1300843 九、發明說明： 【發明所屬之技術領域】 本發明係有_-種生物晶片料方法及 其是一種可以增加生物晶片密度的點陣方法及其裝置 【先前技術】 在現代醫學的基礎研究或是臨床診斷中 =寅，Γ重的角色，尤其是隨著科技的進步 :==、分析的微陣列生物晶片，成為 一項新興的重要研究工具。 ~ 一般而言’微陣列生物晶片的製造方法，大體可區八 為原位合成法、接觸點墨法以及非接觸式时墨法，立戶: 需要的設備分別為半導體製程設備、微陣列晶片點陣機以 及=液設備，，些製程或設備均需要複雜的自動化控制 糸統’而且造價高昂。雖然有部分公司已開始 曰 入=其成本昂貴，產品售價仍居高不下，並不適： h驗至應用；加上中小型研究室均有其各自的研究計 晝’亚無廠商製造其所需的生物晶片，因此急需提供—種 方便操作的微陣列生物晶片製造方法及設備。 傳統的點陣機台，受限於某些機構上的限制，益法製 備較高密度的微陣列生物晶片，例如：試劑沾點針之間存 在有疋的間隔距離’所以沾點在載片上的試劑之間會有 同樣'間隔距離。雖然試劑沾點針的間距可以做的比較 小’但是由於試劑存放設備的限制，例如以96孔盤做為試 1300843 劑存放的設備，1 P屬曰A 4、2 m 因為W樣麵設襟，然而 心间心間距約為9公釐，若首接 由96孔盤作為沾點針的 1接 距必須設以9八,/ A 肢㈣劑沾點針的間 A 以免造成沾取試劑時的困擾。因此 =::以歸納出至少兩點無法製備較高密度生物 严曰門題.^為補沾點針本身因結構限制所產生的間 == 制即為沾點針間距必須要配合沾取試劑 是rf加微陣列晶片的試劑密度，有-種方法 2用早-的試劑沾點針’不斷地重複多次的户 ::題來;備微陣列晶片，雖然克服靖 、碭可以增加微陣列晶片的密度，但 ，須清洗試劑沾點針，以免造成污染而= =點-種試劑’因此製造速度緩慢，使用上十分的不方 另一種以人工方式操作的點陣機技術，主要如美國糞 號所示’係有關於一種提供多重= 間的點陣機。請參閱第-圖，點陣機⑽） 平板（11)、一第-早u… 弟一 沾cm )、一基底板（13)以及- j二⑴）所組成’基底板⑴）為容納載片的地方， :平板（11)、第二平板（12)以及基底板（1”上 句有’可供沾點器（18)穿過而將試劑沾點於载 ’沾點裔（18)則透過第二平板（12)表面的定位裝 1300843 置，以確保其下針沾點的位置。第一平板（11)可於基底 板（13 )上進行一第一方向〇d)的滑動，而第二平板（12) 則可於第-平板上（11)進行—第二方向（2d)的滑動， 其中在第一平板（11)與第二平板（12)表面，則分別具 有一系列的定位孔（17)。使用者可以透過插梢（16)插 入不同的定位孔（17)’使得沾點器（18)相對於載片表 面可以產生距之位移。#決定第—平板（ιι)與第 二平板（12)的位置後，使用者將沾點器（18)沾取試劑， 再把沾點器（18)定位於第二平板（12)上，而使試劑沾 點針（18a)得以在載絲面依序沾點試劑，以形成特定形 狀之微陣列。 運用刖述的裝置雖可增加微陣列晶片表面試劑的密 又使用者在知作時需要不斷地確認沾點器的移動方 =，以免造成試劑重疊而產生污染。此外，沾點器上的 劑沾點針仍受限於試劑存放設備之間距，Μ 9 為 孔之中心間距A Q /為 6孔盤之任兩個樣品 八敕且⑽‘、‘，^，如該裝置之沾點針針面直徑為0 5 行多達^次的沾點動作方能完成轉歹]日日片，則必須要進 職是之故’申請人鑑於習知技術 =心=研究，並，而不捨之精神，』= 發明之「生物晶片點陣 、構心出本 簡要說明。 去及/、衣置」，以下為本發明之 ⑧ 7 1300843 【發明内容】 本發明之構想係提供一種生物晶片點陣方法及其裝 置，尤其是一種可以增加生物晶片密度的點陣方法及其 置。 〆、/" 為了達成增加生物晶片表面試劑的點陣密度之目的， 本發明之-構想係提供—試劑盤，包含―試劑注入孔組、 4劑沾取孔組以及複數個微流道。該試劑盤之該複數個 微流道係用以分別連結各該試劑注入孔以及與其對應之各 該試劑沾取孔，以及該試劑沾取孔組巾各韻齡取狀 間距係小於該試劑注入孔組中各該試劑注入孔之間距。 依據本叙明之一貫施例，本發明之該試劑盤係由一上 層盤與一下層盤所組成。 較佳者，該上層盤表面具有該試劑注入孔與該試劑沾 取孔之開口。 較佳者，該下層盤表面對應於該上層盤之該試劑注入 孔人忒4劑沾取孔之該開口之處分別具有一凹槽，以容納 試劑。 較佳者，本發明之試劑盤之下層盤表面具有複數個微 流道以分騎結對應於上層盤之試劑注人孔與·沾取孔 之開口之凹槽。 較佳者，該上層盤表面係為疏水性材質或經過疏水性 處理。 較佳者，該下層盤表面係為親水性材質或經過親水性 處理。 1300843 —本發明之另一構想係提供一種生物晶片點陣裝置，包 -載片放置盤、一试劑盤以及一針座。該載片放置盤具 有至乂载片承座以接受—載片，以及該針座具有一試劑 沾點針組、1試劑盤係包含—試劑注人孔組、—試劑沾取 孔組以及複數個微流道；該試劑盤之該複數個微流道係用 以分別連^各該試劑注入孔以及與其對應之各該試劑沾取 孔上以及D亥试劑沾取孔組中各該試劑沾取孔之間距係小於 m主入孔組中各該試劑注入孔之間距。各該試劑沾點 沾取孔沾取試劑後，該針座移動至該議 劑沾點於該載絲面。 直彳將以 十=據本發明之一實施例’本發明之該載片承座具有— 疋位溝槽以容納該载片。 /、 動裝ΐ仏者’錢位溝槽包括一載片定位裝置與—載片滑 依::發明之另一實施例’該載片係先置於一定位載 -上，再置入該载片承座。 度，承座㈣^魅^具有微調刻 “彳載具可在該載片承座上滑動固定距離。 發明之一構想係提供另一 ”座、-載_盤以及一針座物 :广亥底座上’其具有一第一载片承 片糸 —第，以及-第二載片。= -n點針組以及一第二試劑沾點針組。該第 Ί300843 '及第_ 4劑沾點針誕係移動 分別對應於該第-載片承紅—帛=放置盤上方’以 第二载片承座之—第… —4劑沾點位置以及該 點於兮當 —式劑沾點位置，分別將各該試劑沾 =㈣之表面’該载片放置盤在各該試 I占點針組每次沾點後並轉動—預定角度，使各 座移動至次一試劑沾點位置。 σ 7、 =者，該第―_㈣針組與該第二試㈣ 係沾點在該載片表面之不重疊之位置。 τ、1300843 IX. Description of the Invention: [Technical Field] The present invention relates to a biochip material method and a lattice method and apparatus thereof capable of increasing the density of a biochip [Prior Art] Basic research in modern medicine Or in clinical diagnosis = 寅, a serious role, especially with the advancement of technology: ==, analysis of microarray biochips, has become an emerging important research tool. ~ Generally speaking, 'the manufacturing method of microarray bio-wafer, generally can be in-situ synthesis method, contact point ink method and non-contact time-of-flight method, stand: the required equipment is semiconductor process equipment, microarray wafer point Arrays and = liquid equipment, some processes or equipment require complex automation control systems and are expensive. Although some companies have begun to break in = their cost is high, the price of the product is still high, and it is not suitable: h to the application; plus small and medium-sized laboratories have their own research plan 'Asian no manufacturer to manufacture its The biochips required, there is an urgent need to provide a convenient microarray biochip fabrication method and apparatus. Conventional dot matrix machines, limited by certain institutional limitations, can be used to prepare higher density microarray biochips. For example, there is a sturdy separation distance between reagents and needles, so the spots are on the slides. There will be the same 'space separation distance' between the reagents. Although the distance between the reagent and the needle can be made relatively small, but due to the limitations of the reagent storage device, for example, a 96-well plate is used as the device for the storage of 1300,843, 1 P belongs to A 4, 2 m because the W sample is set. However, the distance between the centers of the heart is about 9 mm. If the first one is connected by a 96-well plate as a dip needle, the distance between the needles must be set to 9 or 8, and the A-A (four) agent should be placed between the needles to avoid the time when the reagents are taken. Troubled. Therefore, =:: can not be prepared to at least two points can not prepare a higher density of biological rigorous problems. ^ is the difference between the needle and