M290504 八幹型說明4------— 【新型所屬之技術領域】 本新型疋有關於一種流式細胞儀，特別是指一種運用 於微流體晶片上，且可簡化操作程序、整合光學偵測及觀 測的儀器。 【先前技術】 近年來，由於在生物學研究中單株抗體技術及螢光染 料的發展，並結合了細胞培養方法的進步，流式細胞技術 (Flow cytometry)的發展達到了巔峰，隨之而來的就是於各 領域中的廣泛應用。而到了 90年代，流式細胞技術已經深 入到樂物學、生物學、基因體學及臨床醫學等研究領域， 藉由流式細胞技術，研究員可快速偵測細胞的表面抗原、 胞内抗原、轉殖基因的表達、細胞機能、去氧核糖核酸 (Deoxyribonucleic acid，DNA)分析、細胞凋亡分析、細胞週 期分析、細胞活性分析、自由基測量、細菌增殖曲線等等 ，此外，更可利用細胞分選（Cell sorting)功能，將特定目標 之細胞分選出來，可重新回收利用特殊細胞或細菌顆粒族 群，進一步培養或選殖，因此流式細胞儀成為應用於免疫 學，微生物學，細胞學等各項實驗之重要儀器。 另一方面，由於源自於半導體製程技術之微機電系、、统 (Micro-Electro-Mechanical System，MEMS)科技近年來的迅 速發展，舉凡在各領域中皆有許多嶄新的突破，並應用其^ 各種創新之製程技術，可精密的製作出各式各樣的微型元 件。而將微機電系統概念導入醫學分析之領域，衍生出微 M290504 小化快速之女，將^型生化分育 儀器縮小並整合至一微小的生物晶片中，可有效將進樣、 反應、分離、偵測及分析等操作步驟整合於一微型晶片， 並具有體積小、分析樣本需求少、反應時間快及頻率響應 间、檢測精確度高等優點，再結合可大量批次製造之優點 ’因此可達到提升量測之靈敏度、減少生物試劑耗量及儀 器成本的效果，進而達到晶片實驗室（Lab_〇n_a_chip，L〇c) 之目標。 在大型流式細胞儀設備中，普遍使用光學偵測方式來 完成檢體的測量及分析，而細胞樣本流（§&1111)16 fl〇w)首先由 兩侧的邊鞘流（Sheath flow)所產生水力聚焦（Hydr〇dynamic focusing)方式聚焦，並通過入射之雷射光束，藉由雷射光束 對於細胞及生物分子樣本所產生之折射及反射現象分析細 胞的大小及種類，更可利用螢光標定（Flu〇rescence 1^^叫） 之方式’達成分析及分類不同細胞樣本之效果。然而，在 大型流式細胞儀系統中，仍需使用高價精密的光學偵測系 統，不但價格昂貴，使用時也需經過繁雜的光學校準程序 並且而要經由训練合格的使用者來進行操作，因此多為 一般醫院及教學研究單位使用，普及率始終無法提高。 然而，應用微;f幾電技術所製作之微型力式細胞分析晶 片可有效的縮小分析系統尺寸及降低價格，改善大型系統 高價及操作繁雜之缺點，且在有效的縮小體積後，更容易 被個域用者所接受。由於微型化流式細胞分析晶片技術 具有高解析度及靈敏度之優點，近年來被各國研究團隊廣 M290504 泛研究^ 材^金屬、石夕、玻璃及塑膠高分子等材料。然而卻缺乏一 H化&作程序、整合光學制及觀測的儀器。 【新型内容】 敫八、^ $本新型之目的，即在提供一種可簡化操作程序 、整合光學偵測及觀測的偵測微流體晶片之流式細胞儀。 本新型之另一目的，即在提供一種體積小、成本低的 福測微流體晶片 < 流式細胞儀。 曰於是’本新型偵測微流體晶片之流式細胞儀，該微流 體曰曰片^有一樣品流、兩邊鞠流及一摘測區段，樣品流是 位於兩邊鞘流間，並於同一處連通於偵測區段。該流式細 胞:義包括有-控制裝置、一趨動裝置、一光源裝置、一伯 ^ 取像裝置。趨動裝置是用以驅動樣品流及邊鞘 "'光源破置則可發出一光源投射在偵測區段，使流經偵 [”又的、、、田胞被激發出特定波長的螢光，該偵測裝置是設 置於忒螢光的路徑上，用以偵測該螢光，取像裝置則是用 以拍攝出細胞於此過程中的影像。 進步的包括有一殼體，將該等裝置整合於殼體内， 縮小整體體積。 更進一步的可將偵測裝置及取像裝置的輸出整合於殼 體上的一顯示器。 該光源裳置包括有一雷射光源。 該光源衣置包括有一發光二極體（Light ^ Di〇de ，LED)光源。 M290504 4取像裝置包括有一電荷_合元件（C^Weoupkd Device，· CCD)攝影機。 【實施方式】 有關本新里之别述及其他技術内容、特點與功效，在 以下配口參考圖式之較佳實施例的詳細說明中，將可清楚 的呈現。 、如圖1、2、3所不，為本新型一較佳實施例流式細胞 儀100，疋配合一微流體晶片900使用，該微流體晶片9〇〇 具有一樣品流91、兩邊鞘流92及一偵測區段93，樣品流 91疋位於兩邊鞘流92間，且樣品流91之聚焦是利用兩邊 勒机92與樣品流91流速的不同，而產生之水力集中 (Hydrodynamics f0CUSing)現象。由兩側邊鞘流92對中間樣 口口 "IL 91的水力壓縮，使樣品流91的寬度於偵測區段93縮 減至預定之尺寸，利用此原理可將樣品流9丨中之細胞檢體 被擠壓，並於偵測區段93排列為一直線。 該流式細胞儀100包括有一殼體20、一控制裝置30、 一趨動裝置40、一光源裝置50、一偵測裝置6〇及一取像 裝置70。 该殼體20呈一矩形且為上開放式的，所以具有一容置 空間21，於殼體20其中一側邊樞接有一蓋體23，容置空 間21與蓋體23間設有一隔架25，另於容置空間21的一側 壁貫穿有一晶片插置槽27，於相鄰的另一側壁對應的設有 一定位導軌28，所以微流體晶片900可由晶片插置槽27插 M290504 入，並·^------------ 該控制裝置30是設於殼體20隔架25的下方，並具有 一設置在隔架25上的鍵盤31及一第一顯示器33,本實施 例的第一顯示器33為一般傳統的16χ2點矩陣型LCD顯示 器，用以顯示鍵盤3 1輸入的數值資料。 趨動裝置40是組裝於容置空間21内，其是用來與微 流體晶片900配合，驅動微流體晶片9〇〇中的樣品流91及 兩邊鞘流92。 由於本貝加例的微流體晶片900是以氣動式微型幫浦 來達到驅動微流體的目的，因此本實施例的趨動裝置4〇具 有一氣壓源41、複數個電磁閥43及複數個管路45，本實 她例中的氣壓源41為一種内裝高壓氣體的小鋼瓶，其可輸 出固定氣壓值的氣體，該等管路45是與氣壓源41相互連 通’且每-管路45搭配有-電磁閥43，本實施例的電磁閱 43及管路45是經過整合的，所以於圖中並未看到複數個， 而是將所有電磁閥43整合於一盒體内，將所有管路45整 合於一較大管子内。 本實施例的趨動裝置40更包括一樞接於殼體2〇隔架 25上的氣壓板47,氣壓板47是樞設於對應晶片插置槽π 及定位導軌28處’該等管路45是延伸至氣壓板47,並分 別於氣壓板47朝向容置空間21的一表面凸伸出一插管451 ，該等插管451是對應於微流體晶片9〇〇上的相對位置。 本實施例中，藉由調整電磁閥43來達到控制管路Μ 進氣的頻率，進而驅動微流體晶片_中的樣品流91及兩 M290504 且其開關的頻率為可調式設計。本實施例電磁閥43的頻率 調整上，設計成以5的倍數為頻率調整的間距，其範圍為 5Hz〜200Hz。 趨動裝置40之氣壓源41亦可為其他形式，如空壓機 ，只要此長:供適當且固定壓力的氣體即可。 