drawn from normal volunteers. The fraction of the 1 〇〇 μΐ sample was diluted 10 times using the phosphate-buffer base salt (, PBS, ,). Two additional 10: 1 series dilutions were performed identically. Each of the 10 microliters (μΐ) of the same component - that is, the fraction of undiluted blood and the component of the 3 series of diluted blood samples - is separately and separately placed in one of the 4 disc verification grids. Point. The disc was incubated at room temperature for 2 hours under nitrogen in a closed humid chamber. The disc is then washed with PBS. To develop an IgG-specific assay, that is, to make it suitable for reporting the presence of IgG in a sample of the application, 5 μl (160 pg) of MagaBeadsTM goat anti-human lgG (Fc) (Cortex Biochem, San Leandro, Calif.) The product 7A2201 A) was marked as a mark at each of the 4 test grid points, and the disc was washed with a 50 mM phosphate buffer (pH 7) and then rinsed with steam. The geometry of the assay grid after the capture of IgG by blood and the development of anti-human IgG MagaBeadsTM 98815.doc -66-1249158 is illustrated in the redundancy. The IgG 78*bi〇tinylated anti-human "^%", which has been present in the blood sample (as an extract), is ligated and immobilized on the assay grid. The human IgG 78 further acts as a fixed anti-human IgG MagabeadsTM 79.

MagabeadsTM為商業上可使用之球體乳膠可磁化粒子_和不 同種類接合之一部分，如此處之以“抗-人體IgG或是另外 和允許定製共軛之反應群組加以預先共輛。 _ 5亥光碟被乾燥化，以及其表面之後由光線和原子力微觀 (AFM)加以視覺化。圖8為加入人體血液和抗體·共軛球體 之後，由一部分IgG特定第一表面特定分析物可尋執驗定 光碟之光微觀檢視捕捉之視頻影像。圖9和圖1〇為免疫特 定附著於該光碟之單一膠乳球體的AFM影像，放大率為稍 高於圖8所使用之放大率，而總結在影像擷取期間由afm 觀察之測定大小。圖11為免疫特定附著於第一表面可尋執 人體IgG-特定光碟以及呈現在相同領域之二膠乳球體的 • AFM影像，而總結在影像擷取期間由AFM觀察之測定大 /Jn 〇 例子3 在RBC-特定可尋執免疫分析光碟上之人體紅血球的電子 偵測和特徵化 單一資料層、第一表面、前向影像/正浮雕擺動光碟為 依據例子1加以製造。光碟之金表面之後如下衍生。 2 mg N-[6_(維生素Η氨基）己基]_3，_(2，_比啶基過硫基）丙 醯胺（，，維生素 H-HPDP，，）(Pierce，Rockford，il;貨品號為 98815.doc -67- 1249158 97032461號）之可分量為溶解於2 ml之二甲基甲醯胺。在4 個意圖之驗定格點中的各格點上，各以距離光碟中心相同 之半徑加以定位，10 μΐ之維生素H-HPDP溶液為用移液管 移動。該光碟在室溫下被培養2小時，以及之後以50 mM 磷酸鹽緩衝器（pH 7)清洗。 接著，10 μΐ 之 streptavidin 溶液（Monobind、Costa Mesa， CA ;貨品96-001/MF ; 2 mg/ml)被用移液管移動在相同驗 定點上。該光碟在室溫下被培養1小時，以及之後以50 mM磷酸鹽緩衝器清洗。 單系老鼠抗-人體glycophorin A抗體（加州Carpinteria市 Dako公司；貨品113)為如下加以biotinylated。抗體之100 μΐ可分量與0.1 mg之α-維生素Η，ω-Ν-脛基丁二醢胺酯之 聚（乙烯乙二醇碳酸酯（，，Bio-PEG-NHS，，）（AL 州 Huntsville 市Shearwater公司；貨品PT-028-27)為100 μΐ之填酸鹽緩衝 器（pH 7)以及允許反應1小時。維生素Η-共軛抗-人體 glycophorin A為相對於相同緩衝器（透析MWCO = 30,000) 加以透析整夜。 該透析之之維生素H-共輛抗-人體glycophorin A抗體用 移液管移動在光碟上的streptavidin-鍍層驗定點上以及該 光碟之後在室溫加以培養1小時，之後使用50 mM磷酸鹽 緩衝器（pH 7)加以清洗。 如此衍生之光碟之後用以驗定人體血液中之紅血球細胞 的呈現。 100 μΐ人體血液之樣本由正常志願者抽出。1〇〇 μΐ樣本 98815.doc -68 - 1249158 之可/刀里使用磷酸鹽-緩衝器鹼鹽（npBS")稀釋1〇倍。二種 ^卜10: 1系列稀釋相同地執行。-樣本之職升⑽可分 里乂及各系列稀釋之樣本之1〇 ^可分量·為分開和單獨置 放在4光碟驗定格點上。 口亥光碟在⑳閉潮濕器室内之氮氣下以室溫培養2小時。 該光碟之後以PBS清洗。 圖12為確定RBC免疫特定附著於光碟驗定格點的原子力 • atomic微觀影像。如量化分析所表示，RBC水平大小既定 為7.984 μπι，和紅血球細胞已知直徑吻合（8 ;此大小 和例子2所使用和觀察之膠乳球體的均勻3 直徑明顯地 不同。在紋道底部之上的RBC高度觀察為18 pm。 忒光碟以二元醇之5%溶液清洗，如下文在光碟機中加 以乾燥和讀取。 由CD Associates, Inc·(加州lrvine市）所製造用於光碟工 業品質控制使用的CD-R裝置為用以讀取該光碟。驅動裝 • 置的CD-R擺動尋軌系統（模式RSL1 〇〇)被加入之透鏡丨7修 正光學拾音器以在光碟上缺乏第一折射層時調整聚焦；轉 軸之尚度亦提咼。HF信號（RF或是四列總和）由RSLi〇〇中 的電子電路加以放大，以及該緩衝之HF信號輸入至數位示 波器。 圖13呈現代表性追縱，以X轴顯示時間而γ軸顯示四列 總和信號大小。圖13實際說明紅血球細胞直接可見地為 C D · R閱頃機H F #说南頻、南振幅事件；對喷乳動物細胞 大小之分析物而言，沒有膠乳球體或是其他外因發出信號 98815.doc -69- Ϊ249158 之一部份為所需以產生特定分析物信號。 由圖13之示波器追蹤亦明顯地是，1〇?基準線的差異為 雙重峰值。雖然紅血球細胞已知為具有特徵性雙凹形狀， 我們已經在例子2中觀察到此雙重峰值被用以報告分析物 之呈現。此雙重峰值出現為雷射遍歷紋道中之球體時，由 反射比中之再生變化形成。MagabeadsTM is a commercially available spheroidal latex magnetizable particle _ and a part of a different type of joint, such as here, "anti-human IgG or another reaction group that allows for custom conjugates." The disc is dried and its surface is visualized by light and atomic force microscopy (AFM). Figure 8 shows the determination of a specific analyte on a first surface by a portion of the IgG-specific analyte after addition to human blood and antibody-conjugated spheres. The optical image of the optical disc is captured by the microscopic view. Figure 9 and Figure 1 show the AFM image of a single latex sphere that is specifically attached to the disc. The magnification is slightly higher than the magnification used in Figure 8, and is summarized in the image. The size of the measurement was observed by afm. Figure 11 is an AFM image of the immune-specific attachment to the first surface to search for human IgG-specific discs and two latex spheres presented in the same field, and summarized by AFM during image capture. Observed measurement large / Jn 〇 Example 3 Electronic detection and characterization of human red blood cells on RBC-specific searchable immunoassay discs, single data layer, first surface, The image/positive embossed disc was fabricated according to Example 1. The gold surface of the disc was then derived as follows: 2 mg N-[6_(vitaminium amino)hexyl]_3,_(2,_pyridylthio)propyl Indoleamine (,, vitamin H-HPDP,,) (Pierce, Rockford, il; article number 98815.doc -67-1249158 97032461) may be dissolved in 2 ml of dimethylformamide. Each of the grid points in the intent of the intent is positioned at the same radius from the center of the disc, and the 10 μΐ vitamin H-HPDP solution is moved by a pipette. The disc is incubated at room temperature for 2 hours. And then washed with a 50 mM phosphate buffer (pH 7.) Next, 10 μΐ of streptavidin solution (Monobind, Costa Mesa, CA; product 96-001/MF; 2 mg/ml) was pipetted at the same At the assay site, the disc was incubated for 1 hour at room temperature and then washed with a 50 mM phosphate buffer. Monoclonal mouse anti-human glycophorin A antibody (Dako, Carpinteria, Calif.; product 113) was biotinylated as follows. The 100 μΐ component of the antibody and the α-vitamin of 0.1 mg Poly(ethylene glycol carbonate), (Bio-PEG-NHS,) (Shearwater, Inc., Huntsville, AL; PT-028-27) 100 μL of the sulphate buffer (pH 7) and allowed to react for 1 hour. Vitamin Η-conjugated anti-human glycophorin A was dialyzed against the same buffer (dialysis MWCO = 30,000) overnight. The dialyzed vitamin H-complex anti-human glycophorin A antibody was pipetted on a streptavidin-plating test spot on a disc and after the disc was incubated at room temperature for 1 hour, after which a 50 mM phosphate buffer was used. (pH 7) was washed. The disc thus derived is used to verify the appearance of red blood cells in human blood. A sample of 100 μΐ human blood was drawn from normal volunteers. 