200940987 九、發明說明： 【發明所屬之技術領域】 -種分選有關，並且特別地，本發明係關於 種—立子分選晶片系統及其操作方法。 【先前技術】 細胞為構成生物體之最小單仅，人 類之 ❹ 胞究i臨床應用上之重要價值。舉例而言’幹細 相當i要種的細胞’對於未來細胞治療的發展 ，延些細胞的數量通常报稀少，因此，這此 • 干見細胞的取得相當不容易。 — cvtoilt統、上所!，之細胞f_義11，例如流式細胞儀(fl〇w 篩、壁之Γ0的Γ僅價格昂貴、體積魔大、需外加高電壓外，可 度還有其限制，因此並不適用於罕見細胞之篩 觸细旳叮ί i傳統上的磁珠式細胞分選儀則必須將磁珠接 =胞會汗染細胞，費用上_不如流式細胞儀如此 〇 p貝，但磁珠與儀器本身價位仍然不低。 ㈣ί外i由於流式細胞儀係使用儀11本身喊道進行細胞 之24雨後批的不同細胞樣本都是經過同一個流道，彼此 能會產生交互污染之現象，因此，對於將流式細 險。師k出的細胞應用於細胞治療上存在著不小的誤判風 曰ΰ t ’本發明之主要範•在於提供—種微流體粒子分選 曰曰片系、、先及其操作方法，以解決上述問題。 【發明内容】 200940987 根據本發日狀-具體實施例為 系統。該微流體粒子分選晶片孕士日粒子刀砥晶片 及-控制裝置。-_組及；亡、匕:偵測裝置 該微流通道包含-伽m域虚晶片上’ 判斷通過該制區域之—微粒子是====置用以 该目標微粒子移動至該目標區域。 乍用力以將 ❹ ❹ W在=應，’該微粒子可以是—細胞、—血H 頌或一磁珠。流經該微流通道的^ ^ 囷 粒子可各自依序通過該偵測ϋ稷數個_子中之每一個微 根據本發明之另-具體實施例為 片系統之操作方法。該方法係用以晶 -微流通道之一晶片。該極= 結果献，該方法控繼電極組產ΐ」 將該目;^被粒子移動至該目標區域。 並且發明之電極組及微流通道係整合於晶片中， 产體粒胞樣本可採用不同的晶片，因此，上述微 =2;=:免=上冗用之流式細胞儀容 見:r，，本發明之微流體板=== 具有極大之;^師選上，對於未來細胞治療領域之研究發展 200940987 關於本發明之優點與精神可以藉由以下的發明詳述及所 附圖式得到進一步的瞭解。 【實施方式】 根據本發明之一具體實施例為一種微流體粒子分選晶片 系統。顧名思義，該微流體粒子分選晶片系統之功用在於分 選微粒子。實際上，此處所指之微粒子可以是細胞、i球、 菌類或磁珠。於此實施例中，該微流體粒子分選晶片系統可 自一群混合的細胞中分選出罕見細胞，並避免於細胞分選過 ❹ 程中造成細胞樣本之交互污染，以利後續的細胞培養及分析 之程序。 • 請參照圖一，圖一係繪示該微流體粒子分選晶片系統之 功能方塊圖。如圖一所示，微流體粒子分選晶片系統丨包含 一晶片10、一偵測裝置12及一控制裝置14。接下來，將分 別就微流體粒子分選晶片系統1包含之各項裝置所具有的功 能進行詳細之介紹。 首先’就晶片10而於晶片10上整合有·一電極紐_ ❹ 102及一微流通道1〇4。其中微流通道1〇4包含一偵測區域 1042與一目標區域1〇44。 在實際應用中’晶片10可以是一細胞分選晶片。其主要 原理係藉由已發展成熟的微機電技術將微流道系統及電子系 統整合於微小的細胞分選晶片上，使其能真正達到可抛棄 式、個人化之目的’以減少不同的細胞樣本交互污染之風 險。 在本實施例中，微流體粒子分選晶片系統1係屬於一種 螢光激發細胞分選器（Fluorescence Activated Cell Sorter FACS) ’亦即微流體粒子分選晶片系統1將會對於通過晶片 200940987 10之偵測區域1042的每一個細胞進行螢光偵測以判斷其是 否為目標細胞。實務上，水力聚焦(hydrodynamic focusing)的 機制可被用以形成細胞樣本在晶片10之微流通道104中流動 的軌道。 於圖·一(A)中’目標細胞係以内部繪有斜線的圓圈表示， 其他細胞則以内部空白的圓圈表示。如圖二(A)所示，微流通 道104可包含三個入口 ’由中央入口流入的流體為樣本流 (central flow) 22，而從左右兩侧入口流入的流體則為邊鞘流 (sheath flow) 20。在此實施例中，中央入口之寬度為4〇微 曝米，左右兩侧入口之寬度則為400微米。根據水力聚焦原 理，位於兩側的邊鞘流20可將位於中間的細胞樣本流22夾 擠成一個細長的中央輸送流24 ’使得在中央輸送流24内的 所有細胞均能一個接一個地依序流經微流通道1〇4之偵測區 域1042。藉由此一流道之設計，不僅有助於後續細胞分選程 序之進行，亦可避免由於細胞堆積在一起而造成微流通道 104阻塞之現象發生。 接者，將就偵測裝置12進行詳細之介紹。彳貞測裝置I〕 〇 之，用在於判斷通過微流通道104之偵測區域1042的一微粒 子=否，一目標微粒子。在此實施例中，由於微流體粒子分 選晶片系統1為螢光激發細胞分選器，故偵測裝置12可藉由 螢光偵測之方式判斷通過偵測區域1〇42之細胞是否為已經 螢光標定的細胞。200940987 IX. INSTRUCTIONS: [Technical field to which the invention pertains] - Sorting is relevant, and in particular, the present invention relates to a seed sorting wafer system and a method of operating the same. [Prior Art] Cells are the smallest single species that constitute a living organism, and the human cytoplasm is of great value in clinical application. For example, 'dry and fine cells that are to be planted' are expected to be rare for the development of cell therapy in the future. Therefore, it is not easy to obtain stem cells. — cvtoilt system, 上所!, the cell f_义11, such as flow cytometry (fl〇w sieve, wall Γ 0 Γ Γ only expensive, large volume, need to add high voltage, but also its Restricted, so it is not suitable for the screening of rare cells. The traditional magnetic bead cell sorter must connect the magnetic beads to the cells, and the cost is not as good as