TW201105969A - Microfluidic device - Google Patents

Microfluidic device Download PDF

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TW201105969A
TW201105969A TW99122308A TW99122308A TW201105969A TW 201105969 A TW201105969 A TW 201105969A TW 99122308 A TW99122308 A TW 99122308A TW 99122308 A TW99122308 A TW 99122308A TW 201105969 A TW201105969 A TW 201105969A
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Taiwan
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substrate
flow channel
cells
microfluidic
light
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TW99122308A
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Chinese (zh)
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Gary P Durack
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Sony Corp
Sony Corp America
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  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)

Abstract

The present disclosure relates to microfluidic devices adapted for facilitating cytometry analysis of particles flowing therethrough. In certain embodiments, the microfluidic devices allow light collection from multiple directions. In certain other embodiments, the microfluidic devices use spatial intensity modulation. In certain other embodiments, the microfluidic devices have magnetic field separators. In certain other embodiments, the microfluidic devices have the ability to stack. In certain other embodiments, the microfluidic devices have 3-D hydrodynamic focusing to align sperm cells. In certain other embodiments, the microfluidic devices have acoustic energy couplers. In certain other embodiments, the microfluidic devices have phase variation producing lenses. In certain other embodiments, the microfluidic devices have transmissive and reflective lenses. In certain other embodiments, the microfluidic devices have integrally-formed optics. In certain other embodiments, the microfluidic devices have non-integral geographically selective reagent delivery structures. In certain other embodiments, the microfluidic devices have optical waveguides incorporated into their flow channels. In certain other embodiments, the microfluidic devices have optical waveguides with reflective surfaces incorporated into their flow channels. In certain other embodiments, the microfluidic devices have virus detecting and sorting capabilities. In certain other embodiments, the microfluidic devices display a color change to indicate use or a result.

Description

201105969 六、發明說明： 【發明所屬之技術領域】 本發明係關於微流體細胞測量系統。 【先前技術】 基於流動式細胞測量術的細胞分選技術，在2〇多年前首次 被引入研究社群中。其為一種已廣泛應用於生命科學研究之許 多領域的技術，充當諸如遺傳學、免疫學、分子生物學、及環 士兄科學專領域研究者的關鍵工具。不同於諸如免疫淘選或磁性 管柱分離類的整體細胞分離技術（bulk cdl sepamti〇n technique)，基於流動式細胞測量術的細胞分選儀，係以每秒 數千個、’’田胞或更南的速率連續地測量、分類、且接著分選個別 的細胞或粒子。此種針對單—細胞之快速「逐一」處理，使流 動式細胞測量術成為用以其他異質細胞懸浮液中萃取高純度 細胞亞群的獨特且有價值之工具。 以分選為目標的細胞通常用螢光物質以某種方式來標記。當 細胞通過緊密聚f、的高強度光束(典型地為雷射光束)時，結合 於細胞㈣光騎便發射”光^腦記錄下每—細胞之發射 強度。此料料接著Μ將每—細齡類，㈣於歡的分選 操作。基於流動式細胞測量術之細胞分選技術，已被成功應用 於數百種細胞類型、細胞成分、及微生物 具有可比較尺寸的無機粒子。 以及，許多類型之 流動式細胞㈣❹歧地應麟快速分㈣質細胞懸泮 099122308 4 201105969 液’以鑑別出成分亞群。使用流動式細胞測量細胞分選技術的 5午多應用之例子包括：分離稀少的用於aids研究的免疫系統 細胞群體、分離用於癌症研究的遺傳非典型細胞、分離用於遺 傳研究的特定染色體、及分離用於環境研究的不同種微生物。 舉例而言’營光標記的單株抗體經常用作鑑別諸如T淋巴細胞 及B淋巴細胞之類的免疫細胞的「標記物」，臨床實驗室通常 使用此技術來計算感染HIV之患者體内的「CD4陽性」T細 胞數目’而且，其亦使用此技術以鑑別與多種白血病及淋巴癌 相關聯的細胞。 近來，兩個受關注的領域正促使細胞分選技術轉向臨床、患 者護理應用，而非狹窄的研究應用。首先，為自化學醫藥開發 轉向生物醫藥開發。舉_言’許多新式癌症療法係基於^ 物。此等療法包括基於抗體的癌症治療劑之類型。基於細胞測 量的細胞分選儀，可在此等產品之鑑別、開發、純化、及最終 製造中起重要作用。 與此相關的是轉向使用細胞替代療法進行患者護理。當前對 幹細胞之關注大部分係圍繞通常稱作再生療法或再生醫學的 醫學新領域。此等療法可經常需要自患者組織分離大量之相對 稀少的細胞。舉例而言，成體幹細胞可自骨髓分離出來，且最 終用作為再輸注物之一部分，再返回至其所移出的患者中。= 胞測量術本身極適用於此等療法。 、 滴式細 現今廣泛使用的細胞分選儀有兩種基本類型。其為 099122308 5 201105969 胞分選儀(droplet cell sorter)」及「流體切換式細胞分選儀(fluid switching cell sorter)」。滴式細胞分選儀利用微液滴作為容器， 將所選細胞輸送至收集容器中。微液滴係藉由將超音波能量柄 聯成喷射射流所形成。接著，含有被選擇用於分選的細胞的液 滴’被靜電方式導引至預定位置。此為極有效之方法，每秒可 自單液流中分選出多達90,000個細胞，其侷限之處主要在於， 液滴產生頻率及照射所需時間。 先前技術之流動式細胞測量系統詳細描繪於Durack等人之 美國已公開專利申請案第US 2005/0112541 A1號中。 然而，滴式細胞分選儀並不特別具有生物安全性。作為液滴 形成過程之-部分而產生的氣_，可帶有對生物有害的物 質。為此’生物安全性滴式細胞分選儀得以開發，其係含钟在 生物安全箱内，以使其可在基本上為封閉的環境内操作。令人 遺慽的是，此_之系統並不適合於在臨床環境下對患者樣品 進行常規分選所需之無菌狀態及操作者保護。 第二種類型之基於流動式細胞測量之細胞分選儀為流體切 換式細胞分選儀。大錄越切換式細胞分選儀细壓電裝置 系統’將一段流動樣品流導入收集容器中。與梅 胞为遥儀相比，流體切換式細胞分選儀由於用於導汽樣”的 機械系狀獅，㈣有更_最大細齡解=週=、 始樣品導流與穩定未分選流紐時之_日蝴通常明顯大: 滴式細胞分縣上之_產生__。崎長軸使流體切 099122308 6 201105969 換式細胞分選儀限制於每秒數百個細胞之處理速率。出於相同 原因，由流體細胞分選儀所切換的液流段的體積，通常為來自 液滴產生器的單-微液滴的至少1G倍^此料致流體切換式 釦選儀之收集容器中的細胞濃度相應地低於滴式分選儀之收 集容器。 新一代微流體技術，為了提高流體切換裝置之效率、及提供 在概念上類似於電子積體電路的晶片上之細胞分選能力，提供 了極大希望。許多微流體系統已表明可成功地自異質細胞群體 分選細胞。其具有以下優點：完全獨立、易於滅菌、且可在按 拋棄式零件考慮以足夠規模(利用所得製造效率)製造。 原型的微流體裝置例示於圖丨中，且整體上以編號1〇表示。 微流體裳置ίο包含基板12，其具有藉由本技藝中已知的任何 適宜方法形成於其中的流體流動通道14。基板12可由玻璃、 塑膠、或任何其他適宜的材料形成，且實質上可呈透明、或其 一部分實質上呈透明。基板12另外具有耦接至其中的三個埠 16 18及20。蜂16為顆液(sheath fluid)之入口皡。槔16具有 與流體流動通道22流體連通的中心轴向通路，而該流體流動 通道22係連接流體流動通道14，以使自外部供應源(未圖示） 進入埠16的鞘液進入流體流動通道22，且接著流入流體流動 通道14。鞘液供應源可藉由熟悉本技藝者已知的任何適宜耦 接機構連接至埠16。 埠18亦具有中心軸向通路’其係經由樣品注入管24而與流 099122308 201105969 體流動通道Μ呈流體連^樣品注人f 24被定位成與流體流 動通道Μ之縱向軸同轴。因此，將液態細胞樣品注入璋18中， 而同時將鞘液^主入埠16中，將會導致細胞流過被勒液包圍的 抓體肌動通道14。流體流動通道14和22及樣品注人管以之 尺寸及、.减被選擇成為使得鞘液/樣體在通過裝置1〇時 將展現層w ’如本技藝巾已知。埠2()搞接至流體流動通道Μ 之末端’以使難/樣品流體可自微流體裝置ι〇移出。 當勒液/樣品流體流過流體流動通道14時，可使用細胞測量 技術，藉由使照射源照射通過基板12且進入流體流動通道Μ 中介於樣品注入管24與出口埠2〇之間的某一點，而加以分 析。另外，微流體裳置10可加以修改，以提供細胞分選操作， 如本技藝中已知。 儘管與上文所述_之基本㈣料置已被證明工作良 好’但先前技射仍需要制賴流體裝置之細朗量系統^ 行改良。本發明著力於滿足此需要。 【發明内容】 本發明係關於適合於對流經的粒子便利地進行細胞測量分 析的微流體裝置。在某些具體射，該等微流體裝置允許自多 個方向收集光。在某些其他具體财，微流體裝置使用空間強 度調變。在某祕料體射，微㈣心具有磁場分離哭。 在某些其他具體例中’微流體裝置具有堆叠能力。在某些盆他 具體例中，微流财置具有3①流體動力學聚焦來對齊精细 099122308 8 201105969 胞。在某些其他具體例中’微流體裝置具有音能搞合器。在某 些其他具體例中’微流體裝置具有產生相位變化的透鏡。在某 些其他異體例中’微流體裝置具有透射鏡及反射鏡。在某些其 他具體例中’微流體裝置具有整體式形成的光學器件。在禁些 其他具體例中，微流體裝置具有非整體式區塊選擇性試劍傳遞 結構。在某些其他具體例中，微流體裝置具有併入其流動通道 中的光波導器。在某些其他具體例中’微流體裝置具有併入其 流動通遒中而連同有反射面的光波導器。在某些其他具體例 中，微流體襄置具有病毒偵測及分選能力。在某些其他具體例 中’微流體裝置顯示出顏色改變以指示其使用或結果。 在一具體例中，揭示一種微流體裝置系統，包含：基板；形 成於該基板中的微流體流動通道，其中，該流動通道延伸通過 該基板之一部分’此部分適合於對流入該流動通道的細胞便利 地進行細胞測量分析；可用於在第-方向上收集由該等細胞發 射的光的第一光收集裝置，該第一光收集裝置產生第一輸出； 可用於在第二方向上收集由該等細胞發射的光的第二光收集 • 織’該第二光收鍊置產生第二輸出；及，可驗狐該^ 一輸出及該第二輸出的偵測光學器件。 在另一具體例中，揭示一種偵測樣品中粒子的方法，此方法 包含以下步驟：a)設置-個微流體裝置，該微流體裝置包含： 基板、及形成於該基板中的微流體流動通道，其中，哼流動通 道延伸通過該基板之-部分，此部分適合於對流入該流動通道 099122308 9 201105969 的細胞便利地進行細胞測量分析’ b)在第一方向上捕獲自該等 細胞發射的第一光；c)在第·一方向上捕獲自5亥專細胞發射的第 二光；d)組合在步驟⑻及⑹捕獲的該第一光與該第二光；及 e)對該組合的第一光與第二光進行細胞測量分析。 在另一具體例中，揭示一種偵測樣品中粒子的方法，此方法 包含以下步驟：a)設置一個微流體裝置’該微流體裝置包含： 基板；形成於該基板中的第一微流體流動通道，其中，該第一 流動通道延伸通過該基板之一部分’此部分適合於對流入該第 一流動通道的第一細胞便利地進行細胞測量分析；及，形成於 該基板中的第二微流體流動通道，其中，該第二流動通道延伸 通過§亥基板之一部分，此部分適合於對流入該第二流動通道的 第二細胞便利地進行細胞測量分析；b)產生瞄準於該第一流動 通道及s亥第·一流動通道的激發光束，c)在該激發光束到達該第 一流動通道之前，以第一方式在空間上改變該激發光束；及， d)在該激發光束到達該第二流動通道之前，以第二方式在空間 上改變該激發光束。 在又一具體例中，揭示一種微流體裝置，包含：基板；形成 於該基板中的微流體流動通道，其中，該流動通道延伸通過該 基板之一部分，此部分適合於對流入該流動通道的細胞便利地 進行細胞測量分析；形成於該基板板上(〇nb〇ard)的樣品收容 孔，該樣品收容孔係流體式耦接至該流動通道；及，裝設於該 基板板上、且當被激發時可用於在該樣品收容孔内產生磁場的 099122308 10 201105969 電磁體。 在又一具體例中’揭示-種微流體裝置，包含:基板;形成 於該基板中的微流體流動通道，其中，該流動通道延伸通過該 基板之-部分，此部分適合於職人誠動通道的細胞便利地 • 進行細胞測量分析；及，至少—條位於該基板之表面上的腿 (1 e g)’該至少一條腿有助於將該微流體裝置堆疊於另一微流體 裝置上。 在另一具體例中，揭示一種微流體裝置，包含：基板；形成 於該基板中的微流體流動通道，其中，該流動通道延伸通過該 基板之一部分，此部分適合於對流入該流動通道的細胞便利地 進行細胞測量分析；及，至少一個流體式耦接至該流動通道的 流體動力學對齊結構，該至少一個流體動力學對齊結構可用於 定向該等細胞，使得，大部分該等細胞據其最大尺寸來分析。 在又一具體例中，揭示一種微流體裝置，包含：基板；形成 於該基板中的微流體流動通道，其中，該流動通道延伸通過該 基板之一部分’此部分適合於對流入該流動通道的細胞便利地 進行細胞測量分析；裴設於該基板板上的孔，該孔與該流動通 道流體式耦接；及，裝設於該孔内的音能耦合器。 在又一具體例中，揭示一種微流體裝置，包含：基板；形成 於该基板中的微流體流動通道，其中，該流動通道延伸通過該 基板之一部分，此部分適合於對流入該流動通道的細胞便利地 進行細胞測量分析；及，形成於該基板板上的透鏡，該透鏡有 099122308 11 201105969 效於在空間上改變通過其間的光之強度。 在另一具體例中’揭示—種微流體裝置，包含：基板；形成 於該基板中的微流體流動通道，其中，該流動通道延伸通過該 基板之σ卩分，此部分適合於對流入該流動通道的細胞便利地 進行、’’田胞測里分析，形成於該基板板上的第一透鏡，該第一透 鏡裝設於該流動通道之第-側；及，形成於板板上的第二 透鏡’該第二透鏡裝設於該流動通道之第二側。 在又一具體例中，揭示一種微流體裝置，包含：具有第一表 面的基板；形成於該基板中的微流體流動通道，其中，該流動 通道延伸通過該基板之一部分，此部分適合於對流入該流動通 道的細胞便利地進行細胞測量分析；及’在該第一表面形成於 該基板板上的透鏡，該透鏡處於該第一表面下方。 