1〇〇 μΐ sample 98815.doc -68 - 1249158 can be diluted 1〇 times with phosphate-buffer base salt (npBS"). Two types of ^b 10:1 series dilutions were performed identically. - The sample's job-up (10) can be divided into 1 〇 ^ component of the sample of each series of dilutions and is placed separately and separately on the 4-disc test grid. The mouth-light disc was incubated at room temperature for 2 hours under nitrogen in a 20-closed humidifier chamber. The disc is then washed with PBS. Figure 12 is an atomic force microscopic image that determines the RBC immune specific attachment to the disc verification grid. As indicated by the quantitative analysis, the RBC level was determined to be 7.984 μm, which is consistent with the known diameter of the red blood cells (8; this size is significantly different from the uniform 3 diameter of the latex sphere used and observed in Example 2. Above the bottom of the groove The RBC height is observed at 18 pm. The disc is cleaned with a 5% solution of glycol, dried and read in an optical disc as follows. Fabricated by CD Associates, Inc. (Lrvine, Calif.) for disc industry quality The CD-R device used for control is used to read the CD. The CD-R wobble tracking system (mode RSL1 〇〇) of the drive device is added to the lens 丨7 to correct the optical pickup to lack the first refraction on the disc. Adjusting the focus at the time of the layer; the degree of rotation of the shaft is also improved. The HF signal (RF or four-column sum) is amplified by the electronic circuit in RSLi, and the buffered HF signal is input to the digital oscilloscope. Tracking, the time is displayed on the X-axis and the gamma-axis shows the sum of the four columns of the signal. Figure 13 shows that the red blood cells are directly visible as CD · R reading machine HF # said south frequency, south amplitude event; For cell size analytes, there is no latex sphere or other external cause signal 98815.doc -69- Ϊ249158 is required to generate a specific analyte signal. The oscilloscope trace from Figure 13 is also apparently 1 The difference between the baselines is a double peak. Although red blood cells are known to have a characteristic biconcave shape, we have observed in Example 2 that this double peak is used to report the presence of the analyte. This double peak appears as a laser. When the sphere in the track is traversed, it is formed by the regenerative change in the reflectance.

由圖13之示波器tracing另一觀察而言，容易見到地是在 發出信號事件各側上的基準線為穩定；也就是說，擺動紋 道（此處製造為inverse影像擺動紋道）之尋軌本身不造成 guad sum信號中的重大變化。From the other observation of the oscilloscope tracing of Figure 13, it is easy to see that the reference line on each side of the signalling event is stable; that is, the wobble track (made here as the inverse image wobble track) The rail itself does not cause significant changes in the guad sum signal.

tCD_R標準之光學閱讀機，無論正在光碟上的任何位 置讀取都料常數線性速度，修正轉軸速度以鎖定擺動紋 f之頻率。依據已知光碟線性速度和示波器追蹤上的增量 才下丁不波g哥軌上的各劃分可以顯示為相當於光碟上線 性距離U叫。如圖13顯示之示波器追縱所量測，四列總 和k號基準線中的差異因此岐為1()叫，近似尋軌紋道 方向之物件的非校正大小。 4物件貫際大小為較小。使用3㈣膠乳球體之閱讀機校 〆和不波器已經給予示波器峰值報告5 μ__大小， 際：件寬2 ’。此情形適當地在驗定光碟上的第一表面上 考量為1·5 μηι雷射聚焦直徑。 考ϊ介於量測和雷射直徑在光碟表面上發生實際大小之 門的2 μηι差異時，在圖13示波器尋軌捕獲的事件為四 和信號高頻、高振幅差異，報告…物件大小，和已二 988l5.doc -70-The tCD_R standard optical reader reads the constant linear velocity regardless of the position on the disc, and corrects the spindle speed to lock the frequency of the wobble. According to the known linear speed of the optical disc and the increment on the oscilloscope tracking, the divisions on the gamble can be displayed as equivalent to the linear distance U on the optical disc. As shown in the oscilloscope trace shown in Figure 13, the difference between the four columns and the reference line k is therefore 11 (), which approximates the non-corrected size of the object in the direction of the track. 4 The size of the object is small. Using a 3 (four) latex sphere reader calibration and non-wave device has given the oscilloscope a peak report of 5 μ__ size, width: 2 ’. This situation is appropriately considered to be a 1·5 μηι laser focusing diameter on the first surface on the verification optical disc. Considering the 2 μηι difference between the measurement and the actual diameter of the laser diameter on the surface of the disc, the event captured by the oscilloscope in Figure 13 is the sum of the high frequency and high amplitude of the signal, and the object size is reported. And already two 988l5.doc -70-

1249158 人體紅血球吻合。 圖14呈現由相同光碟上之分離紅血球細胞偵測發出信號 請事件另一示波器尋軌。雙相峰值為更顯著。圖15:7 為額外例子。 圖，由相同光碟上不同區域擷取之多數事件的數位重 疊’實際說明在-些免疫特錢著於光碟不同紅血球細胞 上大小和形狀量測之再生性。 例子4 最佳信號元素大小之計算 特定分析物信料以藉㈣整球體㈣元素㈣於尋軌 紋道之大小的大小而加以最佳化，如下文解釋。 圖30解釋fit至光碟362上之紋道的球體信號元素則大小 之計算，如此該信號元素在3點接合至該紋道： 底部’丨點在紋道各邊緣。在下列方程式中，_球體信號 素半徑，W為紋道寬度，以及h為紋道深度。1249158 Human red blood cells are anastomosed. Figure 14 shows the detection of a signal from a separate red blood cell on the same disc. The two-phase peak is more pronounced. Figure 15: 7 is an additional example. The figure, the digital overlap of most events taken from different areas on the same disc, actually illustrates the reproducibility of the size and shape measurements on the different red blood cells of the disc. Example 4 Calculation of the optimal signal element size The specific analyte tract is optimized by the size of the (4) element (4) element (4) in the orbital track, as explained below. Figure 30 illustrates the calculation of the size of the spherical signal element of the fit to the groove on the optical disk 362, such that the signal element is joined to the track at 3 points: the bottom '丨 is at each edge of the track. In the following equation, _ sphere signal radius, W is the groove width, and h is the groove depth.