the needle itself due to structural constraints == system is the needle spacing must be matched with the reagent Is the reagent density of the rf plus microarray wafer, there are - method 2 with the early - reagent dipping needle 'continuously repeat the number of households:: to prepare; micro-array wafers, although overcoming Jing, 砀 can increase the microarray wafer Density, however, must be cleaned with reagents and needles to avoid contamination and = = point - kind of reagents - so the manufacturing speed is slow, the use of very different methods, another manual operation of the dot matrix machine technology, mainly such as the US feces The number shown is related to a dot matrix machine that provides multiple =. Please refer to the figure - dot matrix machine (10). The flat plate (11), one first-early u... the first one (cm), one base plate (13) and - j two (1) are composed of 'base plate (1)) for receiving The place of the film: the flat plate (11), the second flat plate (12), and the base plate (1) have the phrase 'to allow the dipping device (18) to pass through and the reagent is dipped in the 'dense point (18) Then, the positioning device 1300843 is disposed on the surface of the second plate (12) to ensure the position of the lower needle. The first plate (11) can slide in the first direction (d) on the base plate (13). The second plate (12) can be slid in the second direction (2d) on the first plate (11), wherein the first plate (11) and the second plate (12) have a series of surfaces respectively. Positioning hole (17). The user can insert a different positioning hole (17) through the tip (16) to make the displacement of the spotter (18) relative to the surface of the slide. #定第第平板( ( After the position of the second plate (12), the user picks up the reagent from the spotting device (18), and then positions the spotting device (18) on the second plate (12) to make the reagent spotted. The needle (18a) can sequentially dipped the reagent on the surface of the carrier to form a microarray of a specific shape. The device used in the description can increase the density of the reagent on the surface of the microarray wafer, and the user needs to constantly confirm the stain when knowing it. The moving side of the pointer = to avoid contamination caused by overlapping reagents. In addition, the needle on the sticking device is still limited by the distance between the reagent storage devices, Μ 9 is the center spacing of the holes AQ / 6 holes If any two samples are gossip and (10)', ', ^, if the diameter of the needle surface of the device is 0 5 lines up to ^ times, the spotting action can complete the transfer] Japanese film, you must enter The job is the reason 'applicant in view of the prior art = heart = research, and the spirit of perseverance, 』 = "biochip dot matrix, constructive brief description. Go and /, clothing", below SUMMARY OF THE INVENTION The present invention is directed to a biochip dot matrix method and apparatus therefor, and more particularly to a dot matrix method and apparatus for increasing the density of a biochip.为了, /" In order to achieve the purpose of increasing the dot matrix density of the biochip surface reagent, the present invention is conceived to provide a reagent disk comprising a "reagent injection hole group, a 4-agent extraction hole group, and a plurality of microchannels. The plurality of microchannels of the reagent disk are respectively connected to the reagent injection holes and the corresponding reagent extraction holes corresponding thereto, and the reagent extraction hole groups are smaller than the reagent injection. The distance between each of the reagent injection holes in the well group. According to the consistent embodiment of the present invention, the reagent tray of the present invention consists of an upper tray and a lower tray. Preferably, the surface of the upper disc has an opening for the reagent injection hole and the reagent sinking hole. Preferably, the lower disc surface has a recess corresponding to the opening of the reagent injecting hole of the upper disc, respectively, to accommodate the reagent. Preferably, the surface of the tray of the reagent tray of the present invention has a plurality of microchannels for riding the grooves corresponding to the opening of the reagent injection hole and the extraction hole of the upper tray. Preferably, the surface of the upper disc is made of a hydrophobic material or subjected to a hydrophobic treatment. Preferably, the underlying disc surface is made of a hydrophilic material or subjected to a hydrophilic treatment. 