TWM290505U - Automatic device used in nucleus manipulation - Google Patents
Automatic device used in nucleus manipulation Download PDFInfo
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- TWM290505U TWM290505U TW94220790U TW94220790U TWM290505U TW M290505 U TWM290505 U TW M290505U TW 94220790 U TW94220790 U TW 94220790U TW 94220790 U TW94220790 U TW 94220790U TW M290505 U TWM290505 U TW M290505U
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M290505 、創作說明(1) 新型7斤屬術領域】 一~^^ 本創作係有關一種「細胞核操作的自動化# 指一種將哺乳類動物之雌性生殖細胞(卵子 f置」,尤 細胞,以人工的方式進行體外之操作行為,、與成熟的體 胞相同D N A之無性繁殖者。、 、取待與體細 【先前技術】 按’傳統習知的細胞核操作均以倒置a§他& 二作機械臂搭載波璃固定針、玻璃注射針進彳-=,左右側 絲=進行卵細胞與體細胞的搜尋、固定、:動機械操 Ϊ中U、置入、注射、抽離、融合/激^的H移動、旋 雌性生瘦細胞之透明帶較厚,因此需的再循^動作, ☆光學式光刀在透明帶上行穿:以配化學酵 限於倒置龍進入雌性生殖細胞中;惟,前述之明A以使體 像,無法即,鏡之光學機構問題,只能取得X、γ平=去文 射時間的限fl進行Z軸量測,且受限於細胞核與UV螢Λ 目標,故鮮 ’、使細胞核操作一直無法達到「自動先Μ 創作人^ 之問題有再研發解決之道的必要性化」的 化」操作之鑑於上述習知細胞核的操作無法達到「 年的經驗,故,,爰精心研究,並積個人從事該項事案, 置,以解決、7 1作出一種勒新的細胞核操作的自動化^夕 本創作自動化」盲點,增加產能,提昇效率。、 置,其係利^主要目的在提供一種細胞核操作的自動化 與培養液後放二個具有微流體的玻片載入卵細胞、體細月^ 硬置於一自動化操作運動單元設備上,再利用M290505, creation instructions (1) New 7 jins field] 1~^^ This creation is related to a kind of "automation of nuclear operation" refers to a kind of female germ cells (egg f) of mammals, especially cells, artificial Ways to perform in vitro manipulations, asexual breeders of the same DNA as mature somatic cells, and to be fine-grained [pre-technical] According to the 'traditional conventional nuclear operation, all are inverted a§ he& The robot arm is equipped with a glass fixed needle, a glass injection needle into the 彳-=, and the left and right side wires are used for searching and fixing egg cells and somatic cells, and U: mechanical placement, injection, extraction, fusion/excitation The movement of H, the translucent female thin cells have thicker transparent bands, so it is necessary to follow the action, ☆ optical light knife is worn in the transparent belt: the chemical fermentation is limited to the inverted dragon into the female germ cells; Ming A, in order to make the body image, can not be the optical mechanism of the mirror, can only obtain the X-axis measurement of the X, γ flat = de-text time limit, and is limited by the nuclear and UV fluorescein target, so fresh ', the nuclear operation has been no In view of the fact that the problem of "automatic screed creator ^ has the need to re-develop and solve the problem", the operation of the above-mentioned conventional nucleus cannot achieve the "year of experience, so, carefully research, and personally Engaged in the case, set, to solve, 7 1 to create a new nuclear operation of the automation of the eve of the creation of the blind spot, increase productivity, improve efficiency., set, its main purpose is to provide a nuclear operation After automating and culturing, two slides with microfluids are loaded into the egg cells, and the body is finely placed on an automated operation unit.