藉由Pythag0rean定理，介於球體半徑和紋道寬度及深度 之間的關係為： XX '解答等於： ：(r-h)2+(^)2 0) 4h2 + w2 8h~ (2) ) 由於紋道深度較佳為"8,此處又為用以讀取光碟之光 波長’可以將半徑表示為.·· 的 又 (3) 98815.doc -71- 1249158 簡化此式等於： 一义 w2 Γ =--1-- 16 λ (4) 應用等式（4)，假使讀取光碟之光的波長為〇.65 μιη(亦即 650 nm ’該波長為用於DVD)，以及該紋道寬度為〇·8 pm(DVD之磁執節距），則球體信號元素半徑必須近似於 1.03 μηι 〇 例子5 最佳化於第一表面偵測之具有前向影像/正浮雕擺動紋道 之單一資料層光碟的製造 CD-R母片在CINRAM預訂而加以製作，基本上如例子1 所陳述’以作用為壓模而生產可尋軌、單一資料層、前向 影像/正浮雕擺動紋道之光碟。該母片為用以沖壓大約 5000片聚碳酸脂光碟，之後以金加以金屬化以及儲存於後 續使用。該光碟鑄造在ΕΧΙΜΡΟ S.R.O.(布拉格，捷克共和 國）執行。鑄模設定在圖41A-41I中陳述。 在例子1中製作之光碟（以及在例子和例子3使用以產生 呈現在圖8-1 8之資料），具有深度近似170 nm之紋道，報 告為由原子力微觀提供之大小總結的”垂直距離”，在圖^ 〇 中再生。相反地，此處製造的光碟，設計為紋道深度近似 第一表面，理論上最佳化為意圖之入射雷射波長78〇打历的 1/8。 圖31呈現其中之一光碟内徑之原子力微觀檢視所產生的 資料。大小總結報告深度近似1 〇〇 nm之紋道。圖32呈現其 中之一光碟外徑之原子力微觀檢視所產生的資料；如光碟 98815.doc -72- 1249158 製造之典型，該紋道深度稍大於外徑（此處為l〇l23 nm)， 以容納光碟在光軸方向之外部部分稍為增加的運行。圖33 呈現母片内徑之原子力微觀檢視所產生的類似資料，以及 圖34呈現母片外徑之原子力微觀檢視所產生的資料。 例子6 雷射-折射聚碳酸脂蓋板之製造 雷射折射聚碳酸脂蓋板，其中之一以上視圖顯示在圖35 以及在圖1 9以側面區段圖（組合為單一資料層特定分析物 光碟）更進一步示意，如以下步驟加以製造。 錄光碟壓模意圖用於標準CD製造，但是用於有效安裝 在鑄造機器上為太厚，被置放在標準壓模拋光器中。壓模 之負料表面為平滑性而拋光，產生具有二抛光面之大約 260 μηι _ 330 μηι厚的壓模。該壓模安裝在標準CD_R中且 a又疋调整為將聚碳酸脂蓋板既定為近似丨·丨7 mm厚，但是 另一方面大小等於標準12〇 mm光碟。 在例子5中製造之單一資料層光碟為大約12 +/_ 〇〇5爪爪 厚，蓋板為大約1.17 mm厚。二者加在一起建立近似2·4 mm厚的組件，外部最大實體厚度由紅皮書標準（所有層組 δ起來為1.1 _ 1.5 mm)提供。實驗性地，我們發現該光碟 組件之增加厚度不會呈現光學或是機械問題。呈現在圖40 中之資料為使用依據例子5所製造之光碟而獲得以及在讀 取之前以依據此例子製造的蓋板加以組合。該蓋板提供足 夠的輔助於聚焦以排除另一聚焦透鏡丨7增加至驅動裝置的 光學拾音器。 988I5.doc -73- 1249158 例子7 信號元素之高密度核酸順序驅動附接於可尋軌光碟 單一資料層、第一表面、前向影像/正浮雕擺動光碟為 依據例子5加以製造。該光碟金表面為如下之步驟加以衍 生。調處為在無塵室中以疊層流體方式加以執行。 6光碟中之各光碟上為置放15 pL streptavidin溶液（2 mg/mL)之單一點。該光碟為培養1小時，之後在蒸餾 H20(dH20)流中清洗。10 pL 2-氫硫基乙胺（137 pg/mL)之 可分量增加至各點以阻斷非特定之接合（該溶液已經在4°C 下儲存一段足以將溶液氧化的時間）。該光碟為培養3分 鐘，之後以dH20清洗。該光碟在使用之前不加以乾燥。 核酸探針以胺修正3’或是5’界柱合成以由Keystone實驗 室（加州Foster市）預訂，如下： 5，-TCGGGTGTACTCAC·胺-3，（SEQ ID NO ·· 1) 5，·胺-TCCAAGAAAGGACC-3f(SEQ ID NO : 2) 各探針之後如下文經由胺修正界柱獨立地共軛至維生素 Η。 維生素H-PEG-NHS之物料溶液（α _維生素Η，ω-Ν-脛基 丁二醯胺酯之聚（乙烯乙二醇）-碳酸酯）（MW=3400，阿拉巴 馬州 Huntsville市 Shearwater公司；貨品 PT-028-27)藉由溶 解固體在包含疊氮化物為防腐劑之磷酸鹽緩衝器之鹼鹽 (PBSAz ^ pH 7.45)^^:^>t^^23.5 nmol^L(4.7 mg維生 素 H-PEG-NHS在 58.82 μΙ〇。 該3’活性探針（SEQ ID NO : 1)(π3’探針”）為溶解在PBSAz 98815.doc -74- 1249158 至最終濃度為1 nmol/pL(473 nmol在473 pL)°3,探針之20 ML(20 mol)溶液之後加入至10 pL維生素H-PEG-NHS物料 溶液而等於最終核酸濃度為660 pmol/pL。同時，5’ aminated 探針（SEQ ID NO: 2)(”5,探針’，）為溶解在 PBSAz 至 最終濃度為2 nmol/pL(84 nmol在42 pL )，以及5’探針之 10pL(20 mol)溶液之後加入至1〇 μΐ^維生素H-PEG-NHS而 等於最終核酸濃度為1 nmol/pL。該溶液為分別在室溫（RT) 培養2小時。 該3’探針之後經由維生素Η—部份而共軛至單一分散相 超順磁性串珠s(直徑一致為2.8 μηι)表面。如上文準備的1〇 個連續1〇41^1:1〇稀釋3’1^〇^11>^16(1探針（1:1〇稀釋之66 pmol)之可分量加入至streptavidin-鍵層Dynabeads®(紐約州 Lake Success市Dynal公司；種類號碼為M-280號）1 mg可分 量（100 μι，10 mg/ml)。在室溫之10分鐘培養為介於第一 和第二探針溶液增加至串珠之間執行，5分鐘培養為介於 第二和第三之間以及2分鐘培養為介於其他增加之間執 行。 3·探針-共軛串珠之後以200 μί PBSAz和200 pL雜化作用 缓衝器加以清洗（923 mM Na2HP04，75 mM NaH2P04，1 mM EDTA，pH 7·34)二次。串珠使用磁性分離（Dynal)而由 清洗溶液量化地回復。串珠在1 mL雜化作用緩衝器中再次 懸浮。假設核酸完整共軛，呈現在此共軛串珠溶液之3’探 針平均濃度為198 fmol/pL。 5’ biotinylated探針（1 nmol/pL為20 pL)以雜化作用緩衝 988I5.doc -75- 1249158 器加以稀釋為1 mL(最終濃度為20 ρπιο1/μΙ〇。 目標核酸由Key stone實驗室綜合而預訂如下： 5，-GTGAGTACACCGGAATTGCCAGGACGACCGGGTCCTTT CTTGGA-3f(SEQ ID NO ： 3) 目標溶解在PBSAz為最終濃度100 μΜ。此後，4個1，000-倍（1 μΐ^比1 mL)系列稀釋以雜化作用緩衝器加以執行，形 成目標測試溶液為濃度1〇〇 nM、100 pM、100 fM和1〇〇 aM 〇 6個流體相位之雜化作用反應以平行溶液加以設定。在 各反應中，2 pL串珠-共軛之3,探針（396 f莫耳核酸）以及1 μί biotinylated 5,探針（20 p莫耳）以目標 DNA(SEQ ID NO : 3)加以培養如下： 反應1 1 μί 100 nM目標=100 f莫耳目標 反應2 1 pL 100 pM目標=100 a莫耳目標 反應3 1 μί 100 fM目標=100 z莫耳目標 反應4 1 μι 100 aM目標=100 y莫耳目標 反應5 1 pL雜化作用緩衝器（控制） 反應6 1 gL水（控制） 各試管在室溫時在300 RPM之攪拌器上培養2小時。 在雜化作用完成之後，6雜化作用反應中之各反應以1〇〇 μί PBSAz加以清洗二次，而串珠由清洗溶液量化地回 復，串珠在10 pL PBSAz中再次懸浮。對6反應中之各反應 而言，2 pL之串珠懸浮之後應用至分離之一光碟上的驗定 點，形成目標總量如下： 98815.doc -76- 1249158 光碟1 20 f莫耳（2〇χ1(Γ15莫耳）目標 光碟2 20 a莫耳（2〇χ1〇·18莫耳）目標 光碟3 20 2莫耳（2(^10·21莫耳）目標 光碟4 20 y莫耳（2〇χ1(Γ24莫耳）目標 光碟5 〇(雜化作用緩衝器控制） 光碟6 〇(水控制） 串珠在光碟上培養1〇分鐘，之後以水流清洗。光碟加以 乾燥，之後使用輕顯微鏡加以視覺化。 圖3 6在視覺化時間示意該驗定格點。直接附著於可尋執 光碟之金表面的是streptavidin之鍍層，藉由韋德瓦斯力和 形成於streptavidin蛋白質之自由sulfyhydryl和光碟金表面 之間的&化金黏合劑加以結合。該streptavidin捕捉5,探針 之維生素H-部份。該5丨探針依次由邮甘酸之伽議_With the Pythag0rean theorem, the relationship between the radius of the sphere and the width and depth of the groove is: XX 'The solution is equal to: :(rh)2+(^)2 0) 4h2 + w2 8h~ (2) ) Due to the groove The depth is preferably "8, here is the wavelength of the light used to read the disc', and the radius can be expressed as ..·(3) 98815.doc -71- 1249158 Simplified this formula is equal to: Yiyi w2 Γ =--1-- 16 λ (4) Apply equation (4), if the wavelength of the light reading the disc is 〇.65 μιη (that is, 650 nm 'this wavelength is for DVD), and the width of the groove For 〇·8 pm (the magnetic pitch of the DVD), the radius of the sphere signal element must be approximately 1.03 μηι 〇 Example 5 Optimized for the first surface detection of a single image with forward image/positive embossed wobble track Fabrication of a layered disc CD-R master is produced at CINRAM and is basically as described in Example 1 'Production of a trackable, single data layer, forward image/positive embossed wobble track by acting as a stamper . The master is used to punch approximately 5,000 sheets of polycarbonate disc, which is then metallized with gold and stored for subsequent use. The disc was cast in ΕΧΙΜΡΟ S.R.O. (Prague, Czech Republic). The mold settings are set forth in Figures 41A-41I. The discs produced in Example 1 (and used in Examples and Example 3 to produce the data presented in Figure 8-1 8) have grooves with a depth of approximately 170 nm, reported as "vertical distances" summarized by the size provided by the atomic force microscopic ", reproduced in Figure ^ 。. Conversely, the disc manufactured here is designed to have a groove depth approximating the first surface and is theoretically optimized to 1/8 of the intended incident laser wavelength of 78 〇. Figure 31 presents data generated by microscopic examination of the atomic force of one of the inner diameters of the disc. The size summary report is approximately 1 〇〇 nm in depth. Figure 32 shows the data generated by the atomic force microscopic examination of one of the outer diameters of the optical disc; as typical as manufactured by the optical disc 98815.doc -72-1249158, the groove depth is slightly larger than the outer diameter (here, l〇l23 nm), A slightly increased operation of the outer portion of the optical disc in the direction of the optical axis is accommodated. Figure 33 shows similar data from an