the flow cytometry. p shell, but the price of the magnetic beads and the instrument itself is still not low. (4) ί 外 i Because the flow cytometer uses the instrument 11 itself to scream for the cells after the rain, the batch of different cell samples are all through the same flow channel, each can The phenomenon of cross-contamination is generated. Therefore, there is no small misjudgment for applying the cells of the division to the cell therapy. The main paradigm of the present invention is to provide a microfluidic particle. The invention relates to the above-mentioned problem. [Description of the Invention] 200940987 According to the present invention, the specific embodiment is a system. The microfluidic particle sorting wafer is a pregnant day particle chip and a wafer - control device. -_ group and; : detecting device, the microfluidic channel comprises - gamma m domain on the virtual wafer 'determining through the region - the microparticle is ==== is used to move the target microparticle to the target region. 乍 force to ❹ ❹ W = should, 'The microparticles can be - cells, - blood H 颂 or a magnetic bead. ^ ^ 囷 particles flowing through the microfluidic channel can each pass through the detection of each of the _ sub Another embodiment according to the present invention is a method of operating a sheet system. The method is used for a wafer of a crystal-microfluidic channel. The pole = the result, the method controls the electrode group to produce the crucible; The particles are moved to the target area. The invented electrode group and the microfluidic channel are integrated into the wafer, and the granular sample of the product can be different wafers. Therefore, the above micro = 2; =: free = redundant flow The cytometer can be seen as: r, the microfluidic plate of the present invention === is extremely great; ^ teacher selection, research development in the field of future cell therapy 200940987 The advantages and spirit of the present invention can be detailed by the following invention And the drawings are further understood. According to one embodiment of the present invention, a microfluidic particle sorting wafer system. As the name suggests, the microfluidic particle sorting wafer system functions to sort fine particles. In fact, the microparticles referred to herein may be cells, i spheres, Fungus or magnetic beads. In this embodiment, the microfluidic particle sorting wafer system can sort out rare cells from a group of mixed cells and avoid cross-contamination of cell samples during cell sorting. Subsequent procedures for cell culture and analysis. • Referring to Figure 1, Figure 1 is a functional block diagram of the microfluidic particle sorting wafer system. As shown in Figure 1, the microfluidic particle sorting wafer system includes a wafer. 