1300843 - Another idea of the present invention is to provide a biochip dot matrix device comprising a carrier placement disk, a reagent disk and a hub. The slide placement tray has a carrier holder for accepting a carrier, and the needle holder has a reagent spot group, a reagent tray containing a reagent injection hole group, a reagent extraction hole group, and a plurality a plurality of microchannels; the plurality of microchannels of the reagent disk are respectively connected to each of the reagent injection holes and the corresponding reagent extraction holes and the Dhai reagent extraction hole group The distance between the holes of the pick-up holes is less than the distance between the injection holes of the reagents in the main-inlet group. After the reagent is spotted, the needle is moved to the surface of the carrier. The cymbal will be in accordance with one embodiment of the present invention. The carrier holder of the present invention has a clamping groove to accommodate the carrier. /, the mobile device's 'bit slot includes a slide positioning device and - slide slip: another embodiment of the invention 'the slide is placed on a positioning carrier - and then placed in the Slide carrier. Degree, bearing (four) ^ charm ^ with fine-tuning engraved "彳 carrier can slide a fixed distance on the carrier. One of the inventions is to provide another "seat, - load _ disk and a needle seat: Guanghai On the base, it has a first carrier sheet, a first, and a second carrier. = -n point needle group and a second reagent spot needle group. The movement of the third 843300843' and the _4 dose of the needles respectively correspond to the position of the first carrier-bearing red-帛=the top of the tray, the second carrier holder- The point is at the position of the jingle-type agent, respectively, and the surface of each of the reagents is smeared = (4). The slide is placed on each of the test I occupies the needle group after each spot and rotated - a predetermined angle, so that each Move the seat to the next reagent spot. σ 7, =, the first _ (four) needle group and the second test (four) system point on the surface of the slide does not overlap. τ,
==之另一實施例’該載片放置盤與該底座之 係故置有_定位裝置，使該載片放置射㈣ 轉動至固定位置。 X ϋ亥載片承座具有一定位溝 ϋ亥載片係置於一定位載具 依據本發明之另一實施例 槽以容納該載片。 依據本發明之另一實施例 上，再放入該載片承座。 本發明之另-構想係提供另一種生物晶片點陣装置， 包含-底座、-載 >；放置盤、—試缝以及—針座。該截 片，置盤係設置於該底座上，並具有_第—載片承座以及 一第二載片承座，以分別接受一第一載片以及一第二載 片。該試劑盤係包含一試劑注入孔組、一試劑沾取孔組以 及祓數個微流道；該試劑盤之該複數個微流道係用以分別 連結各該試劑注入孔以及與其對應之各該試劑沾取孔，以 及該試劑沾取孔組中各該試劑沾取孔之間距係小於該試劑 注入孔組中各該試劑注入孔之間距。該針座具有至少一試 1300843 劑沾點針組’其中各該試劑沾點針自各該試劑沾取孔沾取 试劑後，該針座移動至該載片放置盤上方對應於該第一载 f承座之♦試劑沾點位置，以將該試劑沾點於該第一 表^該载片放置盤在該試劑沾點針每次沾點後並 :力預疋ή度’使該第：制承絲動至 點位置以及該第—载片承座移動至該第二試劑沾點位^占 依據本發明之一實施例，該試劑盤包含-第-試劑注 入孔組與對應之-第—_沾取 入孔組與對應之—第二試劑沾取孔組，二 -输點針組以及一第二試劑沾點針組 與該第:試劑沾取孔組沾取試劑，並= 較二者1，该弟二試劑沾點位置沾點各該載片。 係沾點在該載U面之不重疊之位置髮點針組 間伟另—實施例’該載片放置盤與該底座之 鲁轉卿』可以預定角度 槽以另-實施例，該載片承座具有-定位溝 動裝t者，該定位溝槽包括-載片定位裝置與一载片滑 依據本發明之另一實施例於 上，再放人該载片承座。 载片係置於-定位载具 較佳者’該載片承座與該定位載具之間具有微調刻 11 1300843 又可在該載片承座上滑動固定距離。 的方共一種增力，晶片點陣密度 沾點位置，哕載片？、片於一载片承座上之-第-試劑 一試劑沾^ /設胁放錢上；將一第 劑=片t動至一第二沾點位置;以及將-第二試 士占點針組沾點至該載片表面。 ❿ 該載片放置錢以—預定角度轉動， 片表面之相鄰之不重心點物分別沾點在該載 依據本發明之一實施例，該載片係先置於一定位載具 上，再置入該載片承座上。 〃、 較，者°亥疋位載具與該載片承座之間具有微調刻 使D亥疋位载具可在该載片承座上滑動固定距離。 本發明之另一構想係提供另—種增加生物晶片點陣密 度的方法，包含：分別注人不同之試劑於—試劑盤之各試 劑注入孔中；安置一载片於一载片承座上；提供至少一試 劑沾點針組，以自各試咖取孔沾取試劑；移動該沾取試 劑之試劑沾點針組至該載片承座上方之—試劑沾點位置； 以及將该试劑沾點針組沾點至該載片表面。 依據本發明之一實施例，談試劑盤係包含一試劑注入 孔組、一試劑沾取孔組以及複數個微流道；該試劑盤之該 複數個微流道係用以分別連結各該試劑注入孔以及與其對 應之各该试劑沾取孔，以及該試劑沾取孔組中各該試劑沾 12 Ί300843 取孔之間距係小 距。 於該試劑注入孔組中各該試 劑注入孔之間 依據本發明之另一實施例， 片放μ上。 輕片承座係設置於一載 得棘Γ圭者’該載片放置盤在該試劑沾點針組每次沾點後 :::-預定角度’使該載片承座移動至次—沾點位置， 亚由:人-試劑沾點針組沾點於該載片表面。 =，各該試劑沾點針組係沾點在該载片表面之相 耶之不重登之位置。 依：本發明之另一實施例，該載片係先置於一定位載 /、上，再置入該載片承座上。 2佳者，該定位载具與該“承座之間具有微調刻 又使錢位载具可在該載片承座上滑動固定距離。 佳者’該定位載具在該試劑沾點針沾點載片後，於 j片备承座上滑動-預定距離，使該試劑沾點針在該載片 複進仃沾點，以製備—具有重複試劑之生物晶片。 =明曰中所稱之生物晶片，包括但不限於：微陣列晶 日日片、RNA晶片或蛋白質晶片。 本發明中所稱之試劑係指含有探針之溶液，該探針係 扣用於檢測生物樣本的生物 # '' ]生物活性分子，其種類包括但不限 NA、簡、寡核«、胜肽、蛋白質或其衍生物。 本發明中所稱之載片，係指將試劑沾點於其表面，而 用於製備生物晶片之基材，其材f包括但不限於：尼龍、 破璃、石英、％晶片、尼龍膜、以及各種高分子聚合物， 13 Ί300843 例如聚氯乙烯（polyvinylchloride，PVC )、聚乙烯 (polyethylene， PE )、聚曱基丙烯酸曱酯 (polymethylmethacrylate，PMMA )、聚胺基曱酸酯 (polyurethane， PU )、聚二曱基石夕氧烧 (polydimethylsiloxane，PDMS )、聚苯乙烯（polystyrene， PS )、聚碳酸酯（polycarbonate, PC )、聚四氟乙烯 (polytetrafluoroethylene, PTFE ) 〇 本發明中所稱之親、疏水性，係指試劑接觸材質表面 之相對性之親疏水現象。 本發明中所稱之親、疏水性處理，係指透過對材質表 面之處理，使該表面具有相對之親、疏水性，其方法包括 但不限於利用電漿處理與官能基接枝技術。 為了易於說明，本發明得藉由下述之較佳實施例及圖 示而得到充分暸解，並使得熟習本技藝之人士可以據以完 成之，然本發明之實施型態並不限制於下列實施例中。 【實施方式】 一般而言，當試劑沾點針的間距縮小時，雖然可增加 微陣列晶片表面的試劑密度，但卻不易充填試劑，而且容 易造成污染。因此本發明提供一容易充填試劑，又可有效 降低試劑沾點針間距之試劑盤。 請參閱第二圖，其為本發明之試劑盤之結構示意圖， 該試劑盤（20)係由一上層盤（21)與一下層盤（22)所 組成，兩者之間可採用熱壓接合技術、網印黏著技術、雙 14 1300843 面膠黏合技術等方式加以黏合。本發明之試劑盤之上層盤 與下層盤可為相同或不同之材質。可用於本發明之試劑盤 之材質包括但不限於：玻璃、石英、矽晶片、以及各種高 分子聚合物，例如聚氯乙烯（polyvinylchloride, PVC)、 聚乙浠（polyethylene，PE )、聚曱基丙烯酸曱酯 (polymethylmethacrylate，PMMA )、聚胺基曱酸酯 (polyurethane, PU )、聚二曱基石夕氧烧 (polydimethylsiloxane，PDMS)、聚苯乙稀（polystyrene, PS )、聚碳酸酯（polycarbonate，PC )、聚四氟乙稀 (polytetrafluoroethylene，PTFE)。較佳者，該下層盤之材 質為親水性之玻璃、石英或矽晶片，而該上層盤之材質則 為疏水性之苯乙烯（PS)、聚碳酸酯（PC)或聚四氣乙稀 (PTFE) 〇 5亥上層盤（21 )具有若干開口 ’分別為試劑注入孔（2 3 ) 以及試劑沾取孔（24)，而該下層盤（22)則具有若干凹 槽（25、26)以及微流道（27)。藉由微流道中的毛細現 象’使用者可自試劑注入孔（23 )充填試劑，試劑會流入 下層的凹槽（25)中，並經過微流道（27)流入凹槽（26) 中’而試劑沾點針則可自試劑沾取孔（24)伸入凹槽（26)， 並沾取試劑。