atomic force microscopic examination of the inner diameter of the master, and Figure 34 shows the data generated by microscopic microscopic examination of the outer diameter of the master. Example 6 Laser-Reflective Polycarbonate Cover Plate Manufactured Laser Refractive Polycarbonate Cover Plate, one of which is shown in Figure 35 and in Figure 19 in a side section view (combined as a single data layer specific analyte) The disc) is further illustrated and manufactured as follows. The disc stamper is intended for standard CD manufacturing, but is used for effective mounting on casting machines that are too thick and placed in a standard stamper polisher. The negative surface of the stamper is smoothed and polished to produce a stamper having a thickness of about 260 μηι _ 330 μηι thick with two polished faces. The stamper is mounted in the standard CD_R and a is adjusted to set the polycarbonate cover to approximately 丨·丨 7 mm thick, but on the other hand the size is equal to the standard 12 〇 mm disc. The single data layer disc manufactured in Example 5 was approximately 12 +/_ 〇〇 5 claw thick and the cover was approximately 1.17 mm thick. Together, the two components are approximately 2·4 mm thick, and the maximum external solid thickness is provided by the Red Book standard (all layers are δ 1.1 _ 1.5 mm). Experimentally, we have found that the increased thickness of the disc assembly does not present optical or mechanical problems. The data presented in Figure 40 was obtained using the discs manufactured according to Example 5 and combined with a cover plate made in accordance with this example prior to reading. The cover plate provides sufficient optical pickup to assist in focusing to exclude the addition of another focusing lens 丨7 to the drive unit. 988I5.doc -73- 1249158 Example 7 High-density nucleic acid sequential driving of signal elements attached to a track-receivable disc A single data layer, a first surface, a forward image/positive embossed oscillating disc is manufactured according to Example 5. The disc gold surface is derived as follows. The transfer is performed in a layered fluid manner in a clean room. A single point of 15 pL of streptavidin solution (2 mg/mL) was placed on each of the 6 discs. The disc was incubated for 1 hour and then rinsed in a stream of distilled H20 (dH20). A 10 pL 2-hydrothioethylamine (137 pg/mL) component can be added to each point to block non-specific bonding (the solution has been stored at 4 ° C for a period of time sufficient to oxidize the solution). The disc was incubated for 3 minutes and then washed with dH20. The disc is not dried before use. The nucleic acid probe was modified with an amine 3' or 5' column to be ordered by Keystone Laboratories (Foster, Calif.) as follows: 5,-TCGGGTGTACTCAC·amine-3, (SEQ ID NO ··1) 5,·amine -TCCAAGAAAGGACC-3f (SEQ ID NO: 2) Each probe was then independently conjugated to vitamin oxime via an amine-corrected binding column as follows. Vitamin H-PEG-NHS material solution (α _ Η Η, ω-Ν-胫 丁 丁 醯 之 (polyethylene glycol)-carbonate) (MW = 3400, Shearwater, Huntsville, Alabama Company; product PT-028-27) by dissolving solid in a phosphate buffer containing azide as a preservative (PBSAz ^ pH 7.45) ^^:^>t^^23.5 nmol^L (4.7 The mg of vitamin H-PEG-NHS is at 58.82 μΙ〇. The 3' active probe (SEQ ID NO: 1) (π3' probe) is dissolved in PBSAz 98815.doc -74-1249158 to a final concentration of 1 nmol/ pL (473 nmol at 473 pL) °3, probe 20 ML (20 mol) solution was added to 10 pL of vitamin H-PEG-NHS material solution equal to the final nucleic acid concentration of 660 pmol/pL. Meanwhile, 5' aminated Probe (SEQ ID NO: 2) ("5, probe',) was dissolved in PBSAz to a final concentration of 2 nmol/pL (84 nmol at 42 pL), and a 5' probe of 10 pL (20 mol) solution Thereafter, it was added to 1 〇μΐ^vitamin H-PEG-NHS to be equal to a final nucleic acid concentration of 1 nmol/pL. The solution was incubated at room temperature (RT) for 2 hours. The 3' probe was then passed through a vitamin Η-part and Yoke to a single dispersed phase superparamagnetic bead s (consistent diameter of 2.8 μηι) surface. As prepared above, 1 连续 continuous 1〇41^1:1〇 dilution 3'1^〇^11>^16(1 probe (6 pm diluted 66 pmol) can be added to the streptavidin-bond layer Dynabeads® (Dynal Company, Lake Success, NY; type number M-280) 1 mg component (100 μιη, 10 mg/ml) The incubation at room temperature for 10 minutes is performed between the addition of the first and second probe solutions to the beading, the incubation for 5 minutes is between the second and third, and the incubation for 2 minutes is between the other increases. 3. Perform probes. 3. Probe-conjugated beads were washed twice with 200 μί PBSAz and 200 pL hybrid buffer (923 mM Na2HP04, 75 mM NaH2P04, 1 mM EDTA, pH 7.34). The magnetic separation (Dynal) was quantitatively recovered by the cleaning solution. The beads were resuspended in a 1 mL hybrid buffer. Assuming the nucleic acid was fully conjugated, the average concentration of the 3' probe present in the conjugated bead solution was 198 fmol/ pL. The 5' biotinylated probe (20 pL at 1 nmol/pL) was diluted to 1 mL with a hybridization buffer 988I5.doc -75-1249158 (final concentration 20 ρπιο1/μΙ〇. Target nucleic acid synthesized by Keystone Laboratory) The booking is as follows: 5,-GTGAGTACACCGGAATTGCCAGGACGACCGGGTCCTTT CTTGGA-3f (SEQ ID NO: 3) The target is dissolved in PBSAz to a final concentration of 100 μΜ. Thereafter, four 1,000-fold (1 μΐ^ to 1 mL) serial dilutions are used for hybridization. The action buffer is executed to form a target test solution with a concentration of 1 〇〇 nM, 100 pM, 100 fM and 1 〇〇 aM 〇 6 fluid phases of the hybridization reaction set in parallel solutions. In each reaction, 2 pL Bead-conjugated 3, probe (396 f-mole nucleic acid) and 1 μί biotinylated 5, probe (20 pmol) were incubated with the target DNA (SEQ ID NO: 3) as follows: Reaction 1 1 μί 100 nM Target = 100 f Mohr target response 2 1 pL 100 pM target = 100 a Mo target response 3 1 μί 100 fM target = 100 z Mohr target response 4 1 μι 100 aM target = 100 y Mohr target response 5 1 pL Hybrid Buffer (Control) Reaction 6 1 gL Water (Control) Incubate for 2 hours at room temperature on a stirrer at 300 RPM. After the completion