10. A detecting device 12 and a control device 14. Next, the functions of each device included in the microfluidic particle sorting wafer system 1 will be described in detail. First, an electrode _ 102 and a microfluid channel 1 〇 4 are integrated on the wafer 10 for the wafer 10. The microfluidic channel 1〇4 includes a detection area 1042 and a target area 1〇44. In practical applications, the wafer 10 can be a cell sorting wafer. The main principle is to integrate the micro-channel system and the electronic system on the tiny cell sorting wafer by the well-developed micro-electromechanical technology, so that it can truly achieve the purpose of disposable and personalization to reduce different cells. The risk of sample cross contamination. In this embodiment, the microfluidic particle sorting wafer system 1 belongs to a Fluorescence Activated Cell Sorter FACS. That is, the microfluidic particle sorting wafer system 1 will pass through the wafer 200940987 10 Each cell of the detection area 1042 is subjected to fluorescence detection to determine whether it is a target cell. In practice, the mechanism of hydrodynamic focusing can be used to form a trajectory of cell samples flowing in the microfluidic channel 104 of the wafer 10. In Fig. 1 (A), the target cell line is indicated by a circle with a diagonal line inside, and the other cells are represented by a circle with an internal blank. As shown in Fig. 2(A), the microfluidic channel 104 may include three inlets 'the fluid flowing in from the central inlet is a central flow 22, and the fluid flowing in from the left and right inlets is a sheath flow (sheath) Flow) 20. In this embodiment, the central inlet has a width of 4 micrometers and the left and right inlets have a width of 400 micrometers. According to the hydraulic focusing principle, the side sheath flow 20 on either side can pinch the intermediate cell sample stream 22 into an elongated central transport stream 24' such that all cells in the central transport stream 24 can be responsive one by one. The sequence flows through the detection area 1042 of the microfluidic channel 1〇4. With this first-class design, it not only facilitates the subsequent cell sorting process, but also avoids the phenomenon that the microfluidic channel 104 is blocked due to the accumulation of cells. In addition, the detection device 12 will be described in detail. The detecting device I] is used to determine a particle that passes through the detecting region 1042 of the microfluidic channel 104 = No, a target particle. In this embodiment, since the microfluidic particle sorting wafer system 1 is a fluorescent excitation cell sorter, the detecting device 12 can determine whether the cells passing through the detecting area 1〇42 are detected by fluorescence detection. The cells have been fired by the cursor.