M290505 四、創作說明(2) 一組雙軸顯微鏡擷取XY與XZ的平面影像反定出印細|¥在1 軸的高程位置,再利用設置於垂直顯微鏡正下方之雷射光 機電單元進行對卵細胞穿孔、去核,以及牽引與注射體細 胞之操作,最後可以光學融合或以化學激活,而達到自動 化操作細胞核之功效者。 【新型内容】 為達上述之目的,本創作「細胞核操作的自動化裝 置」主要構造包括由一自動化操作運動單元、一三向自由 度運動單元、一雙軸顯微鏡單元、一雷射光機電單元及一 電腦控制單元所組成;其特徵在:該自動化操作運動單元 0係在一機架上設有一橫移機構及一具有載玻台之縱移機 構;三向自由度運動單元包含一吸盤式機械臂與一橫向線 ' 性滑執,而可將在待料培育箱内之玻片移至自動化操作運 . 動單元的栽玻台,再由載玻台將完成操作後之玻片移至成 品培育箱,且其中之玻片的上表面具有一橫向的微流體, ^ 該微流體之寬度及深度略大於卵細胞之直徑;雙軸顯微鏡 - 單元係固定在自動操作運動單元之橫移機構上,其包括一 由上而下的垂直視頻顯微鏡、一由後向前的水平視頻顯微 鏡及一側向螢光燈組成,該二顯微鏡成9 0度排列且鏡頭對 向載玻台,且每一顯微鏡均包含有一 CCD攝影機及一同軸 射出雷射二極體;雷射光機電單元其至少包含有一雷射二 極體、一脈衝雷射機及一射出聚焦鏡,該雷射二極體與脈 衝雷射機係固定於自動化操作運動單元之機架底部,而可 經同一路徑傳送至射出聚焦鏡後向外照射;該射出聚焦鏡 (§)M290505 IV. Creation Instructions (2) A set of two-axis microscopes captures the XY and XZ plane images to determine the prints. ¥ In the elevation position of the 1 axis, use the laser electromechanical unit located directly below the vertical microscope. Oocyte perforation, enucleation, and manipulation of injecting and injecting somatic cells can be achieved by optical fusion or chemical activation to achieve automated function of the nucleus. [New Content] In order to achieve the above objectives, the main construction of the "automated device for nuclear operation" includes an automated operation unit, a three-directional degree of freedom motion unit, a dual-axis microscope unit, a laser electromechanical unit and a The computer control unit is composed of: the automatic operation motion unit 0 is provided with a traverse mechanism and a vertical movement mechanism with a glass table on a frame; the three-direction freedom movement unit comprises a suction cup type mechanical arm With a horizontal line, the slides in the waiting box can be moved to the glass table of the automated operation unit, and then the slides after the operation are moved to the finished product cultivation by the glass table. a box, and wherein the upper surface of the slide has a lateral microfluid, ^ the width and depth of the microfluid are slightly larger than the diameter of the egg cell; the biaxial microscope - the unit is fixed on the traverse mechanism of the automatically operated motion unit, It consists of a top-down vertical video microscope, a back-and-forward horizontal video microscope, and a side-by-side fluorescent lamp. The two microscopes are arranged at 90 degrees and the lens is arranged. a pair of glass slides, each of which includes a CCD camera and a coaxial injection laser diode; the laser electromechanical unit includes at least one laser diode, one pulse laser and one injection focusing mirror. The laser diode and the pulse laser are fixed to the bottom of the rack of the automated operation motion unit, and can be transmitted to the exit focusing mirror through the same path and then radiated outward; the exit focusing mirror (§)
第6頁 M290505 四、創作說明(3) 則係固定於自動化操作運動單元之横移機構上對應於垂直 視頻顯微鏡下方由下向上照射,且與垂直視頻顯徵鏡同 軸;電腦控制單元係用來與其他個單元訊號連線,並藉以 控制各單元自動運作者。 如上所述裝置使用時,係利用一個具有微流體的玻片 載入卵細胞、體細胞與培養液於其中之微流體之後,藉三 向自由度運動單元將數個玻片橫列的放置於一自動化操作 運動單元之載玻台上,再利用雙軸顯微鏡單元的垂直及水 平視頻顯微鏡擷取在微流體内卵細胞之XY與XZ的平面影 像,傳輸到電腦控制單元顯示其影像並計算出卵細胞在z 軸的高程位置,再利用設置於垂直顯微鏡正下方之雷射光 機電單元,配合電腦自動調整雷射射出聚焦鏡之距離,以 對卵細胞進行穿孔、去核,以及牽引體細胞並將其注射進 , 入卵細胞之操作,最後以雷射光學融合或以化學激活,而 達到自動化操作細胞核之功效者。 