of the hybridization, each reaction in the 6 hybridization reaction was washed twice with 1 μμί PBSAz, and the beads were quantitatively recovered from the washing solution. The beads were resuspended in 10 pL PBSAz. For each reaction in the 6 reaction, 2 pL of bead suspension was applied to the verification point on one of the discs to form the target total amount as follows: 98815.doc -76- 1249158 Disc 1 20 f Mo Er (2〇χ1 (Γ15 Moer) Target Disc 2 20 a Mo (2〇χ1〇·18 Molar) Target Disc 3 20 2 Mo Er (2 (^10·21 Moel) Target Disc 4 20 y Moule (2〇χ1 (Γ24mol) Target Disc 5 〇 (Hybrid Buffer Control) Disc 6 〇 (Water Control) Beads are incubated on the disc for 1 minute, then washed with water. Dry and then visualize using a light microscope. Figure 3 6 shows the verification grid point at the visualization time. The gold surface directly attached to the rewritable disc is the streptavidin coating, which is formed by the Wedwars force and the streptavidin protein. Free sulfyhydryl and disc gold watch Between & binder be combined and gold. The streptavidin captures 5, the vitamin H- moiety of the probe. The 5丨 probe is in turn by the postal acid _

Crick補充在目標3,端捕獲目標核順序。該目標依次在5，端 經由Μ核甘酸之Wats〇n_Crick補充捕獲3，端，因此繫鏈 Dynabead®至該光碟。 圖37呈現由驗定光碟μ分別採取之光微觀影像，各光 碟放大率為2 :附著球體和擺動紋道可清楚地以較高放大 率面板之所有3影像看見。增加數目之附著串珠可清楚地 以增加數量之㈣目標看見，而光碟3(圖37〇顯示球體以 20十兆億*之-莫耳（2〇χ1〇-21莫耳；ΐ2χΐ〇3莫耳）核酸目標 補充-驅動附著至光碟表面，而光碟2(圖37Β)顯示球體以 百億億分之-莫耳（2〇χ1〇·18莫耳；12χΐ〇6莫耳）核酸目標補 充-驅動附著至光碟表面，而光碟U圖37Α)顯示球體以20 988l5.doc -77- 1249158 千萬億分之-莫耳⑽心莫耳；12xig9莫耳）核酸目標補 充:’駆動附著至光碟表面。沒有串珠在其中之一控制光碟 (沒有顯示）表面上觀察到。 此處所提到之所有專利、專利公告和其他發表之文獻假 使各文獻已經單獨地和特定地以提及的方式併入本文，則 在此以提及的方式全部併入本文中。而說明之本發明較佳 解釋性具體實施例對熟知相關技藝之人士而言將明顯的是 籲 種不同麦化和修正可以由此以不脫離本發明範缚之下加 以製作，以及意圖在申請專利範圍中涵蓋所有落入本發明 精神和範疇之内的此種變化和修正。 【圖式簡單說明】 本發明上述和其他目的及優點依據與附圖有關聯之下列 詳細說明的考量而變得明顯，該圖不以一定比例描繪，其 中類似文字在整個附圖中引用為類似元件，以及其中： 圖1顯示單層CD類別光碟和閱讀機，而圖1A呈現定向為 » —CD光碟之㈣機光學拾音器的側視圖，該光碟以側 面橫斷面圖顯示，而雷射光路徑以線指示；而圖1B以較大 放大率顯示相同定向之光碟側面橫斷面圖；以及圖lc顯示 具有擺動紋道之CD-R光碟的透視圖； 圖2A和圖2B分別顯示典型3_射束拾音器相對於光碟磁執 之射束位置，以及使用3-射束於尋執、聚焦和讀取之典範 光碟偵測器與相關電子； 圖3顯示用於建立單一資料層光碟，其中圖3入顯示標準 電~和基體化處理，圖3 B顯示如本發明之修正處理，其中 988l5.doc -78- 1249158 母片使用為壓模’建立如本發明之前向影像、正浮雕倒置 光碟；圖3C顯示在圖3入中沖屡之傳統cd_^碟基板以反 射層和防護層完成，相對於訊問雷射加以定向；圖3D顯示 在圖3B中屋印之前向影像/正浮雕倒置光碟以反射層和非 知作、構凡成，相對於傳統訊問雷射加以定向·圖π顯示 本發明藉由在標準電鑄和基體化處理中使用反向影像主片 光碟而獲得之前向影像/負浮雕倒置光碟； • ® 4顯示應用於以側面橫斷面圖顯示之傳統單-層CD·類 別光碟之空間入射、雷射最接近第一表面的結構，實際說 明至閲讀機光學拾音器以及由閱讀機光學拾音器產生之光 學路徑的中斷； 圖5實際說明藉由將傳統CD_類別光碟加以反向而發生之 一類型反轉，而圖5A和圖5B以側面橫斷面圖實際說明實 體結構之反轉，雷射遠端結構導致雷射接近以及雷射接近 結構成為雷射遠端，而圖5C和圖5D實際說明尋執結構反 ® 轉影像，此處為螺旋； 圖6為定位於讀取以側面橫斷面圖顯示之本發明單一 層、第-表面、可尋執特定分析物光碟之光學拾音器側視 圖而雷射光學路徑以線指#，而圖6A實際說明在另外增 加聚焦透鏡至光學拾音器之前的焦點，而圖6B實際說明另 外增加¾^焦透鏡至光學拾音器時焦點的改變； 圖7示意分子成分，建立在本發明第一表面、可尋執特 定分析物光碟之反射表面上的1§(}_特定免疫分析，而圖7A 顯示樣本增加之前的驗定格點而圖7B顯示由人體血液樣本 98815.doc -79- 1249158 中之IgG調解的膠乳球體之牯定备 股 < 符疋免疫附者，接著增加人體 血液樣本以及此後另外藉由辦 广稽由i曰加抗_IgG共軛膠乳球體而，， 發展”； 圖8為應用人體血液和抗體共輛膠乳球體之後由部分 阶特定第-表面特定分析物可尋軌驗定光碟之光微觀檢 視捕獲的視頻影像； 圖9和圖1G為特定免疫附著至第―表面可尋軌人體-砂Crick added the target kernel order at target 3. The target is then supplemented at the 5th end by the Wats〇n_Crick of the nucleoside, which captures the 3rd end, thus tethering the Dynabead® to the disc. Figure 37 presents a microscopic image of the light taken by the verification optical disc μ, each having a magnification of 2: the attached sphere and the wobbled track are clearly visible to all 3 images of the higher magnification panel. An increased number of attached beads can be clearly seen with an increased number of (4) targets, while Disc 3 (Figure 37〇 shows a sphere with 20 billion trillion*-mole (2〇χ1〇-21mol; ΐ2χΐ〇3mol) ) The nucleic acid target is supplemented - the drive attaches to the surface of the disc, while the disc 2 (Fig. 37A) shows the sphere with a multimillion-mole-mole (2〇χ1〇·18 mol; 12χΐ〇6 mol) nucleic acid target supplement-drive Attached to the surface of the disc, and the disc U Figure 37 Α) shows the sphere with 20 988l5.doc -77-1249158 mpg-mole (10) heart moth; 12xig9 mol) nucleic acid target supplement: 'tilt attached to the disc surface. No beading was observed on the surface of one of the control discs (not shown). All of the patents, patent publications, and other published documents referred to herein are hereby incorporated by reference in their entirety in their entireties as if they are incorporated herein by reference. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT OF THE INVENTION The present invention will be apparent to those skilled in the art, and it is obvious that various modifications and modifications can be made without departing from the scope of the invention, and All such variations and modifications are intended to be included within the scope of the invention. BRIEF DESCRIPTION OF THE DRAWINGS The above and other objects and advantages of the present invention will become more apparent from the following detailed description of the accompanying drawings in which FIG. Components, and wherein: Figure 1 shows a single layer CD category disc and reader, while Figure 1A shows a side view of a (4) machine optical pickup oriented to a -CD disc, the disc being shown in a side cross-sectional view, and the laser light path Figure 1B shows a side view of the same orientation of the disc with a larger magnification; and Figure lc shows a perspective view of the CD-R disc with the wobbled track; Figures 2A and 2B show a typical 3_ The beam position of the beam pickup relative to the magnet, and the exemplary optical disc detector and associated electronics using 3-beam for seek, focus and read; Figure 3 shows the setup for a single data layer disc, where 3 shows the standard electrical and substrate processing, and FIG. 3B shows the correction processing according to the present invention, wherein the 988l5.doc -78-1249158 master is used as a stamper' to create a front image, positive relief as in the present invention. The optical disc is shown in Fig. 3C. The conventional cd_^ disc substrate in Fig. 3 is completed with a reflective layer and a protective layer, and is oriented relative to the interrogating laser. Fig. 3D shows the image/positive relief before the eave print in Fig. 3B. The inverted optical disc is oriented with a reflective layer and an unknown structure, and is oriented relative to a conventional interrogation laser. Figure π shows the present invention by using a reverse-image