在又一具體例中，揭示一種微流體裝置，包含：具有第一表 面的基板；形成於該基板板上的試劑收容孔；具有裝設於其上 方的試劑的試劑結構’其中，將該試劑結構施加於該第一表 面，使該試劑與該試劑收容孔對齊，以便轉移試劑至其中；及， 形成於該基板中的微流體流動通道，其中，該流動通道係流體 式耦接至該試劑收容孔。 在又一具體例中’揭示一種微流體裝置，包含：基板；形成 於該基板中的微流體流動通道，其中，該流動通道延伸通過該 基板之一部分’此部分適合於對流入該流動通道的細胞便利地 進行細胞測量分析；及’形成於該流動通道中的光波導器。 099122308 12 201105969 在另一具體例中’揭示一種微流體裝置，包含：基板；形成 於該基板中的微流體流動通道’其中，該流動通道延伸通過該 基板之一部分，此部分適合於對流入該流動通道的細胞便利地 進行細胞測量分析；形成於該流動通道中的光波導器；及，裝 設於該流動通道内的反射面。 在又一具體例中，揭示一種測定藥理學功效的方法，此方法 包含以下步驟：a)設置一個微流體裝置，該微流體裝置包含： 基板；形成於該基板中的微流體流動通道，其中，該流動通道 延伸通過該基板之一部分，此部分適合於對流入該流動通道的 病毒粒子便利地進行細胞測量分析；及，形成於該基板板上的 孔；b)將一物質沉積於該孔中；c)使該等病毒粒子流入該孔中； d)使該等病毒粒子與該物質反應；及，e)基於該反應以測定該 物質之藥理學功效。 在又一具體例中，揭示一種偵測樣品中粒子的方法，此方法 包含以下步驟：a)設置一個微流體裝置，該微流體裝置包含： 基板；形成於該基板中的微流體流動通道，其中，該流動通道 延伸通過該基板之一部分，此部分適合於對流入該流動通道的 細胞便利地進行細胞測量分析；及，形成於該基板板上的染料 貯存器(dye repository); b)將一染料沉積於該孔中；c)對該等細 胞進行細胞測量分析；及，d)在該細胞測量分析完成之後，使 該染料離開該染料貯存器，且進入該流動通道。 亦揭示其他具體例。 099122308 13 201105969 【實施方式】 出於促進理解本發明原理之目的，現將參考圖式中例示之具 體例’且專用語言將用以描綠該等具體例。然而，應瞭解1 不意欲藉此限制本發明之範蜂，所例示的裝置之此等變化及進 -步修改’及如此處所齡的本糾之原理之料進一步應 用’預期通常將為熟悉本發明相關技藝者基於本案申請專利範 圍所想到。 自多個方向收集光的微流體裝置： 本發明之某些具體例大體而言係關於在微流體裝置上使用 細胞測量(諸如’流動式細胞測量術或影像細朗量術)來分析 樣品的系統。為在細胞測量操作期間制或鐘別細胞，電磁輕 射(諸如，可見光)來源(諸如，雷射器）被引導至偵測區。當細 胞通過偵測區時’光源使細胞發螢光。此效應可藉由在細胞到 達谓測區之前添加的某些染料來增強。在大多數系統中，光源 僅施加於細胞測量裝置之—側，偵測光學器件亦僅债測來自樣 品細胞-側之榮光。如此限制了自營光樣品細胞捕獲的光子之 數因為’自其他方向發射的光子並不被侧光學器件所捕 獲。光強度可被提rfj而產生更多以螢光方式產生的光子，然 而，如此亦增加了信號中雜訊之量。 為儘可能多地捕獲由發榮光的樣品細胞所產生的光子，光源 可施加於侧區之多側。另外，傾測光學器件亦可置於價測區 之多側。如此使得該系統對於既定的激發光強度可捕獲更多的 099122308 14 201105969 發射光子， 而不增加接收信號中之雜訊 。此外，201105969 VI. Description of the Invention: TECHNICAL FIELD OF THE INVENTION The present invention relates to a microfluidic cell measurement system. [Prior Art] The cell sorting technique based on flow cytometry was first introduced into the research community more than 2 years ago. It is a technology that has been widely used in many fields of life science research and serves as a key tool for researchers in the fields of genetics, immunology, molecular biology, and neuroscience. Unlike bulk cdl sepamti〇n techniques such as immunopanning or magnetic column separation, cell sorters based on flow cytometry are thousands of cells per second. The rate at or south is continuously measured, sorted, and then sorted into individual cells or particles. This rapid “one-by-one” treatment of single-cells makes flow cytometry a unique and valuable tool for extracting high-purity cell subpopulations from other heterogeneous cell suspensions. Cells targeted for sorting are usually labeled with a fluorescent substance in some way. When the cells pass through a high-intensity beam (typically a laser beam) that is tightly condensed, the cells are combined with the cell (4) to ride the light. The light intensity of each cell is recorded. This material is then 每Fine-aged, (d) Yuhuan sorting operation. Cell sorting technology based on flow cytometry has been successfully applied to hundreds of cell types, cell components, and inorganic particles of comparable size. Many types of flow cells (4) ❹ 地 地 应 快速 快速 快速 快速 四 四 四 四 四 四 四 四 四 99 99 99 99 99 99 99 99 99 99 99 99 99 99 99 99 99 99 99 99 99 99 99 99 99 99 99 99 99 99 99 99 99 99 99 99 99 99 99 99 99 Immune system cell populations for aids research, isolation of genetic atypical cells for cancer research, isolation of specific chromosomes for genetic research, and isolation of different species of microorganisms for environmental research. For example, 'camera-marked Individual antibodies are often used as "markers" for identifying immune cells such as T lymphocytes and B lymphocytes, which are commonly used in clinical laboratories. Technique that counts the number of HIV infected patients is the "CD4 positive" T cells "Furthermore, it is also the use of this technique in more differentiated cell lymphoma and leukemia associated. Recently, two areas of concern are driving cell sorting technology to clinical, patient care applications rather than narrow research applications. First, we are shifting from chemical medicine development to biomedical development. Many new cancer therapies are based on thyrus. Such therapies include types of antibody-based cancer therapeutics. Cell sorting based cell sorters play an important role in the identification, development, purification, and final manufacture of these products. Related to this is the shift to patient care using cell replacement therapy. Much of the current focus on stem cells is centered around new areas of medicine commonly referred to as regenerative or regenerative medicine. These therapies can often require the isolation of a large number of relatively rare cells from the patient's tissue. For example, adult stem cells can be isolated from the bone marrow and eventually used as part of a reinfusion and returned to the patient from which they were removed. = Cell measurement itself is ideal for these therapies. Drip type There are two basic types of cell sorters that are widely used today. It is 099122308 5 201105969 Droplet cell sorter and "fluid switching cell sorter". The drop cell sorter uses the microdroplets as a container to deliver the selected cells to a collection container. The microdroplets are formed by joining the ultrasonic energy handles into jet jets. Next, the droplets containing the cells selected for sorting are electrostatically guided to a predetermined position. This is an extremely efficient method for sorting up to 90,000 cells per second from a single stream, with limitations mainly in the frequency of droplet generation and the time required for irradiation. A flow cytometry system of the prior art is described in detail in U.S. Published Patent Application No. US 2005/0112541 A1 to the name of the entire disclosure of the entire disclosure. However, drop cell sorters are not particularly biosafe. The gas produced as part of the droplet formation process may carry substances harmful to living things. To this end, a biosafety drip cell sorter has been developed which incorporates a clock in a biosafety box to allow it to operate in a substantially enclosed environment. It is awesome that this system is not suitable for the sterility and operator protection required for routine sorting of patient samples in a clinical setting. The second type of cell sorter based on flow cytometry is a fluid exchange cell sorter. The large-volume switching cell sorter fine piezoelectric device system 'puts a flow of sample stream into the collection container. Compared with the U.S. cell phone, the fluid-switched cell sorter has a more _maximum aging solution = week =, initial sample diversion and stable unsorted because of the mechanical lion-like lion used for the steam-guided sample. When the flow is _, the day of the butterfly is usually significantly larger: the drop cell is divided into ___ _. The long axis makes the fluid cut 099122308 6 201105969 The exchange cell sorter is limited to the processing rate of hundreds of cells per second. For the same reason, the volume of the liquid flow section switched by the fluid cell sorter is typically at least 1 G times that of the single-micro droplets from the droplet generator. The cell concentration in the drop is correspondingly lower than that of the drop sorter. A new generation of microfluidic technology, in order to improve the efficiency of the fluid switching device, and to provide cell sorting capability on a wafer that is conceptually similar to an electronic integrated circuit. There is great hope that many microfluidic systems have been shown to successfully sort cells from heterogeneous cell populations. They have the following advantages: they are completely self-contained, easy to sterilize, and can be considered at a sufficient scale in