為了能具備偵測螢光之功能，偵測裝置Π可包含—於麵 及—判斷單元124。發射單元122可以發射-i光 f:tf通過偵測區域綱2之細胞。在實際應用中，ϊ、、則 ίϊ可以是—光學顯微鏡’而發射單元122則可以是J = f源’並可透過適當喊倾產生螢絲發光線。I 200940987 昭射判斷該細胞於該螢光激發光線之 長:螢光以利螢光之‘= 下，通=光線之照射 斷124判細物':=確ί 細胞並不是目標細胞。 Μ #1疋5亥In order to have the function of detecting fluorescence, the detecting device Π can include a face-and-judging unit 124. The transmitting unit 122 can emit -i light f:tf through the cells of the detection area. In practical applications, ϊ, 则 can be an optical microscope and the firing unit 122 can be a J = f source' and can generate a filament of light by appropriate shunting. I 200940987 Zhao Shot judges the length of the fluorescence of the cell in the fluorescence: fluorescing to the fluorescent ‘= under, pass=light illuminates 124 to judge the fines':= ί ί cells are not the target cells. Μ #1疋5海
子之=r若發的機制進行微粒 、要3 ^粒子為磁珠之類的磁性物質，# SL 4;?由磁糊之方式判斷通過_區域綱2 過雜較的微粒子。到貞測裝置12之 flj、.，口果為疋，代表通過_區域廟2之微粒子即為目 粒子，控制裝置14將會控制電極組1〇2產生.一力=目 標微粒子移動至目標區域綱4。 _力以將目 ，接下來，將就控制裝置14進行詳細之介紹。控 偵，裝置12及電極組1〇2。若伯測裝置12、之 判断'纟口果為疋，亦即通過偵測區域1〇 f的=裝置14將會控制電極組102產生一作用力^ ^ j移動至目標區域1044。也就是說，當細胞分選之程序完 系統1所分選出的目標細胞均會被收 在此實施例中，控制裝置14控制電極組〗〇2所產生之該 作，力為—介電泳力。所謂的介電泳_eetn)PhOTesis)是指^ 子文到不均自電場仙喊生移動之現象。當粒子於不 電場中受到極化時，由於受到不對稱之電場吸引力，因而粒 10 200940987 廠強或弱之方向移動。當介電泳力被用以操控粒 Τ須對粒子進行複雜的處理且不會對粒子造1 子。'c ^a此外，介電泳力可操控任何帶電或不帶電之粒 因，，^電泳技術近年來被廣泛地運用 ί用==、、細Γ蛋白f、DNA或奈米碳”。 ^ 迅’水力進行微粒子操控具有下列之優點：電訊梦切旅 和間短、晶片製程簡單、成本低以及不會影響細胞活性二、 ❹ e 在微饥體粒子分選晶n统丨巾，為了能提彳带 目標微粒子之移動，實際上，電極組1〇2 ;以一 =或《組形狀不對稱之第一電極及第二電極。舉例而今 ，二(Β)所示’第一電極1〇22及第二電極搬何^別^ 電極及長條狀電極，並且具有不同之電阻值。 ’、、、y 如圖二(A)所示，微流通道1〇4可 出口。於此實補中，兩個出口之寬度分別 300微米。作此一不對稱設計之目 」巧卡及 泳力作用之情況下，會固定由寬度較大的廢液田出I口在 電 而寬度較小的出口即為上述之目標區域1〇44。 /;u•出， 產生Ϊ = 步包含訊號 測區域觀之該細胞為目標細胞’亦即通過偵 號產生裝置I6產生-驅動訊號並傳送至^: 將會控制訊 102即根據該驅動訊號產生該作用力。 、、且102。電極組 在具際應用中，右藉由微粒子分連曰μ / 目標細胞包含不只-種罕見細胞 ^片糸統1所得到的 成U式者可利用不同的分 11 200940987 系Ί如磁性、電彳线榮光，繼續透過微粒子分選晶片 久插⑺目進彳次的細胞分選程序’即能將目標細胞所包含之 各種干見細胞分別篩選出來。 序之Ϊ外、’树過概錄子分選晶以、_行_分選程 ,,,必須先進行一些關於細胞樣本及晶片的準備工作， 序完成德、#將細巧樣本注人晶片等步驟。而#細胞分選程 5 還可繼續於該分選晶片上進行篩選後細胞之ΪΙ 率與一般細胞培養存活率近似。〜日^養所传之細胞存活 根據 片丰轉明之另—具體實施例為—種微流體粒子分選晶 目標 該微粒子分選晶片系統包含整及 區ΪΓ通運之—晶片1微流通道包含—偵測區域i 請參照圖 ❹ 所示，首先，該方作方法之流程圖。如圖三 ，測區域之一微粒子是ς為’該之 子可以是螢光標定微粒子、磁“°該目標微粒 :定：實崎’微粒子為細胞，而目標 位於 _ 擠成一中央輸送流，以係p认：3涊微粒子之樣 依序通過該伽m域。τ ;母個錄子均能—個接 之樣本流夹 個地 9 200940987 ^可包含多^子序之進行’上述之步驟 驟S112，發射—箸 所不，該方法可先執行子步 子。接著，該方法執J子:^4通過該細域之該微粒 激發光線之照射下是否^二，判斷該微粒子於該螢光 驟sm之兩種可能的判^結果^探^來，將分別就子步 弟種可能的判斷纟士果兔.》:止 是’亦即該微粒子為螢f ^ 之雜^ 子被判定為目_粒子時偵測區域鄉 ㈣，判定錄粒子不彳子，職方法執行步驟 作用力之影響子將；= ❹ 極组2!:::或ΐ,:力為-介電泳力。實際上，該電 以提補稱之第-電極及第二電極 電極之流式細胞儀，由觸^ 晶片中，並且針對不同的細胞樣 可避之選晶片系統 =月之微流體粒子分選晶片系統不i使用:: 之設備且不必外加高電壓，即可達到高純度細胞分選 13 200940987 篩選上， 述本發明&例之坪述’係希 施例來對本二以而並f以上述所揭露的較佳具體實 涵蓋各種改^目Γ:加以限制。相反地，其目的是希望能 具相等性的安排於本發明所欲中請之專利範 較佳具體實施例之詳述，姆望能更加清楚描 .揭 (? ❹ 明r:r月所申請：二的卿根 的轉，⑽越轉财可能的改 〇 14 200940987 【圖式簡單說明】 圖-鱗示根據本發明之第—具體實施綱微粒 3曰片糸統之功能方塊圖。 77、 之微流通道的示意圖。 圖二(B)係繪示整合於晶片上之電極組的一範例。If the mechanism of sub = r is to carry out the particle, the magnetic substance such as 3 ^ particle is magnetic bead, # SL 4;? It is judged by the method of magnetic paste to pass the microparticle of _ region class 2. The flj,., to the speculative device 12 is 疋, and the microparticles passing through the _region temple 2 are the target particles, and the control device 14 will control the electrode group 1 〇 2 to generate. One force = the target microparticle moves to the target region Outline 4. In the following, the