光源裝置50是設於容置㈣21内，且是對應於微流 體晶片900插入處的下方，其包括有一光源51、一第一反 射鏡53及一物鏡55，該光源51為一綠光雷射，並且配合 一帶通濾片511(Band pass，BP)將其他波長的螢光全部阻斷 ，避免產生影響，本實施例為530nm的帶通濾片，理論上 其只允許發射的530 nm綠色光通過，而其他波長的光全部 被阻斷，但一般的帶通濾片是可允許正、負1〇nm的範圍 ，所以可選用容許範圍内的帶通濾片。當然於其他實施例 亦可使用其他不同波長的雷射及帶通濾片，亦可使用適當 的LED來取代雷射光源。 本實施例的物鏡55具有十倍光學，且是對準設置於微 流體晶片900偵測區段93的下方；第一反射鏡53於本實 施例中為一 585nm長波通濾片（Long pass，LP)，其只允許大 於585nm波長的光通過，而發射的525nm綠光則全部反射 所以由光源51所發出的綠光雷射係全反射的經由物鏡 55投射在偵測區段93，細胞吸收特定波長的光能量，而被 激發出特定波長的螢光，當然細胞於進樣前必須先進行螢 光標定的動作。 10 M290504 -莽於音光標资知該項龙ir者能輕易— ’且螢光標定也會因細胞的不同而有所差異，所以以下僅 大概的說明。 以一樣本細胞來說明，將細胞培養在含10%胎牛血清 (Fetal Bovine Serum，FBS)在 DMEM(Dulbecco’s Modified Eagle Medium)的培養基下經48小時培養於溫度37 °C、5 % -^*氧化奴（C〇2)之培養箱。M290504 Eight dry type description 4------— 【New technology field】 This new type relates to a flow cytometer, especially to a microfluidic wafer, which simplifies the operation procedure and integrates optics. Instrument for detection and observation. [Prior Art] In recent years, due to the development of monoclonal antibody technology and fluorescent dyes in biological research, combined with advances in cell culture methods, the development of flow cytometry has reached its peak, followed by It comes to a wide range of applications in various fields. In the 1990s, flow cytometry has penetrated into the fields of music science, biology, genomics, and clinical medicine. With flow cytometry, researchers can quickly detect surface antigens, intracellular antigens, and Transgenic gene expression, cell function, deoxyribonucleic acid (DNA) analysis, apoptosis analysis, cell cycle analysis, cell activity analysis, free radical measurement, bacterial proliferation curve, etc. In addition, cells can be utilized The Cell sorting function sorts out specific target cells and re-uses special cells or bacterial particle populations for further culture or colonization. Therefore, flow cytometry is applied to immunology, microbiology, and cytology. An important instrument for various experiments. On the other hand, due to the rapid development of Micro-Electro-Mechanical System (MEMS) technology derived from semiconductor process technology in recent years, there are many new breakthroughs in various fields, and its application ^ A variety of innovative process technology, can produce a variety of miniature components. And the introduction of the concept of MEMS into the field of medical analysis, derived from the micro-M290504 small and rapid woman, the ^ type biochemical equipment is reduced and integrated into a tiny bio-chip, which can effectively inject, react, separate, The detection and analysis steps are integrated into a microchip, and have the advantages of small size, low sample analysis requirements, fast response time, high frequency response, high detection accuracy, and the advantages of large batch manufacturing. Improve the sensitivity of measurement, reduce the consumption of biological reagents and the cost of the instrument, and then achieve the goal of the wafer laboratory (Lab_〇n_a_chip, L〇c). In large flow cytometry equipment, optical detection is commonly used to complete the measurement and analysis of the sample, while the cell sample flow (§&1111) 16 fl〇w) is firstly carried out by the