disposable parts (using the resulting manufacturing efficiency) The prototype microfluidic device is illustrated in Figure , and is generally indicated by the number 1 。. The microfluidic device comprises a substrate 12 having fluid flow formed therein by any suitable method known in the art. Channel 14. Substrate 12 may be formed of glass, plastic, or any other suitable material, and may be substantially transparent, or a portion thereof being substantially transparent. Substrate 12 additionally has three turns 16 18 and 20 coupled thereto. The bee 16 is the inlet port of the sheath fluid. The crucible 16 has a central axial passage in fluid communication with the fluid flow passage 22, and the fluid flow passage 22 is connected to the fluid flow passage 14 to provide an external supply source (not The sheath fluid entering the crucible 16 enters the fluid flow channel 22 and then flows into the fluid flow channel 14. The sheath fluid supply can be coupled to the crucible 16 by any suitable coupling mechanism known to those skilled in the art. Having a central axial passage 'through the sample injection tube 24 and fluidly connected to the flow 099122308 201105969 body flow passage ^ ^ sample injection f 24 is positioned with the fluid flow passage Μ longitudinal Coaxial. Therefore, injecting a liquid cell sample into the crucible 18 while simultaneously introducing the sheath fluid into the crucible 16 will cause the cells to flow through the grasping body muscle channel 14 surrounded by the fluid. Fluid flow channels 14 and 22 And the size of the sample injection tube is selected such that the sheath liquid/sample will exhibit a layer w' as it passes through the device 1 'as known in the art towel. 埠 2 () is engaged to the fluid flow path Μ The end 'to allow the difficult/sample fluid to be removed from the microfluidic device. When the liquid/sample fluid flows through the fluid flow channel 14, cell measurement techniques can be used by illuminating the substrate through the substrate 12 and into the fluid. The flow channel Μ is at a point between the sample injection tube 24 and the outlet 埠2〇 and is analyzed. Additionally, the microfluidic skirt 10 can be modified to provide a cell sorting operation, as is known in the art. Although the basic (4) material set described above has been proven to work well, the prior art still requires a fine-grained system to improve the fluid device. The present invention is focused on meeting this need. SUMMARY OF THE INVENTION The present invention is directed to a microfluidic device suitable for conveniently performing cell measurement analysis on particles flowing therethrough. In some specific shots, the microfluidic devices allow light to be collected from multiple directions. In some other specific financial applications, microfluidic devices use spatial intensity modulation. In a secret body shot, the micro (four) heart has a magnetic field to separate and cry. In some other specific examples, the microfluidic device has a stacking capability. In some specific examples, the microfluid has 31 fluid dynamics focus to align the fine 099122308 8 201105969 cells. In some other specific examples, the microfluidic device has a sound energy combiner. In some other specific examples, the microfluidic device has a lens that produces a phase change. In some other variants, the microfluidic device has a transmission mirror and a mirror. In some other embodiments, the microfluidic device has integrally formed optics. In other specific examples, the microfluidic device has a non-integral block selective test-sword transfer structure. In certain other embodiments, the microfluidic device has an optical waveguide incorporated into its flow channel. In some other specific examples, a microfluidic device has an optical waveguide incorporated into its flow wanted with a reflective surface. In some other specific examples, the microfluidic device has virus detection and sorting capabilities. In some other specific examples, the microfluidic device exhibits a color change to indicate its use or result. In one embodiment, a microfluidic device system is disclosed, comprising: a substrate; a microfluidic flow channel formed in the substrate, wherein the flow channel extends through a portion of the substrate 'this portion is adapted to flow into the flow channel The cells are conveniently subjected to cell measurement analysis; a first light collection device operable to collect light emitted by the cells in a first direction, the first light collection device generating a first output; The second light collection of the light emitted by the cells is woven by the second light-receiving chain to produce a second output; and the detection optical device of the second output and the second output. In another embodiment, a method of detecting particles in a sample is disclosed, the method comprising the steps of: a) providing a microfluidic device comprising: a substrate, and a microfluidic flow formed in the substrate a channel, wherein the 哼 flow channel extends through a portion of the substrate, the portion being adapted to facilitate cell measurement analysis of cells flowing into the flow channel 099122308 9 201105969' b) capturing from the cells in a first direction First light; c) capturing the second light emitted from the 5th cell in the first side; d) combining the first light and the second light captured in steps (8) and (6); and e) The first light and the second light are subjected to cell measurement analysis. In another embodiment, a method of detecting particles in a sample is disclosed, the method comprising the steps of: a) providing a microfluidic device comprising: a substrate; a first microfluidic flow formed in the substrate a channel, wherein the first flow channel extends through a portion of the substrate 'this portion is adapted to facilitate cell measurement analysis of a first cell flowing into the first flow channel; and a second microfluidic formed in the substrate a flow channel, wherein the second flow channel extends through a portion of the substrate, the portion being adapted to facilitate cell measurement analysis of the second cell flowing into the second flow channel; b) generating an aiming at the first flow channel And an excitation beam of the first flow channel, c) spatially changing the excitation beam in a first manner before the excitation beam reaches the first flow channel; and, d) reaching the second in the excitation beam The excitation beam is spatially changed in a second manner prior to the flow channel. In yet another embodiment, a microfluidic device is disclosed, comprising: a substrate; a microfluidic flow channel formed in the substrate, wherein the flow channel extends through a portion of the substrate, the portion being adapted to flow into the flow channel The cell is conveniently subjected to cell measurement analysis; a sample receiving hole formed on the substrate plate, the sample receiving hole is fluidly coupled to the flow channel; and is mounted on the substrate plate, and When excited, it can be used to generate a magnetic field in the sample receiving hole of 099122308 10 201105969 electromagnet. In another embodiment, a microfluidic device is disclosed, comprising: a substrate; a microfluidic flow channel formed in the substrate, wherein the flow channel extends through a portion of the substrate, the portion is suitable for a person's honesty channel The cells conveniently • perform a cell measurement analysis; and, at least — a leg (1 eg) on the surface of the substrate that facilitates stacking the