本發明所提供之試劑盤中，試劑注入孔（23) 之間的距離較寬，有利於充填試劑，試劑沾取孔（24)之 間的距離較窄，則可以配合密度較高的針座，以製備較高 密度的微陣列晶片。 在微流道（27)中試劑流動的情形係決定於毛細現象、 15 ⑧ 1300843 微流道内壁面的平滑程度以及微流道表面的親疏水特性， =以電漿清洗處理下層盤（22)的表面，將其修都為親 水性，有助於試劑快速流動並可順暢流至所需位置。此外， 該上層盤（21)表面亦經過疏水性處理。 試f時，因為上、下層的親、疏水性不同的緣故 不曰沾黏在表面，而很容易流入下層盤（22)巾，並Another embodiment of the == mounting of the slide and the base is provided with a positioning means for rotating the slide (4) to a fixed position. The X ϋ 载 片 片 has a locating groove. The 载 载 片 is placed in a positioning carrier. According to another embodiment of the present invention, the carrier is accommodated. In accordance with another embodiment of the present invention, the carrier holder is placed. Another idea of the present invention is to provide another biochip dot matrix device comprising - a base, a - load >; a placement disk, a trial seam and a needle holder. The cutting piece is disposed on the base and has a first carrier holder and a second carrier holder for receiving a first carrier and a second carrier, respectively. The reagent tray comprises a reagent injection hole group, a reagent extraction hole group and a plurality of micro flow channels; the plurality of micro flow channels of the reagent disk are respectively connected to the respective reagent injection holes and corresponding ones thereof The reagent picking hole, and the distance between the reagent picking holes in the reagent picking hole group is smaller than the distance between the reagent injection holes in the reagent injection hole group. The needle holder has at least one test 1300843 dose spotting needle set, wherein each of the reagent dipping needles is taken from each of the reagent picking holes, and the needle holder moves to the top of the slide placing tray corresponding to the first load f The position of the reagent ♦ is stained to the point where the reagent is spotted on the first table. The slide is placed on the tray after the reagent is spotted by the needle and the force is pre-twisted to make the first: The wire-moving-to-point position and the movement of the first carrier-carrying seat to the second reagent-spotting position. According to an embodiment of the present invention, the reagent disk comprises a --reagent injection hole group and a corresponding - - _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ In the first two, the two reagents are spotted and spotted on each of the slides. The spotting point is between the positions of the non-overlapping positions of the U-faces, and the other embodiments of the embodiment, the carrier placement disk and the base of the base can be predetermined angle grooves to another embodiment, the slide The socket has a positioning groove, and the positioning groove includes a carrier positioning device and a carrier slide according to another embodiment of the present invention, and the carrier holder is placed. The carrier is placed on the positioning carrier. Preferably, the carrier holder and the positioning carrier have a fine adjustment 11 1300 843 and a fixed distance on the carrier holder. The total side of the force, the density of the wafer dot-dot position, 哕 slide? , the film on a carrier holder - the first reagent - reagent dip ^ / set the risk on the money; a first dose = piece t to a second spot position; and will - second tester account The needle set is spotted onto the surface of the slide. ❿ the slide is placed at a predetermined angle, and the adjacent non-centered points of the surface of the sheet are respectively dipped in the carrier. According to an embodiment of the present invention, the slide is first placed on a positioning carrier, and then Placed on the carrier holder. 〃 较 较 ° ° ° ° ° ° 疋 疋 疋 疋 疋 疋 疋 疋 具有 具有 具有 具有 具有 具有 具有 具有 具有 具有 具有 具有 具有 具有 具有 具有 具有 具有 具有 具有 具有 具有 具有 具有 具有 具有Another idea of the present invention is to provide another method for increasing the density of a biochip dot matrix, comprising: separately injecting different reagents into each reagent injection hole of the reagent disk; and placing a carrier on a carrier holder. Providing at least one reagent spotting needle set to take the reagent from each test coffee taking hole; moving the reagent of the picking agent to the spotting needle set to the reagent spot position above the carrier holder; and the reagent Dip the needle set to the surface of the slide. According to an embodiment of the invention, a reagent tray comprises a reagent injection hole group, a reagent extraction hole group and a plurality of microchannels; the plurality of microchannels of the reagent tray are used to respectively connect the reagents The injection hole and the corresponding reagent smear hole corresponding thereto, and the reagent immersing hole group each of the reagent immersed between the Ί300843 and the hole are separated by a small distance. Between each of the reagent injection holes in the reagent injection hole group, according to another embodiment of the present invention, the wafer is placed on the substrate. The light film bearing system is set in a carrier that is loaded with the carrier. The slide is placed on the needle after the reagent is applied to the needle group:::- predetermined angle to move the carrier to the secondary Point position, sub-man: The human-reagent spotted needle group is spotted on the surface of the slide. =, each of the reagents is spotted on the surface of the slide. According to another embodiment of the present invention, the slide is first placed on a positioning carrier, and placed on the carrier holder. 