【實施方式】 - 為使貴審查委員對本案所述細胞核操作的自動化裝置 的結構特徵與功效,能有更進一步的認識與瞭解,茲舉一 些可行實施例並配合圖示,詳細說·明如下: 首先請參閱第一圖至第四圖所示,本創作「細胞核操 作的自動化裝置」主要構造包括由一自動化操作運動單元 + 10、一三向自由度運動單元20、一雙軸顯微鏡單元30、一 雷射光機電單元40及一電腦控制單元50所組成;其特徵 在: C8)Page 6 M290505 Fourth, the creation instructions (3) is fixed on the traverse mechanism of the automated operation of the motion unit corresponding to the vertical video microscope from the bottom up, and coaxial with the vertical video display mirror; computer control unit is used Connect with other unit signals to control the automatic carrier of each unit. When the device is used as described above, a microfluidic slide is used to load the microfluids of the egg cells, the somatic cells and the culture solution, and the plurality of slides are placed in the three-dimensional degree of freedom movement unit. Automated operation of the motion unit on the glass table, and then use the vertical and horizontal video microscope of the dual-axis microscope unit to capture the XY and XZ plane images of the egg cells in the microfluidics, transmit them to the computer control unit to display their images and calculate the egg cells. The elevation position of the z-axis, and then the laser electromechanical unit disposed directly below the vertical microscope, and the computer automatically adjusts the distance of the laser to emit the focusing mirror to perforate and enucle the egg cells, and to pull the body cells and inject them into the z-axis. , the operation of the egg cells, and finally by laser optical fusion or chemical activation, to achieve the function of automated operation of the nucleus. [Embodiment] - In order to enable the reviewing committee to have a better understanding and understanding of the structural features and effects of the automated device for the operation of the cell core described in this case, some feasible embodiments are shown in conjunction with the drawings, and the details are as follows: : First, please refer to the first to fourth figures. The main structure of the "automated device for nuclear operation" includes an automated operation unit + 10, a three-dimensional degree of freedom motion unit 20, and a dual-axis microscope unit 30. , a laser electromechanical unit 40 and a computer control unit 50; characterized by: C8)
第7頁 M290505Page 7 M290505
機構1一2及一具有載玻台i 4之縱移機構丨3 (如第==橫移 三向自由度運動單元20包含一吸盤式機 f曰丄 向線性滑軌2 2 (如第一圖),而可將在待料谇育^的、 玻片24吸起移至自動化操作運動單元1〇的載^台“,再由 ί ΐ 操作後之玻片24移至成品培育箱25存放, 七#片/的上表面具有一橫向的微流體241 ,該微 =,L體 見度及深度略大於卵細胞之直徑(如第四圖The mechanism 1 - 2 and a vertical displacement mechanism 具有 3 having a glass table i 4 (such as the == traverse three-dimensional degree of freedom motion unit 20 includes a suction cup machine f 线性 linear slide 2 2 (such as the first Fig.), the slide 24 can be moved up to the loading table of the automatic operation movement unit 1", and then the slide 24 after the operation is moved to the finished incubator 25 for storage. , the upper surface of the seven #片/ has a lateral microfluidic 241, the micro =, L body visibility and depth is slightly larger than the diameter of the egg cell (such as the fourth figure
蒋機n η ί元30係固定在自動操作運動單元10之橫 ,鏡32及一側向螢光燈33組成,該二顯微鏡31與32互j9〇 度的排列且鏡頭對向載玻台14的位置(如第二圖),且每 一顯微鏡31與32均包含有一CCD攝影機31 i、321及一同軸 射出雷射二極體312、322 ;The Jiang machine n η ί元 30 series is fixed on the horizontal direction of the automatic operation motion unit 10, and the mirror 32 and the side are arranged to the fluorescent lamp 33. The two microscopes 31 and 32 are arranged in a j9 degree and the lens is opposed to the glass table 14 Position (as shown in the second figure), and each of the microscopes 31 and 32 includes a CCD camera 31 i, 321 and a coaxial injection laser diode 312, 322;