main-disc optical disc in standard electroforming and substrate processing. Image/negative embossed inverted disc; • ® 4 display applied to the space of the traditional single-layer CD·category disc displayed in side cross-section, the structure of the laser closest to the first surface, the actual description to the reader optical pickup And the interruption of the optical path produced by the optical pickup of the reader; Figure 5 actually illustrates one type of inversion by reversing the conventional CD_class disc, while Figures 5A and 5B are actually illustrated in a side cross-sectional view. Inversion of the physical structure, the laser distal structure causes the laser to approach and the laser close to the structure to become the far end of the laser, while Figures 5C and 5D actually illustrate the reverse structure of the seek structure, here the spiral; 6 is a side view of the optical pickup positioned to read a single layer, a first surface, a searchable optical disc of the present invention displayed in a side cross-sectional view, and the laser optical path is a line finger #, and FIG. 6A actually illustrates In addition to additionally increasing the focus of the focusing lens to the optical pickup, while FIG. 6B actually illustrates the change in focus when additionally adding a 3⁄4^ focus lens to the optical pickup; FIG. 7 illustrates the molecular composition, established on the first surface of the present invention, for searchable specific analysis 1 § (} _ specific immunoassay on the reflective surface of the disc, while Figure 7A shows the assay grid before the sample is increased and Figure 7B shows the latex sphere mediated by the IgG in human blood sample 98815.doc -79-1249158牯定备股< 疋 疋 疋 疋 , , , 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋After a total of latex spheres, a partial-specific specific surface-surface specific analyte can be used to determine the video image captured by the light microscopic examination of the optical disc; Figure 9 and Figure 1G are specific immune attachments to the first table. May seek human - sand

特定光碟之單-膠乳球體的原子力微觀（afm)影像，使用 較圖8稍高的放大率，具有在影像榻取期間由綱所觀察 之總結定量大小； 圖η為特定免疫附著至第一表面可尋軌人體_igG特定光 碟之二膠乳球體的原子力微觀（AFM)影像以及呈現在相同 AFM領域中’具有在影像操取期間由娜所觀察之總結定 量大小；The atomic force microscopic (afm) image of a single-latex sphere of a particular disc, using a slightly higher magnification than that of Figure 8, with a summary quantitative size observed by the scheme during image capture; Figure η is the specific immunization attached to the first surface The atomic force microscopic (AFM) image of the sigma sphere of the _igG-specific disc can be tracked and presented in the same AFM field as having a summary quantitative size observed by Na during the image manipulation;

圖12為AFM影像以及特定免疫附著至第一表面可尋軌人 體-RBC·特定光碟的紅血球⑽c)定量大小總結； 夺圖13為顯示由光碟閱讀機在特定免疫附著至第一表面可 尋執人體.RBC·特疋光碟之RBC上可尋軌掃描所獲得册（四 列總和)信號之特定分析物擾動的數位示波器追蹤，X軸顯 不時間而γ軸顯示四列總和信號大小； 圖14-17呈現顯示由光碟閱讀機在特定免疫附著至第一 表面可哥軌人體-!^^特定光碟之一些分離上可尋軌 掃描所獲得HF(四列總和)信號之特定分析物擾動的數位示 波器追縱’ X軸顯示時間而Y軸顯示四列總和信號大小； 98815.doc -80- 1249158 圖18由相同光碟擷取之多數事件的數位重疊，實際說明 在些特疋免疫附者至光碟之不同RB C大小和形狀量測之 再生性； 圖19為本發明以非整合雷射接近、雷射折射蓋板之單一 層可哥軌特定分析物光碟的側面橫斷面圖； 圖20顯不本發明單一層可尋執特定分析物光碟的較佳具 體實施例，以反射全像編碼之光碟操作結構反射與在光碟 第一表面上配置之非操作結構共焦的平面中之光碟尋轨結 構的影像； 一圖21顯线據區域心請性速度（ZCLv)標準建造之光碟 實體組織的簡化上側視圖； 圖22顯示圖21之ZCLV来碟甘士 九碟其中之一扇區的放大透視 圖23顯示配置在ZCLV第一砉而姑〜、 表面特疋分析物光碟扇區之Figure 12 is a quantitative summary of AFM images and red blood cells (10) c) with specific immune attachment to the first surface trackable human body - RBC · specific CD; Figure 13 is a display of the specific immune attachment to the first surface by the optical disk reader The RBC of the human body. RBC·Special Disc is available on the RBC for tracking search. The four-column sum signal is oscillating by the digital oscilloscope. The X-axis shows no time and the γ-axis shows the four-column sum signal size. -17 presenting the digits of the specific analyte perturbation of the HF (four-column sum) signal obtained by the optical disc reader in a particular immune attachment to the first surface. The oscilloscope traces the 'X-axis display time and the Y-axis shows the four-column sum signal size; 98815.doc -80- 1249158 Figure 18 The digital overlap of most events captured by the same disc, the actual description of the special immune attached to the disc Figure 19 is a side cross-sectional view of a single layer of a Kobe-specific analyte optical disc with a non-integrated laser approaching, laser-refractive cover plate of the present invention; Figure 20 A preferred embodiment of the present invention, in which a single layer can be searched for a particular analyte optical disc, reflects the optical disc in a plane that reflects the holographically encoded optical disc operating structure and is in focus with the non-operating structure disposed on the first surface of the optical disc. Figure 21 shows a simplified upper side view of the physical organization of the disc constructed according to the regional heart rate (ZCLv) standard; Figure 22 shows an enlarged perspective view of one of the sectors of the ZCLV of Figure 21. Display configuration in ZCLV first and then ~, surface special analysis of the optical disc sector

擺動紋面和紋道區域上的非操作結構； 圖24顯示典型雙層DVD袼式本泄 一 俗式先碟之側面橫斷面圖； 圖2 5顯示可組合雙資料居姓— 疋分析物*定光碟的放大 側面透視圖； 之雙層特定分析物驗定光碟具 強結構之雙層特定分析物驗定 圖26顯示包含内部通道 體實施例的側面橫斷面圖 圖27顯示以内部驗定_加 光碟的側面橫斷面圖； 驗定光碟的側面 圖28本發明另一顯示雙層特定分析物 橫斷面圖； 98815.doc •81 - 1249158 圖29顯示雙層特定分析物光碟的另一具體實施例； 圖30顯示喃合在光碟紋道中球面發出信號一部份之侧面 橫斷面圖，榡示各種尺寸大小； 圖3 1呈現本發明單一資料層光碟内徑之afm檢視產生的 資料，該光碟更進一步最佳化於相對圖8_圖12的第一表面 摘測’以及具有大約近似1 〇〇 nm之報告紋道深度； 圖32呈現本發明單一資料層光碟外徑之AFM檢視產生的 資料’該光碟更進一步最佳化於相對圖8_圖12的第一表面 摘測’以及具有大約近似101 nm之報告紋道深度； 圖3 3呈現用以沖壓在圖3 1和圖3 2量測之光碟的，，母片，，内 徑之AFM檢視產生的資料； 圖3 4呈現用以沖壓在圖3 1和圖3 2量測之光碟的，，母片，，外 徑之AFM檢視產生的資料； 圖35為聚碳酸脂雷射折射蓋板的上透視圖，和圖31和圖 32之光碟一起使用已產生如圖4〇所示之資料； 圖36示意建立在本發明可尋執光碟上的核酸基礎之特定 分析物驗定格點，其中單一2·8 μηι球體特定附著至光碟表 面為藉由核酸順序補充加以驅動； 圖37呈現3光碟之光微觀影像，各影像為2放大率，利用 圖36所示之驗定幾何加以建立，圖37Α顯示球體之核酸順 序補充驅動附著至20千萬億分之一莫耳（2〇χ1〇-ΐ5莫耳）目標 核酸；圖37Β顯示球體之核酸順序補充驅動附著至2〇百億 億分之一莫耳（20x1ο·〗8莫耳）目標核酸’·圖37C顯示球體之 核酸順序補充驅動附著至2〇十兆億分之一莫耳 988I5.doc -82- 1249158 耳）目標核酸， 圖38為在等於圖31和圖32中以AFM量測的那些光碟雷射 隶接近表面大小的300 X放大率擷取之二維合成光微觀影 像，2·8 μπι球體為靜電附接於金屬化表面以及實質上手動 地沿紋道對齊； 圖39為如圖38所示之相同光碟的較高放大率部分； 圖40顯示沿著通過圖39顯示之光碟區域中的單一磁軌之 HF信號報告的電反應；以及 、圖41呈現例子5中說明之光碟製造所使用鑄楔的設定，Non-operating