microfluidic device on another microfluidic device. In another embodiment, a microfluidic device is disclosed, comprising: a substrate; a microfluidic flow channel formed in the substrate, wherein the flow channel extends through a portion of the substrate, the portion being adapted to flow into the flow channel The cells are conveniently subjected to cell measurement analysis; and at least one fluidically coupled to the fluid dynamic alignment structure of the flow channel, the at least one fluid dynamic alignment structure being operable to orient the cells such that a majority of the cells are Its maximum size is analyzed. In yet another embodiment, a microfluidic device is disclosed comprising: a substrate; a microfluidic flow channel formed in the substrate, wherein the flow channel extends through a portion of the substrate 'this portion is adapted to flow into the flow channel The cells are conveniently subjected to cell measurement analysis; a hole disposed on the substrate plate, the hole being fluidly coupled to the flow channel; and a sound energy coupler disposed in the hole. In yet another embodiment, a microfluidic device is disclosed, comprising: a substrate; a microfluidic flow channel formed in the substrate, wherein the flow channel extends through a portion of the substrate, the portion being adapted to flow into the flow channel The cells are conveniently subjected to cell measurement analysis; and a lens formed on the substrate plate having 099122308 11 201105969 effective in spatially varying the intensity of light passing therethrough. In another embodiment, a microfluidic device is disclosed, comprising: a substrate; a microfluidic flow channel formed in the substrate, wherein the flow channel extends through a σ of the substrate, the portion being adapted to flow into the The cells of the flow channel are conveniently subjected to ''field analysis, a first lens formed on the substrate plate, the first lens being mounted on the first side of the flow channel; and formed on the plate The second lens 'the second lens is mounted on the second side of the flow channel. In yet another embodiment, a microfluidic device is disclosed comprising: a substrate having a first surface; a microfluidic flow channel formed in the substrate, wherein the flow channel extends through a portion of the substrate, the portion being adapted to The cells flowing into the flow channel are conveniently subjected to cell measurement analysis; and a lens formed on the first surface of the substrate, the lens being below the first surface. In still another embodiment, a microfluidic device is disclosed, comprising: a substrate having a first surface; a reagent receiving hole formed on the substrate plate; and a reagent structure having a reagent disposed thereon, wherein the reagent a structure applied to the first surface to align the reagent with the reagent receiving aperture to transfer a reagent thereto; and a microfluidic flow channel formed in the substrate, wherein the flow channel is fluidly coupled to the reagent Containing holes. In another embodiment, a microfluidic device is disclosed comprising: a substrate; a microfluidic flow channel formed in the substrate, wherein the flow channel extends through a portion of the substrate 'this portion is adapted to flow into the flow channel The cells are conveniently subjected to cell measurement analysis; and 'optical waveguides formed in the flow channel. 099122308 12 201105969 In another embodiment, a microfluidic device is disclosed, comprising: a substrate; a microfluidic flow channel formed in the substrate, wherein the flow channel extends through a portion of the substrate, the portion being adapted to flow into the The cells of the flow channel are conveniently subjected to cell measurement analysis; an optical waveguide formed in the flow channel; and a reflective surface disposed in the flow channel. In yet another embodiment, a method of determining pharmacological efficacy is disclosed, the method comprising the steps of: a) providing a microfluidic device comprising: a substrate; a microfluidic flow channel formed in the substrate, wherein a flow channel extending through a portion of the substrate, the portion being adapted to facilitate cell measurement analysis of virions flowing into the flow channel; and a hole formed in the substrate plate; b) depositing a substance in the hole And c) causing the virions to flow into the pore; d) reacting the virions with the material; and, e) determining the pharmacological efficacy of the virion based on the reaction. In yet another embodiment, a method of detecting particles in a sample is disclosed, the method comprising the steps of: a) providing a microfluidic device comprising: a substrate; a microfluidic flow channel formed in the substrate, Wherein the flow channel extends through a portion of the substrate, the portion being adapted to facilitate cell measurement analysis of cells flowing into the flow channel; and a dye reservoir formed on the substrate plate; b) A dye is deposited in the well; c) a cell measurement analysis is performed on the cells; and, d) after the cell measurement analysis is completed, the dye exits the dye reservoir and enters the flow channel. Other specific examples are also disclosed. MODE FOR CARRYING OUT THE INVENTION For the purpose of promoting the understanding of the principles of the present invention, reference will now be made to However, it should be understood that 1 is not intended to limit the invention, and such variations and modifications of the illustrated device and the further application of the principles of the present invention as herein are intended to be familiar with Those skilled in the art will be aware of the scope of the patent application in this application. Microfluidic devices that collect light from multiple directions: Some specific examples of the invention generally relate to the use of cellular measurements (such as 'flow cytometry or image sizing') to analyze samples on a microfluidic device. system. To make or clock cells during cell measurement operations, electromagnetic light (such as visible light) sources (such as lasers) are directed to the detection zone. When the cells pass through the detection zone, the light source fluoresces the cells. This effect can be enhanced by the addition of certain dyes before the cells reach the pre-measurement zone. In most systems, the light source is applied only to the side of the cell measurement device, and the detection optics only pays for the glory from the cell side of the sample. This limits the number of photons captured by the self-operated light sample cells because photons emitted from other directions are not captured by the side optics. The light intensity can be increased by rfj to produce more photons that are produced in a fluorescent manner. However, this also increases the amount of noise in the signal. To capture as much of the photons produced by the glory sample cells, the source can be applied to multiple sides of the side regions. In addition, the tilting optics can also be placed on multiple sides of the price measurement zone. This allows the system to capture more of the 099122308 14 201105969 emitted photons for a given excitation light intensity without increasing the noise in the received signal. In addition,