/雷射光機電單元4〇其至少包含有一雷射二極體41、一 f ί,射機42及一射出聚焦鏡43,該雷射二極體41與脈衝 苗射機42係固定於自動化操作運動單元1 0之機架1 1底部, =可經同一路控傳送至射出聚焦鏡4 3後向上照射;該射出 水:鏡4 3則係固定於自動化操作運動單元1 〇之橫移機構1 2 上對應於垂直視頻顯微鏡31下方由下向上照射,且與垂直 ,頻顯微鏡3 1同軸(如第二、第四及第五圖)·,又在本實 ^例中’ 5玄射出聚焦鏡4 3係固定在一垂直升降座& 4上,,可 藉以調整雷射聚焦點的高度位置;The laser electromechanical unit 4 includes at least one laser diode 41, a f ί, a ejector 42 and an emission focusing mirror 43. The laser diode 41 and the pulse ejector 42 are fixed in an automated operation. The bottom of the frame 1 1 of the moving unit 10, = can be transmitted to the exit focusing mirror 43 by the same way, and then irradiated upward; the injected water: the mirror 4 is fixed to the traverse mechanism 1 of the automatic operation movement unit 1 2 corresponds to the vertical video microscope 31 from the bottom to the top, and is perpendicular to the vertical, frequency microscope 3 1 (such as the second, fourth and fifth maps) · In this example, '5 Xuanguang focusing mirror 4 3 is fixed on a vertical lifting seat & 4, which can be used to adjust the height position of the laser focusing point;
M290505M290505
電腦控制f元5〇孫用來 以控制各單元自動運作者。 單元線 如上所述裝置使用時,係將含有卵細胞6 〇及體細胞之 培養液載於玻片24之微流體241之中,即可利用玻片之微 流體241的寬度略為固定卵細胞6〇在玻片以上之位置,二 後由三向自由度運動單元20將數個玻片24橫列的放置於、、一 I 呆Γ運ί單元1〇之載玻台14上,#藉橫移機構12移 又車颁微鏡單元3 0的位置,及藉縱移機構丨3移動載玻台Computer control f yuan 5 grandson is used to control the automatic carrier of each unit. When the unit line is used as described above, the culture medium containing the egg cells and the somatic cells is placed in the microfluidic 241 of the slide 24, and the width of the microfluidic 241 of the slide is used to slightly fix the egg cells. Above the slide, after the three-way degree of freedom movement unit 20, a plurality of slides 24 are placed on the glass table 14 of the I, the I squatting unit, and the traverse mechanism 12 shifting and giving the position of the micromirror unit 30, and moving the glass table by the vertical shifting mechanism 丨3
的位置,使二顯微鏡3丨、3 2的鏡頭可對準玻片2 4上所載 細胞60,再利用雙軸顯微鏡單元3〇的垂直及水平視頻 鏡31、32擷取在微流體241内卵細胞之^與^的 影像(如第六圖)。 如上述’由雙軸顯微鏡單元3 〇所擷取之卵細胞在χγ與 的平面影像資料傳輸到電腦控制單元5 〇之後,其可將影 像顯示出來,並利用與二顯微鏡3丨、32同軸射出之雷射光 $側向螢光燈3 3照射出之螢光激發光源的作用,而產生的 :1位對比現象計,以計算出卵細胞6 〇在Ζ軸方向的正確 咼5位置,另,利用設置於垂直顯微鏡3 1正下方同步移動 之雷$射出聚焦鏡4 3,配合電腦控制單元5 〇自動調整雷射 $出聚焦鏡43之距離,以雷射二極體41發出之雷射光將前 述之卵細胞定位,再以脈衝雷射機42發射 雷射光進行對 ,細胞進行m、去核,以及牵引體細胞並;^射進入 印細胞之操作’最後再以雷射光學融合或以化學激活,而 達到自動化操作細胞核之功效者。The position of the two microscopes 3, 3 2 can be aligned with the cells 60 carried on the slides 24, and then captured in the microfluidics 241 using the vertical and horizontal video mirrors 31, 32 of the biaxial microscope unit 3 An image of the ^ and ^ of the egg cell (as in Figure 6). After the above-mentioned egg cells extracted by the two-axis microscope unit 3 are transferred to the computer control unit 5 χ, the image data of the χγ and the 控制γ can be displayed, and the image is generated coaxially with the two microscopes 3, 32. Laser light $ lateral fluorescent lamp 3 3 illuminates the role of the fluorescent excitation light source, and produces: 1 position contrast phenomenon meter to calculate the correct 咼 5 position of the egg cell 6 〇 in the x-axis direction, and Synchronously moving the laser to the vertical microscope 3 1 and shooting the focusing mirror 4 3, and automatically adjusting the distance of the laser focusing mirror 43 with the computer control unit 5 ,, the laser light emitted by the laser diode 41 will be the aforementioned The egg cells are positioned, and then the laser light is emitted by the pulse laser device 42 to perform m, enucleation, and pulling the body cells; and the operation of entering the cells is finally laser-polarized or chemically activated. Achieve the ability to automate the operation of the nucleus.