structure on the oscillating surface and track area; Figure 24 shows a side cross-sectional view of a typical double-layer DVD 本 泄 一 ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; *An enlarged side perspective view of the fixed disc; a two-layer specific analyte verification of the disc with a strong structure of the two-layer specific analyte verification Figure 26 shows a side cross-sectional view of the embodiment containing the internal channel body Figure 27 shows the internal test Side cross-sectional view of a fixed-disc disc; side view of the verification optical disc 28 Another cross-sectional view showing a bi-layer specific analyte of the present invention; 98815.doc •81 - 1249158 Figure 29 shows a bi-layer specific analyte optical disc Another embodiment is shown in FIG. 30; FIG. 30 is a side cross-sectional view showing a part of a spherical signal emitted from a track in a disc track, showing various sizes; FIG. 31 shows the afm inspection of the inner diameter of the single data layer of the present invention. The disc is further optimized for the first surface pick-up with respect to Figures 8-12 and the reported track depth of approximately 1 〇〇 nm; Figure 32 shows the outer diameter of the single data layer disc of the present invention. AFM inspection produced The data 'The disc is further optimized for the first surface picking with respect to Figures 8 - 12" and has a reported track depth of approximately approximately 101 nm; Figure 3 3 is presented for stamping in Figures 31 and 3 2 Measurement of the disc, master, and AFM inspection of the inner diameter; Figure 3 4 shows the disc used to stamp the discs measured in Figure 31 and Figure 3, the master, the outer diameter Figure 35 is a top perspective view of a polycarbonate laser refracting cover plate, and the use of the optical disks of Figures 31 and 32 has produced the data as shown in Figure 4A; Figure 36 illustrates the construction of the present invention. A specific analyte checkpoint can be found on the basis of the nucleic acid on the disc, wherein a single 2·8 μηι sphere is specifically attached to the surface of the disc to be driven by the nucleic acid sequence supplement; FIG. 37 presents a microscopic image of the 3 discs, each image being 2 magnification, established using the assay geometry shown in Figure 36, Figure 37 shows that the nucleic acid sequence of the sphere is supplementally driven to attach to 20 ppm of a molar (2〇χ1〇-ΐ5 mole) target nucleic acid; 37Β shows the nucleic acid sequence of the sphere supplemental drive attached to 2〇100 One hundred millionth of a mole (20x1ο·8m) target nucleic acid'· Figure 37C shows the nucleic acid sequence of the sphere supplemental drive attached to 2〇10 trillionth of a molar 988I5.doc -82-1249158 ear) target Nucleic acid, Fig. 38 is a two-dimensional synthetic light microscopic image taken at 300 X magnification which is equal to the surface size of those optical discs measured by AFM in Fig. 31 and Fig. 32, and the 2·8 μπι sphere is electrostatically attached. Aligned on the metallized surface and substantially manually along the track; FIG. 39 is a higher magnification portion of the same optical disk as shown in FIG. 38; and FIG. 40 shows a single magnetic track in the optical disk region shown through FIG. The electrical response of the HF signal report; and FIG. 41 shows the setting of the casting wedge used in the manufacture of the optical disk described in Example 5,

八^不隹園31、32、37、38和39。 【主要元件符號說明】 10 描繪光學拾音器（接物組件） 11 標準CD-類別光碟 12-U 透鏡 13 物鏡 15 光束***器 16 散光元件 18 偵測器 19 雷射源 20 四分波片 70 維生素H-HPDP 72 Streptavidin 74 維生素Η—部份 76 Biotinylated goat 抗-人體 igG 988l5.doc -83- 1249158 78 IgG 79 固定抗-人體IgG Magabeads 112 透明基板 114 反射層 115 資訊層（資料層） 116 防護層 118 擺動紋道 132 基板 134 反射層 136 操作結構 194 全像 340 光碟 342 中央部分 346 鄰近通道 350 光碟 352 全像 354 影像平面 356 通道 358 間隔層 505 步驟 510 主片 515 步驟 520 父片 525 步驟 98815.doc -84- 1249158 530 母片 535 步驟 540 子片 625 步驟 A 平面 Au 光碟金表面 B 平面Eight ^ 不隹园 31, 32, 37, 38 and 39. [Main component symbol description] 10 Depicting optical pickup (splicing assembly) 11 Standard CD-class disc 12-U lens 13 objective lens 15 beam splitter 16 astigmatism element 18 detector 19 laser source 20 quarter-wave plate 70 vitamin H -HPDP 72 Streptavidin 74 Vitamin Η - Part 76 Biotinylated goat Anti-human igG 988l5.doc -83- 1249158 78 IgG 79 Immobilized anti-human IgG Magabeads 112 Transparent substrate 114 Reflective layer 115 Information layer (data layer) 116 Protective layer 118 Swing Track 132 Substrate 134 Reflective Layer 136 Operating Structure 194 Full Image 340 Disc 342 Central Section 346 Adjacent Channel 350 Disc 352 Full Image 354 Image Plane 356 Channel 358 Spacer 505 Step 510 Main Film 515 Step 520 Parent Sheet 525 Step 98815.doc -84- 1249158 530 Master 535 Step 540 Sub-Chip 625 Step A Plane Au Disc Gold Surface B Plane

98815.doc -85-98815.doc -85-

Claims (1)

1249158 公告本 十、申請專利範圍： ——— 1 · 一種光碟，其包括： 基板其具有一圖案化表面及一額外表面，該圖案 化表面提供該光碟之一資訊層； 複數個化學反應特徵，其係配置於該光碟之一空間入 射表面上；以及1249158 Announcement 10, the scope of the patent application: ——— 1 · A disc comprising: a substrate having a patterned surface and an additional surface, the patterned surface providing an information layer of the optical disc; a plurality of chemical reaction features, It is disposed on a spatial incident surface of the optical disc; 一操作結構，其係包含於該資㈣巾，該操作結構在 當該基板圖案化表面較該基板額外表面靠近一雷射時係 可由該雷射尋軌的； 其中-《多個肖等複數個化學反應特徵係配 操作結構可同時讀取的。 u 2·如4求項1之光碟’其中該操作結構係可尋執地配置為 一正浮雕前向影像。 ，、，、 3 · 如請求項彳 止 — 先碟，，、中該操作結構係可尋執地配置於 4 μ貝衹層離該基板額外表面最遠之表面。 一 ^ 、之光碟，其中该操作結構係可尋執地配置為 貞〉予雕前向影像。 5·如請求項] 今資1 s 先碟，其中該操作結構係可尋軌地配置於 :现層離該基板額外表面最近之表面。 7·如請求^之光碟’其中該操作結構包括-擺動紋道。 該操作〜‘之光碟’其中該等複數個化學反應特徵係與 、、、"構共焦配置。 8· 如請求垣^ 層，且复φ,光碟，其中該資訊層進—步包括-反射 、該等複數個化學反應特徵在當該基板圖案化 988I5.doc 1249158 彡基板額外表面接近該雷射時係配置為較該反射 層接近該雷射。 月长項1之光碟，其中該資訊層進一步包括一反射 層。 ，員9之光碟’丨中該等複數個化學反應特徵係配 置於該反射層之一空間入射表面上。 H· —種可尋執光碟，其包括·· 貝訊層’其具有可由—光碟閱讀機之—雷射讀取之一結 構；以及 至少一約微米級化學反應特徵，其係配置於該資訊層 中該光碟之一空間入射表面上，其中該至少一化學反應 特徵係與該可尋軌結構共焦配置。 12·如請求項11之光碟，其中該可讀取結構包括一擺動凹 槽。 13·如請求項11之光碟，其中該資訊層包括一反射層，且其 中該至少一化學反應特徵係配置於該反射層之一表面 上。 如請求項丨1之光碟，進一步包括·· 基板’邊基板具有一圖案化表面及一額外表面·， 以及 一反射層，其中該至少一化學反應特徵在該基板圖案 化表面在當該基板圖案化表面較該基板額外表面接近該 雷射時係配置為較該反射層接近該雷射。 15.如請求項14之光碟，其中該至少一化學反應特徵係配置 98815.doc -2- 1249158 於該反射層之一表面上。 16·如請求項1或丨丨之光碟，1 ^ ^ ^ ^ ^ " 至夕—化學反應特徵在 汶先碟之可哥軌讀取期間隹 17.