θ不思性例示使用細胞測量來分析樣品的系 p之雜訊。此外’該方法將自兩 單一光偵測器的單一準直路徑 二伯測器之改良。使用兩個光偵 b雜訊加倍。因此，使用單一光 統200。系統 〇〇—可包3形成於基板2G2上的微流體裝置(在此處以側視圖 ‘、’、頁不）其具有含納於其巾的偵測流動通道2Q4。出於簡單及易 圖2顯示基板2〇2 β之單一通道。然而，應瞭解 單通道可為熟悉本技藝者可想到的多個細胞測量通道及多 種可此的通道組構的代表例。各種其他的細胞測量組件亦可含θ does not exemplify the use of cell measurements to analyze the noise of the sample p. In addition, the method will be improved from a single collimation path of the two single photodetectors. Use two light detection b noise to double. Therefore, a single optical system 200 is used. System 〇〇 - The microfluidic device (which may be in the side view ', ', no page) formed on the substrate 2G2 has a detection flow channel 2Q4 contained in the towel. For simplicity and ease Figure 2 shows a single channel of substrate 2〇2 β. However, it should be understood that a single channel can be a representative of a plurality of cell measurement channels and a variety of channel configurations that are within the skill of the art. Various other cell measurement components may also be included

系統200可另外包含激發光源2〇6、多芯光纖麟218、聚 焦透鏡212及220、及偵測光學器件226。透鏡212及22〇可 置於基板202之外侧’如圖2所例示，或可視特定應用之需要 及成本考慮之不同，安裝於或整體式形成於基板2〇2内。激發 光源206可包含雷射器或此技藝中已知的其他電磁輻射來 源。偵測光學器件226可包含此技藝中已知的各種感光手段 (light sensing means)，堵如，光電倍增管、光電二極體、或突 崩光電二極體。 在操作中，激發光源206(通常為雷射器)將光束引向二向色 鏡(dichroic mirror)208。該二向色鏡被組構成將自光源2〇6所 099122308 15 201105969 接收的波長反射至晶片加上之偵測區別。各種其他的光學 水焦組件(諸如’透鏡212)可用以確保激發光源適當聚焦 於備測區210上。當穿越微流體流動通道204的樣品細胞214 通過偵測區21〇且發出螢光時，所產生的光子在多個方向上發 射。-些光子將朝聚焦透鏡212反向發射，通過二向色鏡 2〇8(其被組構成讓螢光波長通過），且進入多芯光纖繞線叫 之第-末端2】6。自晶片2〇2之對側發射的光子，將選擇地被 透鏡220聚焦，且進入光纖纜線218之第二末端。所組合 的光子群接著將自光賴線218之第三末端224發射，且被= 測光學器件226接收。偵測光學器件226可接著使來自兩個光 纖芯(fiber optic core)之组合發射呈準直，且將該發射傳輸至— 或多個光偵測器（未圖示），以供測量。在某些具體例中，—系 歹J之一向色及帶通(bandpass)濾光器將用來選擇特定的波長 帶，且將其傳輸至光偵測器。在其他具體例中，不使用多芯光 纖纜線，分別的偵測光學器件可用來捕獲所發射的光子。 圖3示意性例示本發明之另一具體例，此處顯示系統3〇〇， 其中，多個光源306用來在樣品細胞314中產生螢光。在此具 體例中，自樣品細胞314反射及傳輸的光子均將自細胞測量晶 片302之兩側得到偵測。又，所發射的光子將進入光纖纜線 318之末端3丨6及322 ’且被偵測光學器件326所捕獲。在某 些具體例中，光源306可被組構成產生不同波長的光。偵測光 學器件可接著偵測已被染色而在不同波長下發螢光的不同類 099122308 16 201105969 型的細胞。 使用空間強度調變的微流體裝置： 本發明之某些具體例大體上而言係關於在微流體裝置上使 ^胞測量術(諸如，流動式細胞測量術或影像細胞測量術)來 刀析樣品的系統。為提高總細胞通量’多個流動通道可平行使 用，以自既定的樣品分選細胞。在此等系統中，偵測光學器件 關的電子處理設備必須區分自個別的通道所接收的信 唬。實現此舉之—種方式為在各通道上使用各自具有其自身波 長的個別激發光源。然而，如此為系統增加了大量成本及複雜 性。單一光源亦可以重複方式在通道間掃描，使偵測光學器件 基於時間而確定哪個通道被掃描。然而’由於有可能細胞通過 既疋通道之偵測區域，而光源激發了不同通道之偵測區域，所 以’此方法亦傾向於出錯。 如本文所揭示，區分個別的通道螢光信號之另一方式為將單 -激發光關料導至所有通道’但，在㈣上改變或成形激 發光束圖型(excitation beam pattern)，以使不同的激發圖型被 _ 引導至各通道之偵測區域。當細胞通過特定的通道之光束圖型 時，一系列有時間間隔的峰值將在所發射的螢光信號中產生。 因為細胞之流動速率或速度為已知，所以，各峰值之間的時 間、或頻率可用來確定哪個通道產生信號。制光學器件能夠 同時自所有的通道接收信號，且使用此技藝中已知的不同手段 (諸如’頻率調變/解調變），來分離及分析個別的通道信號。 099122308 17 201105969 此技藝中已知之各種方法可用以在 測區域内的激發光束。在-具體例中，:人=各通道之偵 他光偏移裝置之類的光學元件，可於相裝置或其 之基板中。該光學元件將均勻的光束轉換f則量晶片 内的空間_。在其他具體射，各通道之彳貞測區域 反射性材料，以找_型元件。刻或密有非 在其他具體例中，各種鏡 子、濾絲、或其他光阻或相位科段可时產生_。在盆 他具體例巾，諸如空間絲變料聲.光職器錢的主動元 件’可用來在各通道之彻m域内傳遞隨時間變化的空間強产 圖型。此等㈣圖型可以快於細胞通過偵測區域之行進的速^ 動態地改變。 圖4示意性例示一具體例，其中，激發光圖型係由平行於使 細胞404流過微流體裝置内諸平行流動通道4〇8、41〇及412 之偵測區域406的方向而間隔開的一系列條狀物4〇2所組成。 圖5例示分別針對通道408、410及412之樣品結果螢光信號 508、510及512。如所見，改變條狀物402之間的間隔，在針 對各通道而偵測的螢光信號中產生獨特的信號特徵，藉此，使 針對任何選定通道之偵測信號可與其他通道所產生的彳貞測信 號相分離。 圖6顯示另一具體例，其中，激發圖型元件602除了平行於 流向而改變外，亦垂直於流向而改變。如此使偵測光學器件可 採二維(與一維相反)方式接收關於細胞604的資料。使用細胞 099122308 18 201105969 604通過偵測區域時產生的二維螢光資訊，結合以圖塑元件 602之特定數學函數或形狀，可產生細胞之二維影像’藉此， &供更多關於細胞的資訊。 具有磁場分離器的微流體裝置： 本發明之某些具體例大體上而言係關於在微流體裝置上使 用細胞測量術(諸如’流動式細胞測量術或影像細胞測量術）而 使用電磁場來分離及分析生物樣品的系統。為提高細胞分選操 作之效率，需要以僅含有需要被研究或分離的彼等類型細胞的 樣品作為起始物。此技藝中已知之一種方法為在細胞測量分析 之刖將樣品離心。離心之後，樣品成分將被分離成多層。接著， 所需成分可容易地被萃取。然而，此舉引入在自初始收集容器 轉移至微流體裝置期間樣品層將變得再混合之可能性。藉由提 供與微流體裝置本身形成整體的分離能力，離心後分離層混合 之可能性降低。如此亦消除了對多個容器之需求，且降低了外 部污染物引入樣品中（或可能有害的樣品成分釋放於外部環境) 之可能性。 圖7示意性例示可用於達成含有生物樣品的所需生物成分 之分離的微流體裝置。在所例示之特定具體例中，來自細 胞供應源(未圖示)的細胞被輸入至輸入埠71〇，最初，在基板 702上之樣品孔72〇因電磁力而分離。此後，被分離的細胞在 分析區段川中經由細胞測量術而分析(在分析區段Ή2中發 生的特疋操作對於本發明而言不為關鍵的）。根據所進行的分 099122308 19 201105969 析之結果，細胞可分選至不同的腔室714令。出於簡單及易於 例示起見，圖7顯示單-通道在晶片观之諸組件、區域或區 段之間延伸。然而，應瞭解單一通道可為熟悉本技藝者可想到 的多個細胞測魏道及錄可能㈣道_之代表例。 在研究者或醫學專業人員所希望經由細胞測量術而分析的 細胞進入樣品孔720之前，除了將螢光分子附接至所需細胞之 外、或替代之，其可各自與磁性粒子減。根據系統所使 用的磁性粒子’可為次微米磁性珠粒或一般熟悉本技藝者可想 到的其他適當磁性粒子。來自輸入埠71〇的生物樣品經由流動 通道7〇8❿收容於樣品孔，中。相鄰的樣品孔72〇為可用於 在樣品孔中產生具有電磁力的電磁場的電磁體722。在一些具 體例中’電磁體722可為暫時性、半永久性、或永久性裝言狄 基板702板上(onboard)。在其他具體例中，使電磁體722鄰近 於樣品孔720,而未裝設於晶片观板上、附接至晶片7〇2、 或與晶片702連接。電磁體722可為獨立的，以使得至電磁體 的電能係經由使用電池或其他適當方式獲得。在其他具體例 中’電源線740被搞接至電磁體722,以向其提供電能。在某 二/、妝例中，用以提供磁場的電磁體722可永久性定位。當必 須建立磁場時，電磁體可以機械方式移入其位置，而且，當必 須移除磁場時移出其位置。 如上所述，耦接至磁性粒子的細胞724將進入電磁場，在電 磁力下被吸至電磁體722，且因此將被拉向鄰近底面7施的 099122308 20 201105969 樣品孔720之底部。在某些具體例中，樣品孔720與引導至分 析區段712的通道709，於鄰近底面72〇a的位置處呈流動連 通。在此等具體例中，f：初分離的細胞_ 724(經磁性粒子標記 的細胞)將流入通道709,且流入分析區段712中。為實現使細 胞724行進至通道709中，電磁體722之動力可暫時性、半永 久性、或永久性減小或消除，因而電磁力將相應地減小或消 除。因此，在其力量減小或消除之情況下’細胞724將自由行 進至引導至分析區段712的通道709中。在一些具體例中，磁 體722強到足以將細胞724固持於孔720之底部，直至磁力減 小為止。 在另一具體例中，磁體722不為電磁體，而事實上為永久磁 體。在此具體例中，磁體722產生的磁力可簡單地由樣品流體 流動（包括有進入通道709的細胞724)之力所克服，使細胞724 將離開樣品孔720。在其他具體例中，磁體722之物理位置或 磁強度被改變，以減小或消除足夠量之對細胞724之磁力，以 使細胞724將流出於孔720而流入引導至分析區段712的通道 7〇9中。在-些具體例中，讀722強到足以將細胞似固持 於孔720之底部，直至磁力減小為止。 另外’樣品孔720尚可選擇地包括有輪入埠，及廢料谭 732 ’以便分別引人及移出洗雜體。洗務流體可在進入細胞 測量分析之前，自樣品孔巾移料㈣有衫合需要的物質。 在此等具體财，當_越經料73()及732㈣過樣品孔 099122308 21 201105969 720時，洗滌流體可經由廢料琿732而將未吸至電磁體722的 生物樣品中之物質強推、推動、或引導出來。在某些具體例中， 來自生物樣品的未磁化物質將保持懸浮於樣品流體中，而且， 洗蘇流體移動通過樣品孔720之力，將使未磁化的物質離開該 孔。因此，未磁化的物質被阻止進入細胞測量分析，藉此，提 供在分析及分選之前的樣品純化之初始步驟。在此等具體例 中，將細胞724拉向孔720之底面72加的電磁力，大於洗滌 流體移動通過孔720的力，使細胞724不會經由廢料埠732被 洗出。 在被分離的細胞724在區段712中經分析後，諸細胞可通過 通道713，且基於諸細胞之不同特徵，選擇地被分選至不同的 孔或腔室714中。在某些具體例中，樣品孔714具有與其呈流 體連通的出口埠(未圖示）’以有助於自諸孔中移出被分選的樣 品。樣品流體可藉由適當控制導流器（flowdiverter)75〇而轉入 至孔714中。 在一具體例中，導流器750為壓電裝置，其可用電指令信號 致動，以便視導流器750之位置之不同而以機械方式將通過分 選通道713的液流導入孔714a或孔714b中。在其他具體例 中，導流器750不為壓電裝置，而可為例如自壁嵌入以使流向 偏轉的氣泡、藉由磁場移動或致動的偏流器、或如一般熟悉本 技藝者可想到的任何其他導流益或分選閘。 諸細胞可基於細胞之預期的未來用途而分選至不同的孔或 099122308 201105969 腔室714中。舉例而言，具有相同特徵或表型的細胞可分選至 個孔中，在έ亥孔中其被固定以供檢視，且分選至另一孔中， 而在該孔中其保持存活狀態以進行另外的功能測量。舉另一實 施例而S ’所需的細胞可分選至萃取孔或腔f中，*不合需要 的、、’田胞則可分選至廢料孔或腔室中。或者，諸細胞可基於體積 而’儿積至孔或腔室中，此與分選方法相反。晶片可包括此技藝 中已4的以物理方式將細胞自分析區段Μ〗導入腔室ΝΑ中的 手丰又或者’在分析完成之後，諸細胞可被促使離開晶片7犯。 與藉由啟_分析及預分選分離方法，系統700使研究者或醫 ^專業人貝可提高最終分選細胞之純度，此與健供分析及分 選的糸統相反。如此在經由細胞測量分析來分選所需的幹細胞 二有目為’生物樣品中的幹細胞之濃度一般為較低， 且其為極度所需的。 樣品孔720可採用任何適宜之實體形式，諸如，形成於基板 之表面中的可能為封閉、可能保持敞開、及/或可能包括蓋 子的孔。樣品孔720被顯示為接近晶片之頂部而定位；然而， 應瞭解’樣品孔可位於基板上的別處。 可堆疊的微流體裝置： 微流體 在某些具體例中，本發明大體上而言係關於可 壯 «X 「ι/·α. </ic ηχι. β、置呈可堆疊配置的微流體裝置，可有助於保護裳置本 ^於刮擦、磨損、或其他不期望有的損壞。根據本發明來堆System 200 can additionally include an excitation source 2〇6, a multi-core fiber optic 218, focusing lenses 212 and 220, and detection optics 226. The lenses 212 and 22 can be placed on the outer side of the substrate 202 as illustrated in Fig. 2, or can be mounted or integrally formed in the substrate 2〇2, depending on the needs of the particular application and cost considerations. Excitation source 206 can include a laser or other source of electromagnetic radiation known in the art. Detecting optics 226 can include various light sensing means known in the art, such as photomultiplier tubes, photodiodes, or collapsing photodiodes. In operation, an excitation source 206 (typically a laser) directs the beam toward a dichroic mirror 208. The dichroic mirrors are configured to reflect the difference in wavelengths received from the source 2〇6 099122308 15 201105969 to the wafer plus. Various other optical water focus assemblies, such as 'lens 212', can be used to ensure that the excitation source is properly focused on the read zone 210. When the sample cells 214 that traverse the microfluidic flow channel 204 pass through the detection zone 21 and emit fluorescence, the generated photons are emitted in multiple directions. - These photons will be emitted backwards towards the focusing lens 212, through dichroic mirrors 2〇8 (which are grouped to pass the wavelength of the fluorescent light), and enter the first end 2 of the multi-core fiber winding. Photons emitted from opposite sides of the wafer 2〇2 are selectively focused by the lens 220 and enter the second end of the fiber optic cable 218. The combined photon population is then emitted from the third