2 good, the positioning carrier and the "station between the seat has a fine-tuning engraving so that the money carrier can slide a fixed distance on the carrier. The good one of the positioning vehicle in the reagent dipped needle After the slide is loaded, the slide is placed on the j-seat holder for a predetermined distance, so that the reagent is spotted on the slide, and the biochip is prepared with a repeating reagent. Biochips, including but not limited to: microarray crystals, RNA wafers or protein wafers. The term "reagent" as used in the present invention refers to a solution containing a probe that is used to detect a biological sample. '] a biologically active molecule, the species including but not limited to NA, simple, oligonuclear «, peptide, protein or derivative thereof. The carrier referred to in the present invention means that the reagent is spotted on the surface thereof, and For preparing a substrate for a biochip, the material f includes, but is not limited to, nylon, glass, quartz, % wafer, nylon film, and various high molecular polymers, 13 Ί 300843, for example, polyvinyl chloride (PVC), polyethylene. (polyethylene, PE), polydecyl methacrylate (polymethylmethacrylate, PMMA), polyurethane (PU), polydimethylsiloxane (PDMS), polystyrene (PS), polycarbonate (PC), poly Polytetrafluoroethylene (PTFE) 亲 The affinity and hydrophobicity referred to in the present invention refers to the relative hydrophobicity of the relative contact of the reagent on the surface of the material. The affinity and hydrophobic treatment referred to in the present invention means that the pair is transparent. The surface of the material is treated to make the surface relatively hydrophilic and hydrophobic, and the method includes, but is not limited to, using a plasma treatment and a functional group grafting technique. For ease of description, the present invention is provided by the following preferred embodiments and The drawings are fully understood and can be accomplished by those skilled in the art, but the embodiments of the present invention are not limited to the following examples. [Embodiment] In general, when the reagent is spotted When the pitch is reduced, although the reagent density on the surface of the microarray wafer can be increased, it is not easy to fill the reagent, and it is easy to cause pollution. Therefore, the present invention provides a The reagent tray which is easy to fill the reagent and can effectively reduce the needle spacing of the reagent. Please refer to the second figure, which is a schematic structural diagram of the reagent tray of the present invention, the reagent tray (20) is composed of an upper tray (21) and a lower layer. The layer disc (22) is composed of two parts, which can be bonded by hot pressing bonding technology, screen printing adhesive technology, double 14 1300843 surface adhesive bonding technology, etc. The upper tray and the lower layer disc of the reagent tray of the present invention can be the same. Or different materials. The materials of the reagent trays that can be used in the present invention include, but are not limited to, glass, quartz, germanium wafers, and various high molecular polymers, such as polyvinyl chloride (PVC), polyethylene (PE), polyethylene (PE). ), polymethylmethacrylate (PMMA), polyurethane (PU), polydimethylsiloxane (PDMS), polystyrene (PS), poly Polycarbonate (PC), polytetrafluoroethylene (PTFE). Preferably, the material of the lower layer is a hydrophilic glass, quartz or germanium wafer, and the material of the upper layer is hydrophobic styrene (PS), polycarbonate (PC) or polytetraethylene ( PTFE) 〇5 上 upper tray (21) has a plurality of openings 'respectively for the reagent injection hole (23) and the reagent extraction hole (24), and the lower tray (22) has a plurality of grooves (25, 26) and Micro flow channel (27). By the capillary phenomenon in the microchannel, the user can fill the reagent from the reagent injection hole (23), and the reagent flows into the lower groove (25) and flows into the groove (26) through the micro flow channel (27). The reagent spotting needle can be inserted into the groove (26) from the reagent taking hole (24) and the reagent is taken up. In the reagent