side sheath flow (Sheath flow) The hydration dynamic focusing method is used to focus and analyze the size and type of the cells by the incident laser beam and the reflection and reflection phenomenon of the laser beam on the cells and biomolecule samples. The method of 'Flu〇rescence 1^^calling' is used to analyze and classify the effects of different cell samples. However, in large flow cytometry systems, high-precision optical detection systems are still required, which are expensive, require complicated optical calibration procedures, and are operated by trained users. Therefore, it is mostly used by general hospitals and teaching research units, and the penetration rate cannot be improved. However, the application of micro-power micro-power cell analysis wafers can effectively reduce the size of the analysis system and reduce the price, improve the high cost of large systems and the complicated operation, and it is easier to be effectively reduced in size. Accepted by domain users. Due to the advantages of high resolution and sensitivity, the miniaturized flow cytometry wafer technology has been widely researched by various research teams in recent years, M290504, metal, stone, glass and plastic polymers. However, there is a lack of an instrument for integrating procedures and integrating optical systems and observations. [New Content] 本8, ^ $ The purpose of this new type is to provide a flow cytometer for detecting microfluidic wafers that simplifies the operation procedure and integrates optical detection and observation. Another object of the present invention is to provide a small-volume, low-cost measurement microfluidic wafer <flow cytometry. The present invention is a flow cytometer for detecting a microfluidic wafer. The microfluidic cymbal has a sample flow, turbulence on both sides, and a sampling section. The sample flow is located between the sheath flows on both sides, and is in the same place. Connected to the detection section. The flow cell includes: a control device, a kinetic device, a light source device, and a cymbal imaging device. The kinetic device is used to drive the sample stream and the edge sheath "'the light source is broken to emit a light source to be projected on the detection section, so that the flow field is excited, and the field cell is excited to emit a specific wavelength. Light, the detecting device is disposed on the path of the fluorescent light to detect the fluorescent light, and the image capturing device is configured to capture images of the cells in the process. The improvement includes a casing, and the The device is integrated into the housing to reduce the overall volume. Further, the output of the detecting device and the image capturing device is integrated into a display on the housing. The light source includes a laser light source. There is a light-emitting diode (LED) light source. The M290504 4 image capture device includes a C_Weoupkd Device, CCD camera. [Embodiment] Other technical contents, features, and effects will be apparent from the detailed description of the preferred embodiments of the following reference