end 224 of the optical ray 218 and received by the optical optics 226. Detection optics 226 can then collimate the combined emissions from the two fiber optic cores and transmit the transmission to - or a plurality of photodetectors (not shown) for measurement. In some embodiments, a one-way color and bandpass filter will be used to select a particular wavelength band and transmit it to the photodetector. In other embodiments, instead of a multi-core fiber optic cable, separate detection optics can be used to capture the emitted photons. Fig. 3 schematically illustrates another embodiment of the invention, shown here a system 3, wherein a plurality of light sources 306 are used to generate fluorescence in sample cells 314. In this embodiment, photons reflected and transmitted from sample cells 314 are detected from both sides of the cell measurement wafer 302. Again, the emitted photons will enter the ends 3丨6 and 322' of the fiber optic cable 318 and be captured by the detecting optics 326. In some embodiments, light source 306 can be grouped to produce light of different wavelengths. The detection optical device can then detect different types of cells of type 099122308 16 201105969 that have been stained and fluoresced at different wavelengths. Microfluidic devices using spatial intensity modulation: Some specific examples of the invention are generally directed to the use of intracellular measurements (such as flow cytometry or imaging cytometry) on microfluidic devices. The system of the sample. To increase total cell flux, multiple flow channels can be used in parallel to sort cells from a given sample. In such systems, the electronic processing device that detects the optics must distinguish between the signals received from the individual channels. This is accomplished by using individual excitation sources each having its own wavelength on each channel. However, this adds significant cost and complexity to the system. A single source can also be scanned across channels in a repeating manner, allowing the detection optics to determine which channel is scanned based on time. However, since it is possible for cells to pass through the detection area of the channel, and the light source excites the detection areas of different channels, this method also tends to be erroneous. As disclosed herein, another way to distinguish individual channel fluorescent signals is to direct single-excitation light to all channels 'however, change or shape the excitation beam pattern on (4) to make a difference The excitation pattern is directed by _ to the detection area of each channel. As the cell passes through the beam pattern of a particular channel, a series of time-interval peaks will be generated in the emitted fluorescent signal. Since the flow rate or velocity of the cells is known, the time, or frequency, between the peaks can be used to determine which channel produces the signal. The optics can simultaneously receive signals from all of the channels and separate and analyze individual channel signals using different means known in the art, such as 'frequency modulation/demodulation. 099122308 17 201105969 Various methods known in the art can be used to generate an excitation beam within the measurement region. In the specific example, the optical elements such as the person = the light redirecting means of each channel may be in the phase device or the substrate thereof. The optical element converts a uniform beam into a space _ within the wafer. In other specific shots, the reflective area of each channel is reflective material to find the _-type component. Engraved or dense. In other specific examples, various mirrors, filters, or other photoresist or phase segments can be generated. In his specific case, such as the space filament variable sound. The active element of the omnipotent money can be used to transmit time-varying spatially strong patterns in the m-domain of each channel. These (4) patterns can be dynamically changed faster than the speed at which the cells travel through the detection zone. Fig. 4 schematically illustrates a specific example in which the excitation pattern is spaced apart from the direction parallel to the detection region 406 that causes the cells 404 to flow through the parallel flow channels 4〇8, 41〇 and 412 in the microfluidic device. A series of strips consisting of 4〇2. FIG. 5 illustrates sample result fluorescent signals 508, 510, and 512 for channels 408, 410, and 412, respectively. As can be seen, varying the spacing between the strips 402 produces unique signal signatures in the fluorescent signals detected for each channel, thereby enabling detection signals for any selected channel to be generated with other channels. The signal is separated. Fig. 6 shows another specific example in which the excitation pattern element 602 is changed perpendicular to the flow direction in addition to being changed parallel to the flow direction. This allows the detection optics to receive information about the cells 604 in a two-dimensional (as opposed to one-dimensional) manner. The use of cells 099122308 18 201105969 604 to generate a two-dimensional image of the cell by combining the two-dimensional fluorescence information generated when detecting the region, combined with the specific mathematical function or shape of the patterned element 602, thereby providing more information about the cell. Information. Microfluidic Device with Magnetic Field Separator: Some specific examples of the present invention generally relate to the use of electromagnetic fields on a microfluidic device for separation using electromagnetic fields such as 'flow cytometry or imaging cytometry. And a system for analyzing biological samples. In order to increase the efficiency of cell sorting operations, it is necessary to use a sample containing only cells of the type that need to be studied or isolated as a starting material. One method known in the art is to centrifuge a sample after cell measurement analysis. After centrifugation, the sample components will be separated into multiple layers. Then, the desired ingredients can be easily extracted. However, this introduces the possibility that the sample layer will become remixed during transfer from the initial collection container to the microfluidic device. By providing a separation capability integral with the microfluidic device itself, the likelihood of separation of the separation layer after centrifugation is reduced. This also eliminates the need for multiple containers and reduces the likelihood of external contaminants being introduced into the sample (or potentially harmful sample components released to the external environment). Figure 7 is a schematic illustration of a microfluidic device that can be used to achieve separation of a desired biological component containing a biological sample. In the specific embodiment exemplified, cells from a cell supply source (not shown) are input to the input port 71, and initially, the sample holes 72 on the substrate 702 are separated by electromagnetic force. Thereafter, the isolated cells are analyzed by cytometry in the analysis section (the characteristic operation occurring in the analysis section Ή2 is not critical to the present invention). Based on the results of the analysis of the scores of 099122308 19 201105969, the cells can be sorted to different chambers 714. For simplicity and ease of illustration, Figure 7 shows a single-channel extension between components, regions or sections of the wafer. However, it should be understood that a single channel can be a representative