control device 14 will be described in detail. Control, device 12 and electrode group 1〇2. If the device 12 determines that the mouthpiece is 疋, that is, the device 14 through the detection area 1 〇 f will control the electrode group 102 to generate a force ^ ^ j to move to the target area 1044. That is to say, when the cell sorting process is completed, the target cells sorted by the system 1 are received in this embodiment, and the control device 14 controls the electrode group to generate the action, the force is - dielectrophoretic force . The so-called dielectrophoresis _eetn) PhOTesis refers to the phenomenon that the sub-text is unevenly moved from the electric field. When a particle is polarized in a non-electric field, it is moved in a strong or weak direction due to the attractive electric field attraction. When dielectrophoretic forces are used to manipulate the particles, the particles are subjected to complex processing and do not create particles. 'c ^a In addition, the dielectrophoretic force can control any charged or uncharged particle, ^ electrophoresis technology has been widely used in recent years ί ==,, fine Γ protein f, DNA or nano carbon". 'Hydraulic microparticle manipulation has the following advantages: telecommunications dream travel and short, wafer process is simple, low cost and does not affect cell activity. ❹ e in micro-hungry particle sorting crystal n-type wipes, in order to The movement of the target microparticles, in fact, the electrode group 1〇2; with a = or "the first electrode and the second electrode with asymmetrical shape. For example, now, the second electrode 1" And the second electrode moves the electrode and the long electrode, and has different resistance values. ',, y As shown in Fig. 2 (A), the micro flow channel 1 〇 4 can be exported. In the case where the width of the two outlets is 300 micrometers respectively, in the case of the purpose of this asymmetric design, the effect of the swimming card and the swimming force will be fixed. That is, the target area 1〇44 described above. /;u•出, generate Ϊ = step includes the signal measurement area of the cell as the target cell', that is, generated by the Detector generation device I6-drive signal and transmitted to ^: The control signal 102 is generated according to the drive signal This force. , and 102. In the inter-application, the right-handed microparticles are connected to the 曰μ / target cells containing not only a rare cell, and the U-forms obtained by the U-type can be used for different points. 