drawings, as shown in FIGS. 1, 2, and 3, which are a preferred embodiment of the present invention. Flow cytometer 100, with a microfluid The wafer 900 is used. The microfluidic wafer 9 has a sample stream 91, a sheath flow 92 and a detection section 93. The sample stream 91 is located between the sheath streams 92, and the sample stream 91 is focused by using both sides. The hydrodynamics f0CUSing phenomenon is caused by the difference between the flow rate of the machine 92 and the sample stream 91. The hydraulic compression of the intermediate sample mouth "IL 91 by the side sheath flow 92 causes the width of the sample stream 91 to be detected. The segment 93 is reduced to a predetermined size, and the cell sample in the sample stream 9 被 can be squeezed by this principle and arranged in a line in the detection section 93. The flow cytometer 100 includes a housing 20, A control device 30, a oscillating device 40, a light source device 50, a detecting device 〇, and an image capturing device 70. The housing 20 has a rectangular shape and is open-ended, so that it has an accommodating space 21 A cover body 23 is pivotally connected to one side of the housing 20, a partition 25 is disposed between the accommodating space 21 and the cover body 23, and a sidewall of the accommodating space 21 is inserted through a wafer insertion slot 27. The other side wall of the adjacent one is correspondingly provided with a positioning guide 28, so the microfluidic crystal 900 can be inserted into the chip insertion slot 27, M290504, and the control device 30 is disposed under the partition 25 of the housing 20, and has a partition 25 disposed thereon. The first display 33 of the present embodiment is a conventional 16” 2-point matrix type LCD display for displaying the numerical data input by the keyboard 31. The oscillating device 40 is assembled and received. Within space 21, it is used to cooperate with microfluidic wafer 900 to drive sample stream 91 and both sheath streams 92 in microfluidic wafer 9A. Since the microfluidic wafer 900 of the present embodiment is a pneumatic micro-pull to achieve the purpose of driving the microfluid, the oscillating device 4 of the present embodiment has an air pressure source 41, a plurality of electromagnetic valves 43, and a plurality of tubes. Road 45, in the example of the air source 41 is a small cylinder containing a high-pressure gas, which can output a gas of a fixed pressure value, the line 45 is in communication with the air pressure source 41 'and each-line 45 With the solenoid valve 43, the electromagnetic reading 43 and the line 45 of the present embodiment are integrated, so that a plurality of solenoid valves 43 are not integrated into one box, and all of them are integrated into one box. Line 45 is integrated into a larger tube. The oscillating device 40 of the present embodiment further includes a gas pressure plate 47 pivotally connected to the housing 2 〇 spacer 25, and the air pressure plate 47 is pivotally disposed at the corresponding wafer insertion groove π and the positioning rail 28 45 extends to the air plate 47, and a cannula 