example of a plurality of cell-measured Weis and the possible (four) channels that are familiar to those skilled in the art. Before the cells that the investigator or medical professional wish to analyze via cytometry enters the sample wells 720, they may be each reduced from the magnetic particles, in addition to, or instead of, attaching the fluorescent molecules to the desired cells. The magnetic particles used in accordance with the system' can be submicron magnetic beads or other suitable magnetic particles that are generally familiar to those skilled in the art. The biological sample from the input port 71〇 is accommodated in the sample well via the flow channel 7〇8❿. The adjacent sample well 72 is an electromagnet 722 that can be used to generate an electromagnetic field having electromagnetic force in the sample well. In some embodiments, the electromagnet 722 can be temporary, semi-permanent, or permanent on the substrate 702 onboard. In other embodiments, the electromagnet 722 is adjacent to the sample well 720 and is not mounted on the wafer viewing plate, attached to the wafer 7, or connected to the wafer 702. The electromagnets 722 can be self-contained such that electrical energy to the electromagnets is obtained via the use of a battery or other suitable means. In other embodiments, the power cord 740 is coupled to the electromagnet 722 to provide electrical energy thereto. In some second, makeup examples, the electromagnet 722 used to provide the magnetic field can be permanently positioned. When a magnetic field must be established, the electromagnet can be mechanically moved into its position and removed when it is necessary to remove the magnetic field. As described above, the cells 724 coupled to the magnetic particles will enter the electromagnetic field, be attracted to the electromagnet 722 under electromagnetic force, and will therefore be pulled toward the bottom of the sample hole 720 of the 099122308 20 201105969 adjacent to the bottom surface 7. In some embodiments, the sample well 720 and the channel 709 leading to the analysis section 712 are in flow communication adjacent the bottom surface 72A. In these specific examples, f: initially isolated cells _724 (cells labeled with magnetic particles) will flow into channel 709 and flow into analysis section 712. To effect travel of cell 724 into channel 709, the power of electromagnet 722 can be temporarily, semi-permanently, or permanently reduced or eliminated, and the electromagnetic force will be correspondingly reduced or eliminated. Thus, cell 724 will travel freely into channel 709 leading to analysis section 712 as its power is reduced or eliminated. In some embodiments, the magnet 722 is strong enough to hold the cell 724 at the bottom of the aperture 720 until the magnetic force is reduced. In another embodiment, the magnet 722 is not an electromagnet but is in fact a permanent magnet. In this particular example, the magnetic force generated by the magnet 722 can simply be overcome by the force of the sample fluid flow (including the cells 724 entering the channel 709) such that the cells 724 will exit the sample well 720. In other embodiments, the physical position or magnetic strength of the magnet 722 is altered to reduce or eliminate a sufficient amount of magnetic force to the cells 724 such that the cells 724 will flow out of the aperture 720 and flow into the channel leading to the analysis section 712. 7〇9. In some embodiments, the reading 722 is strong enough to hold the cells like the bottom of the well 720 until the magnetic force is reduced. Further, the sample well 720 optionally includes a wheel rim and a scrap 732 ' to introduce and remove the washing body, respectively. The wash fluid can be transferred from the sample well (4) to the desired material before entering the cell measurement analysis. In this particular case, when the _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Or lead it out. In some embodiments, the unmagnetized material from the biological sample will remain suspended in the sample fluid, and the force of the wash fluid moving through the sample well 720 will cause the unmagnetized material to exit the well. Therefore, the unmagnetized material is prevented from entering the cell measurement analysis, thereby providing an initial step of sample purification prior to analysis and sorting. In these particular examples, the electromagnetic force applied to pull the cells 724 toward the bottom surface 72 of the aperture 720 is greater than the force of the wash fluid moving through the aperture 720 so that the cells 724 are not washed out via the waste crucible 732. After the isolated cells 724 are analyzed in section 712, the cells can be selectively sorted into different wells or chambers 714 via channel 713 and based on the different characteristics of the cells. In some embodiments, sample well 714 has an exit port (not shown) in fluid communication therewith to facilitate removal of the sorted sample from the wells. The sample fluid can be transferred to the orifice 714 by appropriate control of the flow diverter 75 Torr. In one embodiment, the deflector 750 is a piezoelectric device that can be actuated by an electrical command signal to mechanically direct fluid flow through the sorting channel 713 into the aperture 714a or depending on the position of the deflector 750. In hole 714b. In other embodiments, the deflector 750 is not a piezoelectric device but may be, for example, a bubble that is embedded from the wall to deflect the flow, a deflector that is moved or actuated by the magnetic field, or as is generally known to those skilled in the art. Any other diversion or sorting gate. The cells can be sorted into different wells or 099122308 201105969 chambers 714 based on the intended future use of the cells. For example, cells with the same characteristics or phenotype can be sorted into wells, which are fixed for review in the wells and sorted into another well where they remain viable. For additional functional measurements. For another embodiment, the cells required for S' can be sorted into the extraction well or chamber f, and the 'field cells can be sorted into the waste well or chamber. Alternatively, the cells can be accumulated into the well or chamber based on volume, as opposed to the sorting method. The wafer may include a hand-practice that has physically introduced the cells from the analysis section into the chamber, or ' after the analysis is complete, the cells may be caused to leave the wafer 7. In conjunction with the separation and pre-separation separation method, the system 700 enables the researcher or the medical professional to increase the purity of the final sorted cells, as opposed to the health analysis and sorting system. Thus, the desired stem cells are sorted by cell measurement analysis. The concentration of stem cells in the biological sample is generally low, and it is extremely desirable. The sample well 720 can take any suitable physical form, such as a hole formed in the surface of the substrate that may be closed, may remain open, and/or may include a cover. The sample well 720 is shown positioned close to the top of the wafer; however, it should be understood that the sample well can be located elsewhere on the substrate. Stackable microfluidic devices: microfluids In some embodiments, the present invention is generally directed to a microfluid in a stackable configuration with respect to a strong X X / "α / · α. < / ic η χ .. A device that can help protect the skirt from scratching, abrasion, or other undesirable damage.