tray provided by the invention, the distance between the reagent injection holes (23) is wide, which is favorable for filling the reagent, and the distance between the reagent extraction holes (24) is narrow, and the needle holder with higher density can be matched. To prepare higher density microarray wafers. The flow of the reagent in the microchannel (27) is determined by the capillary phenomenon, the smoothness of the inner wall surface of the 15 8 1300843 microchannel, and the hydrophobicity of the surface of the microchannel, = the plasma treatment of the lower tray (22) The surface is repaired to be hydrophilic, helping the reagent to flow quickly and smoothly to the desired location. In addition, the surface of the upper tray (21) is also subjected to hydrophobic treatment. When testing f, because the upper and lower layers are different in affinity and hydrophobicity, they do not stick to the surface, but easily flow into the lower layer (22) towel, and
道(27)流入凹槽(26)備用。由於上:盤 )表面為疏水性，當試劑沾點針沾取試劑時，不 試劑殘留在試劑沾取孔（24)，而且，下層盤 : f親，特性可以保持試劑液面的高度。經由這樣：設 ^只要試劑沾點針每次下針沾取的深度—致，絲心 的試劑量是均等的，如此可以使每次沾點於载 片的6式劑1的恒定。 放置圖’其為依據本發明之較佳實施例之載片 晶片密度與欲沾點載二；之不=座照 載片—.μ j而承叹汁不冋數目的 片Γ ^貫施例的載片承座係以4個為例。為了將載 容％載Π，在載片承座（32)上’可採用—凹槽來 载片’或疋以固定座（33、34)來夾住載片。 ^閱第四圖，其為本發明之載片承座之另一實施 (42 :承座（4〇)係包括一底盤⑷）及载片固定座 固疋座（42、43)可從兩側夾住载片（未顯 ，/、内側並分別具有定位溝槽（44)以容納载片，甚 16 1300843The track (27) flows into the groove (26) for standby. Since the surface of the upper: disk is hydrophobic, when the reagent is applied to the reagent, the reagent remains in the reagent extraction hole (24), and the lower layer: f, the characteristic can maintain the height of the reagent liquid level. By this, it is assumed that the amount of the reagent of the silk core is equal as long as the depth at which the reagent is applied to the needle, and the amount of the reagent of the silk core is equal, so that the dose of the type 6 of the tablet can be made constant every time. The placement diagram 'is the density of the wafer of the wafer according to the preferred embodiment of the present invention, and the number of the wafers to be smudged. The carrier holders are based on four examples. In order to load the carrier %, a carrier can be used on the carrier holder (32) to hold the carrier or the holder (33, 34) to clamp the carrier. 4, which is another embodiment of the carrier holder of the present invention (42: the bearing (4〇) includes a chassis (4)) and the carrier fixing seat (42, 43) can be from two The side clamps the slide (not visible, /, inside and has a positioning groove (44) to accommodate the slide, even 16 1300843
三次丄從而可以製備三重複的微陣列晶片。 至可在溝槽内部設置定位裝置與滑動裝 辅助載片的敘叙.e k i ^ 請參閱第五@，其為本發明之針紅示意圖，從外觀 ^可見该針座（5〇)表面具有複數個試劑沾點針⑼的 ，放孔（51)，各擺放孔的位置需㈣事先預定的晶片密 度，與本發明之試劑盤搭配設置。 圖，其係為本發明之生物晶片點陣機之實 遺生物晶片點陣機（6〇 )包括底座（6工）， 請參閱第六圖 施例之示意圖，該 在，上設置有載片放置盤（62)以及針座（⑷。針座（⑷ 則，又置在-操作把手（65)上，並透過設置於底座⑷） 之間移動，該操作把手（65)可用人工方式在滑執上推動， 也可採用步進馬達為自動式操作，亦可使S —機械手臂來 操控針座（64)的移動。載片（63)則放置於載片放置盤 (62)上之载片承座中。 將已經沾取試劑後的針座（64)移動至載片放置盤（62) 的上方，將載片承座轉到對應的沾點位置，將針座（64 ) 向下壓以在載片（63)表面進行沾點的動作；將針座（64) 放開、歸位，並沿著旋轉方向（66)轉動載片放置盤（62)， 使下一個載片承座轉到沾點位置。重覆進行前述之沾取、 1300843 沾點動作，直到完成生物晶片的製備。 盤以片其=本發明之試劑盤，放置 晶片點陣機(7。)：二:=實施例之示意圖，該 般（74)二具有一底座（71)，以在其上設置試劑 ^ 载片放置盤（72)以及針座（75)，其中 (72) ^ 坐5)則設置在—操作把手（76)上，並透過設 於底座（71)兩侧邊的滑動裝置（例 ： =兩端點之間移動，該操作把手㈤= 2二=也可採用步進馬達為自動式操作，亦可 使用^械手臂來操控針座（75)的移動。 =整=點陣過程可由充填試劑盤開始。將試劑盤⑺） 7 ，坐71)上，並以—般實驗室常用的微量吸量管 (P.ette)^ θ因為相㈣親水性㈣道的設計，快速 取孔内。除了—般形式的微量吸二 ^由於提供網雜賴社 較大，可依需求提供不同排列的試 = 通用的4流道或8泣、音AAa^ L以配口目刖 以最便利的方式進:===使用’讓使用者能 充填試劑後，可透^Γ; 操作的方便性。 上的針座⑺）移動到試劑盤（74)的上\ =把手（76) 毛細作用，試劑沾點斜人Ώ ^此％因為 針冒吸入-定量的試劑，也由於試劑 ⑧ 18 1300843 盤$(74)表面親疏水性變化的設計，使得在試劑沾取孔内 4劑液面的高度會保持恒定，因此各試劑沾點針每次吸取 的试劑量也會維持一定量。Three replicates of the microarray wafer can be prepared three times. To the inside of the groove, the positioning device and the slide-mounted auxiliary slide are described. eki ^ Please refer to the fifth @, which is a schematic diagram of the needle red of the present invention, and the surface of the needle seat (5〇) has a plurality of The reagents are spotted on the needles (9), and the holes (51) are placed. The positions of the respective holes are required to be (4) predetermined wafer densities, and are arranged in combination with the reagent tray of the present invention. The figure shows that the biochip dot matrix machine (6〇) of the biochip dot matrix machine of the present invention comprises a base (6 working), please refer to the schematic diagram of the sixth embodiment, where the slide is arranged The disc (62) and the hub ((4). The hub ((4) is placed on the -operating handle (65) and moved between the base (4)), the operating handle (65) can be manually slipped The push motor can also be used for automatic operation, and the S-mechanical arm can be used to manipulate the movement of the needle holder (64). The slide (63) is placed on the slide placement plate (62). In the holder. Move the needle holder (64) after the