451 is protruded from a surface of the air plate 47 facing the accommodating space 21, respectively, and the cannula 451 corresponds to a relative position on the microfluidic chip 9 。. In this embodiment, by adjusting the solenoid valve 43 to achieve the frequency of the control line 进气 intake, the sample stream 91 and the two M290504 in the microfluidic wafer are driven and the frequency of the switch is adjustable. In the frequency adjustment of the solenoid valve 43 of the present embodiment, the pitch is adjusted at a frequency of a multiple of 5, and the range is 5 Hz to 200 Hz. The air pressure source 41 of the actuating device 40 can also be in other forms, such as an air compressor, as long as it is suitable for a suitable and fixed pressure gas. The light source device 50 is disposed in the accommodating (four) 21 and corresponding to the lower portion of the insertion of the microfluidic chip 900. The light source device 50 includes a light source 51, a first mirror 53 and an objective lens 55. The light source 51 is a green laser. And with a bandpass filter 511 (Band pass, BP) to block all other wavelengths of fluorescence to avoid the impact, this embodiment is a 530nm bandpass filter, theoretically only allowed to emit 530 nm green light Passing, while other wavelengths of light are all blocked, but the general bandpass filter is allowed to allow positive and negative 1 〇 nm range, so bandpass filters within the allowable range can be selected. Of course, other different wavelengths of laser and band pass filters may be used in other embodiments, and a suitable LED may be used instead of the laser source. The objective lens 55 of the present embodiment has ten times of optics and is disposed below the microfluidic wafer 900 detecting section 93. The first mirror 53 is a 585 nm long pass filter (Long pass, in this embodiment). LP), which only allows light having a wavelength greater than 585 nm to pass, and the emitted 525 nm green light is totally reflected. Therefore, the green light emitted by the light source 51 is totally reflected by the objective lens 55 projected on the detecting section 93, and the cell absorbs the specific The light energy of the wavelength is excited by the fluorescence of a specific wavelength. Of course, the cell must perform the action of the cursor before the injection. 10 M290504 - The sound cursor knows that the dragon ir can easily - 'and the cursor will vary from cell to cell, so the following is only a general description. In the same manner as the present cells, the cells were cultured in a medium containing 10% fetal calf serum (Fetal Bovine Serum, FBS) in DMEM (Dulbecco's Modified Eagle Medium) for 48 hours at a temperature of 37 ° C, 5% -^* Incubator for oxidized slave (C〇2).