(s) M290505 四、創作說明(6) 綜上所述,本創作所揭露的是一種細胞核操作的自動 化裝置,其確實能達到即時監控及自動操作之功效,並能 快速且穩定的確認卵細胞之位置,達到使自動化操作細胞 核之功效;又,本案所述之裝置及其特徵在提出專利申請 前未見於刊物或公開使用者;故本創作能符合新型專利要 件,爰依法申請新型專利。(s) M290505 IV. Creation Instructions (6) In summary, this creation reveals an automated device for nuclear operation that can achieve immediate monitoring and automatic operation, and can quickly and stably confirm egg cells. Position, to achieve the effect of automated operation of the nucleus; in addition, the device and its features described in this case were not found in the publication or public users before the patent application; therefore, the creation can meet the new patent requirements, and apply for a new patent according to law.
第10頁 M290505 «式簡單說明 第一圖 W%創作之餐體裝置立體示意圖 第二圖 係本創作之側視平面組合圖 第三圖 係本創作之自動化操作運動單元示意圖 第四圖 係本創作之玻片示意圖 第五圖 係本創作之雷射光機電單元的光路示意圖 第六圖 係本創作之雙軸顯微鏡單元取像示意圖(其中之 玻片及卵細胞的比例放大示意) 【主要元件符號說明】 自動化操作運動單元10 三向自由度運動單元20 雙軸顯微鏡單元 30 雷射光機電單元 40 i >電腦控制單元 50 卵細胞 60 機架 11 橫移機構 12 縱移機構 13 載玻台 14 吸盤式機械臂 2 1 橫向線性滑執 22 待料培育箱 23 玻片 24 成品培育箱 25 垂直視頻顯微鏡 31 - 水平視頻顯微鏡 32 側向螢光燈 33 雷射二極體 41 脈衝雷射機 42 射出聚焦鏡 43 垂直升降座 44 微流體 241 C C D攝影機3 1 1、 321 雷射二極體31 2、 322Page 10 M290505 «Simple description of the first picture W% created a three-dimensional diagram of the food device installation The second picture is the side view plane combination of the creation of the third picture of the creation of the automatic operation of the motion unit diagram fourth picture of the creation The fifth diagram of the slide diagram is the schematic diagram of the optical path of the laser electromechanical unit of the present invention. The sixth diagram is a schematic diagram of the biaxial microscope unit of the present invention (the scale of the slide and the egg cell is enlarged) [Main component symbol description] Automated operation of motion unit 10 Three-way freedom motion unit 20 Dual-axis microscope unit 30 Laser electromechanical unit 40 i > Computer control unit 50 Egg cell 60 Rack 11 Traverse mechanism 12 Vertical mechanism 13 Glass table 14 Suction cup arm 2 1 Horizontal linear slipper 22 Waiting incubator 23 Slide 24 Finished incubator 25 Vertical video microscope 31 - Horizontal video microscope 32 Lateral fluorescent light 33 Laser diode 41 Pulse laser 42 Out of focus mirror 43 Vertical Lifting seat 44 microfluid 241 CCD camera 3 1 1, 321 laser diode 31 2, 322
第11頁Page 11
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TWI401472B (en) * | 2008-09-13 | 2013-07-11 | Japan Science & Tech Agency | Microscope apparatus and fluorescence observing method using the same |
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TWI401472B (en) * | 2008-09-13 | 2013-07-11 | Japan Science & Tech Agency | Microscope apparatus and fluorescence observing method using the same |
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