reagent has been taken to the top of the slide placement tray (62), turn the carrier holder to the corresponding spot position, and lower the needle holder (64). Pressing the spot on the surface of the slide (63); releasing the needle seat (64), homing, and rotating the slide placement plate (62) in the direction of rotation (66), so that the next slide bearing The seat is turned to the spot position. Repeat the above extraction, 1300843 touch point action until the preparation of the biochip is completed. The reagent disk of the Ming, the wafer dot matrix machine (7.): two: = schematic diagram of the embodiment, the general (74) two has a base (71) to set the reagent ^ slide placement disk (72) thereon and The needle holder (75), wherein (72) ^ sitting 5) is disposed on the operating handle (76) and through the sliding device provided on both sides of the base (71) (eg: = moving between the two ends, The operation handle (5) = 2 2 = can also be automatically operated by stepper motor, or the arm can be used to control the movement of the needle seat (75). = The whole process of the dot matrix can be started by filling the reagent disk. (7)) 7 , sit on 71), and use the micro-pipette (P.ette) ^ θ commonly used in the laboratory to quickly take the hole because of the phase (four) hydrophilic (four) channel design. In addition to the general form of micro-absorption II, due to the large size of the network, we can provide different arrangements according to the needs of the test = general 4 flow channels or 8 weeping, sound AAa ^ L to match the mouth in the most convenient way In: === Use 'to allow users to fill the reagents, can be transparent; easy to operate. The needle seat (7) on the upper part moves to the upper side of the reagent plate (74). \=Handle (76) Capillary action, the reagent is dimmed. This is because the needle is a suction-quantitative reagent, and also because of the reagent 8 18 1300843 (74) The design of the surface hydrophobicity change is such that the height of the liquid level of the four doses in the reagent picking hole is kept constant, so that the amount of the reagent taken by each reagent sticking needle is maintained at a certain amount.
接下來將沾取試劑後的針座（75)移動至載片放置盤 (72)的上方’將載片承座轉到對應的沾點位置，將針座 (75 )向下壓以進行沾點的動作；將針座（75)放開、歸 並沿著旋轉方向⑺）轉動載片放置|(72)，使下 一個載片承座轉到沾點位置。重覆進行前述之沾取、沾點 動作’直到完成生物晶片的製備。 在本實施例中，針座（75)係固定在操作把手（76) 上，、透過一個滑動裝置（例如滑軌），在試劑盤（74)以 及载片放置盤（72)之間往覆移動（沿著方向77)。 透過本發明之增加生物晶片密度的點陣方法，製備微 片之流程請參閱第八圖之示意圖。以製備—個16點 、、政陣列晶片為例，所準備的試劑盤、針座各自有4個區 ^每個區域分別有4個試劑沾取孔或位置對應之4個試 =點針。將載片轉動到第—沾點位置，此時假設第一組 二剎沾點針在載片表面沾點的位置為a，再將载片放置盤 疋轉使該載片移動到第二沾點位置，此時1 = 4+ A却u 士 丁币—組戎劑沾點 、’卞在載片表面沾點的位置為在a點旁邊的b，二者不重聂· 重覆此-沾點、旋轉的過程，最後會形成如第人 物晶片。由第七圖的例子可以看到，各組試劑二 …十斤沾點的位置是互相交錯而不重複的，而且交 置則是事先依照所需求的密度，在針座上排列其位^。例 ⑧ 19 1300843 ^要形成前面所述的16點微陣列晶片，可以在針座 为成4個區域’每區4點的方式進行沾點丨若 抑 2陣列晶片’可以在針座上劃分4個區域，每區Μ點的 ^行沾點，依此類推，可以依照不同的密度需求安排 ^的針座以其與其搭配之試劑盤，其劃分的區域跟每區 或沾點的點數都可以依照實際的需求而改變。 本!5明之生物晶片點陣裝置，同時提供兩種方式以辦 ·=晶=面之點陣密度’―為利用本發明所提供之試^ m為利用本發明所揭露之點陣方法 沾點針於排列設計時，能增加試劑沾點針之間的密；允^ =留充填試麟之制性，·後者則是利用料_點、 ，之以’使得試劑沾點針組交錯地在載片表面進行沾 點’以快速增加生物晶片表面之沾點密度。 需要注意的是，本發明所提供之試劑盤、針座以及載 片放置盤可以分別供不同的點陣裝置使用，也可以植人而 ’成為本發明之生物晶片點陣機。例如，； 性之試劑盤，可依照不同的密度«，提 = 縣填試.用，而其_較窄的試劑 ==Γ其似劑沾點針之間具有相同間距的 成為成Α太、纟明之4劑盤、針座以及載片放置盤搭配 成為成為本發明之生物晶片點陣裝置時，可依，昭使 需求，亦即用於製造不同密度的生物晶片，而事先安排盆 式劑注入孔組、試劑沾取孔組以及針座的密度、載片承座 20 1300843 的密度及其旋轉角度，以製備所需密度的生物晶片。 本發明得由熟悉本技藝之人士任施匠思而為諸般修 飾，然皆不脫如附申請專利範圍所欲保護者。 Ί300843 【圖式簡單說明】 第一圖為習知之手動式點陣機之示意圖。 第二圖為依據本發明之實施例之試劑盤之結構示意 圖。 第三圖為依據本發明之實施例之載片放置盤及載片承 座之示意圖。 第四圖為依據本發明之實施例之載片承座與載片載具 之示意圖。 弟五圖為依據本發明之貫施例之針座之不意圖。 第六圖為依據本發明之一實施例之生物晶片點陣機之 示意圖。 第七圖為依據本發明之另一實施例之生物晶片點陣機 之示意圖。 第八圖為依據本發明之方法製備微陣列晶片之流程示 意圖。 1300843 【主要元件符號說明】 10點陣機 12第二平板 16插梢 18沾點器 Id第一方向 20試劑盤 22下層盤 24試劑沾取孔 27微流道 31底盤 33、34固定座 41底盤 44定位溝槽 46凹槽 50針座 52試劑沾點針 61底座 63載片 65操作把手 67移動方向 71底座 73旋轉方向 75針座 77移動方向 11第一平板， 13基底板 17定位孔 18a試劑沾點針 2d第二方向 21上層盤 23試劑注入孔 25、26凹槽 30載片放置盤 32載片承座 40載片承座 42、43固定座 45載片載具 47定位刻度 51擺放孔 60生物晶片點陣機 62載片放置盤 64針座 66旋轉方向 70生物晶片點陣機 72載片放置盤 74試劑盤 76操作把手 23Next, move the needle holder (75) after the reagent is taken to the top of the slide placement tray (72). Turn the carrier holder to the corresponding spot position and press the needle holder (75) down to dip. Point action; release the needle holder (75) and turn it in the direction of rotation (7) to rotate the slide placement|(72) to move the next slide holder to the spot position. The aforementioned picking and dipping operations are repeated until the preparation of the biochip is completed. In this embodiment, the needle hub (75) is fixed to the operating handle (76) and is passed between the reagent disk (74) and the carrier placement disk (72) through a sliding device (such as a slide rail). Move (along direction 77). For the flow of preparing the microchip by the dot matrix method of increasing the density of the biochip of the present invention, please refer to the schematic diagram of the eighth drawing. Taking a preparation of a 16-point, political array