所使用之螢光染料為MitoTracker® Red 580，使用激發 光源波長為58 lnm，散發波長為644nm，本勞光染料主要是 標記於細胞的粒線體（Mitochondria)中。細胞樣本與螢光染 料使用的標定方式為直接免疫螢光染色法，直接免疫螢光 染色法可應用於任何抗原的物質，不管在細胞内或細胞外 、原蟲、細菌、立克次氏體、病毒、組織抗原、激素或酵 素。利用此方式，將可分別針對不同之樣本做特定之螢光 標定，不同之螢光染劑可分別被不同之波長光源所激發， 利用此方式將可分辨出各個不同之檢體。 偵測裝置60是設於容置空間21内，包括有一接收器 61及第一顯不器63，另外，於前述第一反射鏡53後方 ,又置有第一反射鏡65，第二反射鏡65於本實施例中為一 全反射的鏡面。 Α接收^§ 61於本實施例中為一雪崩光電二極體，該； 一』U3為一液晶螢幕，並組設於蓋冑μ内面，接4 器61與第二顯示63 α 土The fluorescent dye used was MitoTracker® Red 580, which used an excitation source with a wavelength of 58 lnm and a emission wavelength of 644 nm. The Labo dye was mainly labeled in the cell mitochondria (Mitochondria). The calibration method for cell samples and fluorescent dyes is direct immunofluorescence staining, and direct immunofluorescence staining can be applied to any antigen, whether intracellular or extracellular, protozoa, bacteria, rickettsia. , viruses, tissue antigens, hormones or enzymes. In this way, specific fluorescent calibrations can be performed for different samples, and different fluorescent dyes can be excited by different wavelength sources respectively, and different methods can be distinguished by this method. The detecting device 60 is disposed in the accommodating space 21 and includes a receiver 61 and a first display 63. Further, behind the first mirror 53, a first mirror 65 and a second mirror are disposed. 65 is a totally reflective mirror in this embodiment. ΑReceiving ^§ 61 is an avalanche photodiode in this embodiment, and the U3 is a liquid crystal screen, and is disposed on the inner surface of the cover ,μ, connected to the device 61 and the second display 63 α
均電性連接於控制裝置30，接收I 61用以偵測由第二及射倍Α 一反射鏡65反射過來的光學訊號，控制^ M290504 _要一換责意以電位脈衝型式顯示 於第二顯示器63上，本實施例於第二反射鏡65與接收器 61間設有一 647nm帶通濾片67，所以接收器61只會接收 到647nm正、負10ηιη波長的光；而於帶通濾片67與第二 反射鏡65間設有一十倍光學的目鏡68，增加訊號強度。 該取像裝置70是設於容置空間21内，包括有一攝影 機71及一第三顯示器73，只是本實施例的第三顯示器73 即為第一顯示器63，也就是說彳貞測裝置60與取像裝置70 ► 共用一顯示器，以下僅以第二顯示器63來說明。另外，亦 可進一步的將前述第一顯示器33 —起整合，也就是說第一 、笫一、弟二顯示器33、63、73為同一顯示器，可將空間 做其他運用，例如供一些必要用具或檢測液體置放等。 攝影機71為一 CCD型式的攝影機，且電性連接於控制 裝置30，係用以拍攝由第二反射鏡65反射過來的影像訊號 ，控制裝置30再將影像訊號顯示於第二顯示器63上，所 • 以第二顯示器63是可供切換的顯示電位脈衝或影像訊號。 然而，本實施例為了使接收器61與攝影機71可以同 時接收§fl號，所以於647nm帶通濾片67後方增設有一 30/70的分光濾波片75,也就是7〇百分比的光會穿透而供 接收器61接收，30百分比的光會被反射至攝影機71，供 攝影機71接收，所以於硬體裝置上無需移動接收器6丨與 攝影機71，即可於第二顯示器63上切換觀察電位脈衝或影 像訊號。 紅上所述，當欲偵测一細胞時，係先將細胞樣本於進 12 M290504 — ~~樣^^行螢光標定，—之一後將加胞樣本置贫贫流體晶片一 樣品流91，並以純水或其他不影響偵測的其他適當液體置 於兩邊鞘流92，然後將微流體晶片900由晶片插置槽27插 入’沿著定位導軌2 8進入容置空間21。 然後將氣壓板47朝向容置空間21蓋合，使該等插管 45 1對應的插置於微流體晶片900上的相對位置，啟動趨動 裝置40驅動微流體晶片900中的樣品流91及兩邊鞘流92 I ，使樣品流91與兩邊鞘流92利用流速的不同聚焦，將樣 π口流91中之細胞檢體被擠壓，並於偵測區段93排列為一 直線。 啟動光源裝置50、偵測裝置60及取像裝置7〇，光源 5!發射出綠光雷射，經由第一反射鏡53反射，再經由物鏡 55投射在偵測區段93，細胞檢體吸收特定波長的光能量， 而被激發出特定波長的螢光，該螢光波長是大於第一反射 鏡53的585nm，所以會穿透第一反射鏡53。 • 再由第二反射鏡65的反射，經過十倍光學的目鏡68 及647nm帶通濾片67,之後經過3〇/7〇的分光濾波片乃， 供接收器61及攝影機71接收，即可於第二顯示器63上切 換觀察電位脈衝或影像訊號。而若偵測裝置6〇及取像裝置 7〇非共用同-顯示器的話，即可同時觀察電位脈衝及影像 訊號。 。如圖4為以前述的樣本細胞來實驗所得的電位脈衝訊 唬圖’圖中X軸為債測時間’ γ轴為電壓值，圖中的雜訊 基準值為UmV，而每個減峰值為經過螢光標定之細胞樣 13 M290504 激發光爺激發所產i之螢光訊號------ 歸納上述，本新型之流式細胞儀，可同時觀察電位脈衝 及影像訊號，且全程的操作簡易，儀器的結構成本相較於 習知大型的流式細胞儀所使用的光電倍增管（PMT)比較，本 新型的價格及體積都大為減少。 惟以上所述者，僅為本新型之較佳實施例而已，當不 能以此限定本新型實施之範圍，即大凡依本新型申請專利 範圍及新型說明内容所作之簡單的等效變化與修飾，皆仍 屬本新型專利涵蓋之範圍内。 【圖式簡單說明】 圖1是本新型較佳實施例的立體示意圖； 圖2是圖1較佳實施例另一狀態的立體示意圖，圖中 爲了便於觀察容置空間，所以殼體的蓋體及隔架並未繪出 圖3是圖1較佳實施例架構示意圖，及光學路徑示音 圖；及 ^ 圖4是以一樣本細胞來實驗所得的電位脈衝訊號圖。 