如請求項16之光碟，1 / 了分辨信號。 號中一振幅變動。 係為同頻（HF)^ 18·如請求項16之光碟 (TE)信號中一振幅變 19 ·如請求項16之光碟 (FE)信號中一振幅變An operation structure, which is included in the (4) towel, the operation structure can be tracked by the laser when the patterned surface of the substrate is closer to a laser than the additional surface of the substrate; The chemical reaction characteristics of the operative structure can be read simultaneously. u 2·4] The optical disc of claim 1 wherein the operational structure is configurable to be a positive relief forward image. , , , , 3 · If the request item is terminated — the first disc, , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , A ^, the optical disc, wherein the operational structure is configurable to be 贞 予 予 前 forward image. 5. If the request item is 1 s first disc, the operating structure can be tracked: the surface closest to the extra surface of the substrate. 7. If the optical disc of the request is requested, the operational structure includes - a wobble track. The operation ~ 'the optical disc' wherein the plurality of chemical reaction characteristics are combined with the , , and " 8. If the layer is requested, and the φ, the optical disc, wherein the information layer further comprises - reflection, the plurality of chemical reaction features are in the substrate patterning 988I5.doc 1249158 额外 the additional surface of the substrate is close to the laser The time system is configured to be closer to the laser than the reflective layer. A monthly long disc of 1 wherein the information layer further comprises a reflective layer. The plurality of chemical reaction features of the member's disc are disposed on a spatial incident surface of the reflective layer. H--a searchable optical disc comprising: a beta layer having a structure that can be read by a laser-reading machine; and at least one micro-scale chemical reaction characteristic configured in the information One of the layers of the optical disk is incident on the surface of the optical disk, wherein the at least one chemically reactive feature is in a confocal configuration with the traceable structure. 12. The optical disc of claim 11, wherein the readable structure comprises a wobbled recess. 13. The optical disc of claim 11, wherein the information layer comprises a reflective layer, and wherein the at least one chemical reaction characteristic is disposed on a surface of the reflective layer. The optical disc of claim 1, further comprising: a substrate having a patterned surface and an additional surface, and a reflective layer, wherein the at least one chemically reactive feature is on the substrate patterned surface when the substrate pattern The surface of the laser is disposed closer to the laser than the additional surface of the substrate. 15. The optical disc of claim 14, wherein the at least one chemical reaction characteristic is configured to be 98815.doc -2- 1249158 on a surface of the reflective layer. 16·If Request Item 1 or 丨丨 丨丨 CD, 1 ^ ^ ^ ^ ^ " 夕夕—The chemical reaction characteristics are recorded during the reading of the Wenshou dish. 隹 17. As requested in the CD, 1 / Resolve the signal. The number one amplitude changes. Is the same frequency (HF) ^ 18 · The amplitude of the optical disc (TE) signal of claim 16 is changed 19 · The amplitude of the optical disc (FE) signal of claim 16 is changed ’其中該可分辨信號係為尋執錯誤 動。 '、中"亥可分辨信號係為聚焦錯誤 動0 20. —種多重資料層光碟，其包含·· 一反射層； 一額外反射層；以及 複數個化學反應特徵； *其中该反射層或該額外反射層具有可由一光學閱讀機 哥執之-表面，且其中—或多個該等化學反應特徵係配 置於該可尋軌結構中。 21·如請求項20之光碟，其中一或多個該等複數個化學反應 特U及该可尋執結構係可由該相同光碟閱讀機讀取的。 22·如請求項20之光碟，其中該額外反射層係為半反射的。 23·如請求項22之光碟，其中一或多個該等複數個化學反應 特欲係與該反射層或該半反射層之一表面共焦配置。 24.如凊求項23之光碟，其中一或多個該等複數個化學反應 特徵係與該半反射層之一表面共焦配置。 25· 一種光碟，其包括： ^88J5.doc 1249158 一基板，該基板具有一圖案化表面及_額外表面； 複數個化學反應特徵，其係與一操作結構共焦配置 以及 一資訊層； 其中該資訊層包括當該基板圖案化表面較該基板額外 表面接近-光碟之1射時配置於前向圖像正浮雕之該 操作結構。 • 26· 一光碟組件，其包括： 一光碟，其包括： 一資訊層； 一基板，其具有1案化表面及—額外表面，該圖案 化表面提供該資訊層； 一操作結構，其相關於該資訊I，該操作結構在當該 基板之該胃案化表面較縣板之該帛外表面接近一光碟 之一雷射時可由一光碟驅動器尋執的；以及 • 至少-化學反應特徵，其與該操作結構可同時讀取 的； -蓋板，其係可拆裝地加在該光碟上，該蓋板協助— 入射光束聚焦在該資訊層上。 988l5.doc -4-Where the distinguishable signal is a seek error. ',中"Hai-resolvable signal system is a focus error. 20. A multi-data layer optical disc containing a reflective layer; an additional reflective layer; and a plurality of chemical reaction features; The additional reflective layer has a surface that can be performed by an optical reader, and wherein - or a plurality of the chemical reaction features are disposed in the traceable structure. 21. The optical disc of claim 20, wherein one or more of said plurality of chemically reactive components and said configurable structure are readable by said same optical disc reader. 22. The optical disc of claim 20, wherein the additional reflective layer is semi-reflective. 23. The optical disc of claim 22, wherein one or more of said plurality of chemical reactions are in a confocal configuration with one of said reflective layer or one of said semi-reflective layers. 24. The optical disc of claim 23, wherein one or more of said plurality of chemical reaction features are confocal with a surface of one of said semi-reflective layers. 25. An optical disc comprising: ^88J5.doc 1249158 a substrate having a patterned surface and an additional surface; a plurality of chemical reaction features coupled to an operational structure in a confocal configuration and an information layer; The information layer includes the operational structure disposed in the forward image positive relief when the patterned surface of the substrate is closer to the additional surface of the substrate - the one of the optical discs. • A disc assembly comprising: a compact disc comprising: an information layer; a substrate having a patterned surface and an additional surface, the patterned surface providing the information layer; an operational structure associated with The information I is operable by a disc drive when the stomach surface of the substrate is closer to a laser of the disc than the outer surface of the panel; and • at least a chemical reaction characteristic, And the operating structure can be read simultaneously; a cover plate detachably attached to the optical disc, the cover assisting - the incident beam is focused on the information layer. 988l5.doc -4-