wafer as an example, the prepared reagent disk and the needle holder each have 4 zones. ^ Each zone has 4 reagent extraction holes or 4 test points corresponding to the position. Rotate the slide to the position of the first spot. At this time, assume that the position of the first set of two brakes on the surface of the slide is a, and then place the slide on the slide to move the slide to the second touch. Point position, at this time 1 = 4 + A but u 士丁币 - group 戎 沾 、 , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , The process of rotation will eventually form a wafer of the first character. As can be seen from the example of the seventh figure, the positions of the two groups of reagents are staggered and not repeated, and the intersection is arranged in advance on the needle holder according to the required density. Example 8 19 1300843 ^To form the 16-point microarray wafer described above, it is possible to divide the needle holder into 4 areas '4 points per area. 2 array wafers' can be divided on the needle holder 4 Areas, points in each area, and so on, can be arranged according to different density requirements. The needle holder is matched with the reagent tray, and the divided area and the number of points in each area or touch point are Can be changed according to actual needs. The biochip dot matrix device of the present invention provides two ways to do the dot matrix density of the surface of the crystal. The test is provided by the dot matrix method disclosed by the present invention. When the needle is arranged in the design, it can increase the density between the needles of the reagents; allow the filling of the test lining, and the latter use the material _ point, so that the reagents are scattered in the needle group. The surface of the slide is spotted to quickly increase the density of the spot on the surface of the biochip. It should be noted that the reagent disk, the needle holder and the carrier placement disk provided by the present invention can be used for different dot matrix devices, respectively, and can also be implanted into the biochip dot matrix machine of the present invention. For example, the reagent tray can be tested according to different densities, and the reagents with narrower reagents == the same spacing between the needles and the needles. When the four-disc tray, the needle holder and the slide placement disc of the invention are combined to form the bio-wafer dot matrix device of the present invention, it can be used to manufacture biochips of different densities, and the potting agent is arranged in advance. The injection well set, the reagent take-up set and the density of the hub, the density of the slide holder 20 1300843 and its angle of rotation are used to prepare a biochip of the desired density. The present invention has been modified by those skilled in the art and is intended to be modified as described in the appended claims. Ί300843 [Simple description of the diagram] The first picture is a schematic diagram of a conventional manual dot matrix machine. The second drawing is a schematic view showing the structure of a reagent disk according to an embodiment of the present invention. The third figure is a schematic view of a slide placement tray and a carrier holder in accordance with an embodiment of the present invention. Figure 4 is a schematic illustration of a slide carrier and a carrier carrier in accordance with an embodiment of the present invention. The fifth figure is a schematic view of the needle holder according to the embodiment of the present invention. Figure 6 is a schematic illustration of a biochip dot matrix machine in accordance with an embodiment of the present invention. Figure 7 is a schematic illustration of a biochip dot matrix machine in accordance with another embodiment of the present invention. The eighth figure is a schematic illustration of the process for preparing a microarray wafer in accordance with the method of the present invention. 1300843 [Main component symbol description] 10 dot matrix machine 12 second plate 16 pin tip 18 dip device Id first direction 20 reagent plate 22 lower plate 24 reagent dip hole 27 micro flow channel 31 chassis 33, 34 fixed seat 41 chassis 44 positioning groove 46 groove 50 needle seat 52 reagent spot needle 61 base 63 slide 65 operation handle 67 moving direction 71 base 73 rotation direction 75 needle seat 77 moving direction 11 first plate, 13 base plate 17 positioning hole 18a reagent Dip needle 2d second direction 21 upper plate 23 reagent injection hole 25, 26 groove 30 slide placement disk 32 carrier holder 40 carrier holder 42, 43 fixing seat 45 carrier carrier 47 positioning scale 51 placed Hole 60 biochip dot matrix machine 62 slide placement disk 64 needle holder 66 rotation direction 70 biochip dot matrix machine 72 slide placement disk 74 reagent plate 76 operation handle 23