14 M290504The power is connected to the control device 30, and the receiving I 61 is used to detect the optical signal reflected by the second and the double reflection mirror 65. The control ^ M290504 _ is replaced by a potential pulse type in the second On the display 63, in this embodiment, a 647 nm band pass filter 67 is disposed between the second mirror 65 and the receiver 61, so that the receiver 61 receives only 647 nm positive and negative 10 nm light wavelengths; and the band pass filter A ten-fold optical eyepiece 68 is disposed between the 67 and the second mirror 65 to increase the signal intensity. The image capturing device 70 is disposed in the accommodating space 21 and includes a camera 71 and a third display 73. The third display 73 of the embodiment is the first display 63, that is, the detecting device 60 and The image capturing device 70 ► shares a display, and only the second display 63 will be described below. In addition, the first display 33 can be further integrated, that is, the first, second, and second displays 33, 63, and 73 are the same display, and the space can be used for other purposes, such as providing necessary tools or Check the liquid placement and so on. The camera 71 is a CCD type camera, and is electrically connected to the control device 30 for capturing the image signal reflected by the second mirror 65. The control device 30 displays the image signal on the second display 63. • The second display 63 is a display potential pulse or video signal that can be switched. However, in this embodiment, in order to enable the receiver 61 and the camera 71 to receive the §fl number at the same time, a 30/70 spectral filter 75 is added behind the 647 nm bandpass filter 67, that is, 7% of the light will penetrate. While receiving by the receiver 61, 30% of the light is reflected to the camera 71 for the camera 71 to receive, so that the observation potential can be switched on the second display 63 without moving the receiver 6丨 and the camera 71 on the hardware device. Pulse or video signal. In the case of red, when a cell is to be detected, the cell sample is first placed in a 12 M290504-like sample, and then the cell sample is placed on a lean fluid wafer-sample stream 91. And placing the pure fluid or other suitable liquid that does not affect the detection on both sides of the sheath flow 92, and then inserting the microfluidic wafer 900 from the wafer insertion groove 27 into the accommodating space 21 along the positioning guide 28. Then, the air pressure plate 47 is covered toward the accommodating space 21, so that the corresponding insertion holes 45 1 are inserted into the relative positions on the microfluidic chip 900, and the actuating device 40 is activated to drive the sample flow 91 in the microfluidic chip 900 and The sheath flow 92 I on both sides causes the sample stream 91 and the sheath flow 92 on both sides to be focused by different flow rates, and the cell samples in the sample π mouth stream 91 are squeezed and arranged in a line in the detection section 93. The light source device 50, the detecting device 60 and the image capturing device 7 are activated, the light source 5! emits a green laser, is reflected by the first mirror 53, and is projected on the detecting section 93 via the objective lens 55, and the cell