200940987 System, such as magnetic, electric彳 荣 荣 , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , In addition to the sequence, the tree has been selected for the cell sample and the wafer, and the preparation of the cell sample and the wafer must be performed first. step. The # cell sorting process 5 can also continue to screen on the sorting wafer and the cell ΪΙ rate is similar to the general cell culture survival rate. ～ ^ 养 养 养 养 细胞 细胞 细胞 养 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞Detection area i Please refer to Figure ,. First, the method is a flowchart of the method. As shown in Fig. 3, one of the particles in the measurement area is ' as 'the child can be a cursor with a cursor, a magnetic particle. The target particle: Ding: Shisaki' is a particle, and the target is located in a central transport stream. p recognizes: 3 涊 microparticles pass through the gamma domain τ; τ ; parental records can be connected to a sample stream 9 200940987 ^ can contain multiple ^ sub-orders of the above steps S112 , the method of transmitting - 箸 , 该 该 该 该 该 该 该 该 该 该 该 该 该 该 该 该 该 该 该 该 该 该 该 该 该 该 该 该 该 该 该 该 该 该 该 该 该 该 ^ ^ ^ ^ ^ ^ ^ ^ ^ ^ The two possible judgments ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ When the particle is detected in the area of the township (four), it is judged that the particle is not scorpion, and the influence of the force of the step method is performed; = ❹ pole group 2!::: or ΐ,: force is - dielectrophoretic force. Actually, the Electrolysing a flow cytometer called a first electrode and a second electrode electrode, by touching the wafer, and The same cell sample can avoid the selection of the wafer system = month microfluidic particle sorting wafer system does not use:: the device does not need to apply high voltage, can achieve high purity cell sorting 13 200940987 screening, the invention & The example of a syllabus is intended to cover the various aspects of the present disclosure and to limit the various modifications: instead, the purpose is to have an equivalence arrangement. In the detailed description of the preferred embodiment of the patent application of the present invention, Mwang can more clearly describe the disclosure (? ❹ r r: r application: two Qing's turn, (10) more money possible改〇14 200940987 [Simplified illustration of the drawings] Figure-Scale shows a functional block diagram of the microparticle 3 根据 根据 according to the first embodiment of the present invention. 77. Schematic diagram of the microfluidic channel. An example of an electrode set integrated on a wafer.
圖三係繪示根據本發明之第二具體實施例的微粒子 晶片糸統操作方法之流程圖。 、 圖四係繪示圖彡所不之步驟S11所包含的子步驟。 【主要元件符號說明】 SU〜S118 :流程梦驟 1 :微粒子分選晶片系統 10-·晶片 12 ·彳貞測模|且 14 :控制模組 102 ·電極組 104 :微流通道 1022 .苐一電極 1024 :第二電極 1042 :偵測區域 1044 :目標區域 122 :發射單元 124 :判斷單元 20 :邊鞘流 22 ·樣本流 24 :中央輪送流 26 I廢液出口 15Figure 3 is a flow chart showing a method of operating a microparticle wafer system in accordance with a second embodiment of the present invention. Figure 4 shows the sub-steps included in step S11. [Description of main component symbols] SU~S118: Process Dream 1: Microparticle sorting wafer system 10--Wafer 12 彳贞 模 模 | | 14: Control module 102 · Electrode group 104: Microfluidic channel 1022 Electrode 1024: second electrode 1042: detection area 1044: target area 122: emission unit 124: judgment unit 20: side sheath flow 22 • sample flow 24: central wheel flow 26 I waste liquid outlet 15