sample is absorbed. The light energy of a specific wavelength is excited by fluorescence of a specific wavelength which is larger than 585 nm of the first mirror 53, so that it penetrates the first mirror 53. • The reflection from the second mirror 65 passes through the ten-fold optical eyepiece 68 and the 647 nm band-pass filter 67, and then passes through the 3〇/7〇 spectral filter for reception by the receiver 61 and the camera 71. The observation potential pulse or image signal is switched on the second display 63. If the detecting device 6 and the image capturing device 7 are not shared, the potential pulse and the image signal can be observed at the same time. . Figure 4 shows the potential pulse signal obtained by the above sample cells. The X-axis is the debt measurement time. The γ-axis is the voltage value. The noise reference value in the figure is UmV, and each peak value is After the fluorescence of the cursor, the cell-like sample 13 M290504 activates the fluorescent signal generated by the illuminating light. In summary, the flow cytometer of the present invention can simultaneously observe the potential pulse and the image signal, and the whole process is operated. Simple, the structure cost of the instrument is greatly reduced compared to the photomultiplier tube (PMT) used in the conventional large-scale flow cytometer. However, the above description is only a preferred embodiment of the present invention, and the scope of the present invention cannot be limited thereto, that is, the simple equivalent change and modification made by the novel patent application scope and the novel description content, All remain within the scope of this new patent. BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a perspective view of a preferred embodiment of the present invention; FIG. 2 is a perspective view of another embodiment of the preferred embodiment of FIG. 1, in order to facilitate the viewing of the accommodation space, the cover of the housing 3 is a schematic view of the structure of the preferred embodiment of FIG. 1, and an optical path diagram; and FIG. 4 is a potential pulse signal obtained by experimenting with the same cell. 14 M290504
【主要元件符號說明j 100 流式細胞儀 20 殼體 21 容置空間 23 蓋體 25 隔架 27 晶片插置槽 28 定位導執 30 控制裝置 31 鍵盤 33 第一顯示器 40 趨動裝置 41 氣壓源 43 電磁閥 45 管路 451 插管 47 氣壓板 50 光源裝置 51 光源 511 帶通濾片 53 第一反射鏡 55 物鏡 60 偵測裝置 61 接收器 63 第二顯示器 65 第二反射鏡 67 帶通濾片 68 目鏡 70 取像裝置 71 攝影機 73 第三顯示器 75 分光濾波片 900 微流體晶片 91 樣品流 92 邊鞘流 93 偵測區段 15[Main component symbol description j 100 Flow cytometer 20 Housing 21 accommodating space 23 Cover body 25 Spacer 27 Wafer insertion groove 28 Positioning guide 30 Control device 31 Keyboard 33 First display 40 Actuating device 41 Air pressure source 43 Solenoid valve 45 Line 451 Cannula 47 Air plate 50 Light source device 51 Light source 511 Bandpass filter 53 First mirror 55 Objective lens 60 Detection device 61 Receiver 63 Second display 65 Second mirror 67 Bandpass filter 68 Eyepiece 70 Image capture device 71 Camera 73 Third display 75 Spectroscopic filter 900 Microfluidic wafer 91 Sample stream 92 Side sheath flow 93 Detection section 15