豐微流體震置，亦有助於益I 亦有助於猎由分隔裝置之前平面與後平面來防 099122308 23 201105969 止微流體裝魏在-起。此外，本文所揭示的具_有助於堆 豐具有-或多個不平表面的微流體裝置。舉例而言，—個表面 可具有附接至其上的透鏡’而該透鏡之—部分係延伸出基板表 面之平面。另外，可堆疊的配置可使研究者、醫學專業人員或 其他使用者較輕易地握持及操控裝置。其進一步可有助於使用 自動化及/或機态人元件來處理微流體裝置。 圖8不意性例示呈堆疊配置的微流體裝置8〇〇，諸如，細胞 測量晶片802。細胞測量晶片802 一般可被設計成使來自生物 樣品的物質可於其上經由細胞測量來分析（細胞測量分析中進 行的特定操作對於本發明而言不為關鍵的）。在某些具體例 中，細胞測S分析可包括例如影像細胞測量或流動式細胞測 量。根據所進行的分析之結果’樣品中之物質可選擇地分選於 裝設於晶片802上的一或多個不同的孔或腔室中。或者，在分 析完成之後，細胞可被促使離開晶片8〇2。應瞭解，晶片8〇2 之各種細胞測量組件及區段，出於簡單起見而未加以例示，且 可如一般熟悉本技藝者所想到作大幅改變。 在某些具體例中，細胞測量分析可對晶片8〇2之前表面或頂 面802a進行，且多個腿804可附接至晶片8〇2之背表面或底 面802b，以使晶片堆疊於彼此之上。在所例示之具體例中， 各晶片802 —般為矩形，且包括四條腿8〇4，自表面8〇2b向 下延伸並接近於該裝置之四個角而定位，如圖9中最佳例示。 然而，應瞭解，腿804在各晶片8〇2上之數目，可多於或少於 099122308 24 201105969 一般熟悉本技藝者可想到The abundance of microfluidic fluids also helps Yi. It also helps to prevent the front and rear planes from being separated by the separation device. 099122308 23 201105969 Moreover, the disclosed herein has a microfluidic device that facilitates stacking - or a plurality of uneven surfaces. For example, a surface may have a lens attached thereto and a portion of the lens extends out of the plane of the substrate surface. In addition, the stackable configuration allows researchers, medical professionals or other users to easily hold and manipulate the device. It may further facilitate the use of automated and/or physical human components to process microfluidic devices. Figure 8 is an illustration of a microfluidic device 8A, such as a cell measurement wafer 802, in a stacked configuration. The cell measurement wafer 802 can generally be designed such that the material from the biological sample can be analyzed via cell measurements (the particular operation performed in the cell measurement analysis is not critical to the invention). In some embodiments, the cytometric analysis can include, for example, image cell measurement or flow cytometry. Depending on the results of the analysis performed, the material in the sample is optionally sorted into one or more different wells or chambers mounted on wafer 802. Alternatively, after the analysis is complete, the cells can be caused to leave the wafer 8〇2. It will be appreciated that the various cell measuring assemblies and sections of wafer 8〇2 are not illustrated for simplicity and may vary widely as would be apparent to those skilled in the art. In some embodiments, the cell measurement analysis can be performed on the front surface 802a of the wafer 8〇2, and the plurality of legs 804 can be attached to the back surface or bottom surface 802b of the wafer 8〇2 to stack the wafers on each other. Above. In the illustrated embodiment, each of the wafers 802 is generally rectangular in shape and includes four legs 8〇4 extending downward from the surface 8〇2b and positioned adjacent to the four corners of the device, as best seen in FIG. Illustrative. However, it should be understood that the number of legs 804 on each wafer 8〇2 may be more or less than 099122308 24 201105969. It is generally understood by those skilled in the art.

四個’及/或可定位於晶片802上如— 的其他位置。在某些具體例中，腿8〇4 某些位置，以传| A g捡矗 堪ΟΛΛ η士 在所例示之具體例中，各腿_在_個末端義至表面祕Four 'and/or other locations on the wafer 802 such as - can be located. In some specific examples, the legs 8〇4 are in certain positions to pass | A g捡矗 ΟΛΛ η 在 在 In the specific example exemplified, each leg _ at the end of the _ end to the surface secret

外，所例示之腿8〇4實質上為具有圓形剖面形狀的圓柱形。各Further, the illustrated leg 8〇4 is substantially cylindrical having a circular cross-sectional shape. each

可具有不同於所例示者的形狀。舉例而言，腳部分8〇5可不存 在，而使得腿804在其整個長度上具有恆定的直徑。舉另一實 施例而言，804可包括正方形或矩形的剖面形狀。舉例而There may be shapes other than those exemplified. For example, the foot portion 8〇5 may be absent such that the leg 804 has a constant diameter throughout its length. In another embodiment, 804 can comprise a square or rectangular cross-sectional shape. For example

包括單一的矩形腳1004，如圖10中所例示。應瞭解，腿804、 1〇〇4僅為可併入微流體裝置中的一般熟悉本技藝者可想到的 眾多可能堆疊元件之兩個非限制性實施例。 具有3-D流體動力學聚焦以對齊精細胞的微流體裝置： 在某些具體例中，本發明大體上而言係關於一種微流體裝 置其月b夠進行3-D流體動力學聚焦，以使精細胞在該裝置中 之流動通道巾對齊。該裝置包括與精細賴行進的樣品細胞測 S通道相連接的鞘液(sheath fluid)次通道，該等次通道係定位 於兩個不同平面中，以使精細胞在樣品通道中以適當取向三維 099122308 25 201105969 地對齊，以供細胞測量分析。 圖η示意性例示形成於基板11〇2上的微流體裝置uo〇, 在其上方，自生物樣品(未圖示)輸入至輸入埠mo的諸如精細 胞之類的細胞’於區段1120中對齊(如下文將較詳細描綠），且 經由細胞測量術(諸如，流動式細胞測量術或影像細胞測量術) 而在分析區段1112中分析(分析區段1112中發生的特定操作 對於本發明而言不為關鍵的）。根據所進行的分析之結果，細 胞可選擇地分選至一或多個不同的孔或腔室1114中。在某些 具體例中，樣品孔1114具有與其流體連通的出口埠(未圖示）， 有助於自孔中移出經分選的樣品。樣品流體可藉由適當控制導 流器1116而導入至孔1114中。 在一具體例中，導流器1116為壓電裝置，其可用電指令信 號致動，以便視導流器1116之位置而定以機械方式將通過流 動通道的液流導入任一個孔1114中。在其他具體例中，導流 器1116不為壓電裝置’而可為例如自壁嵌人以使流向偏轉的 氣泡、藉由磁場移動或致動的偏流器、或如—般熟悉本技藝者 可想到的任何其他導流器或分選閘。 應瞭解’晶片11GG上所示的各種組件及區段，意欲以簡單 示意形式顯示細胞測量方法之操作，而且，晶片上之諸 細胞測量組件及區段可如-般熟悉本技藝者所想到作大 變。 田 細胞可基於細胞之不同特徵分選至不同腔室ΐιΐ4中。細胞 099122308 、 201105969 可基於細胞之預期的未來用途分選至不同腔室1114中。舉例 而言，具有相同特徵或表型之細胞可分選至一個腔室中，在該 腔室中被固定以供檢視’且分選至另一腔室中，而在該腔室中 保持存活狀態以供另外功能測量，或被適當地儲存以用作為部 • 分的基於細胞的治療程序。在一特定具體例中，被分選及分離 的精細胞可用於人類與動物之人工授精或體外受精。此等醫學 程序可在不孕症之情況下使用，且用以預防性別聯結的基因傳 播疾病（sex-linked gene-propagated disease)傳至下一代。舉另— 貫施例而言’所需的細胞可分選至萃取孔或腔室中，而不合需 要的細胞可分選至廢料孔或腔室中。或者，細胞可基於體積沉 積至腔室1114中，此與分選方法相反。或者，在分析完成之 後’細胞可被促使離開晶片1100。 出於簡單起見，圖11之例示圖顯示有兩個腔室1114;然而， 應瞭解，該微流體裝置可包括多於或少於兩個腔室，如一般熟 悉本技藝者可想到。另外，腔室1114被顯示為接近於基板11〇2 之底部而水平地對齊。然而，應瞭解，腔室1114若存在，則 可定位於基板1102上如一般熟悉本技藝者可想到的其他位 置。另外，出於簡單及易於例示起見，圖u顯示出單一通道 在基板11〇2之諸組件、區域或區段之間延伸。然而，應瞭解， 單-通道可為熟悉本技藝者可想到的多個細胞測量通道及多 種可能的通道組構之代表例。 圖12A及12B顯示可被引入至輸入痒111〇中以供細胞測量 099122308 27 201105969 分析之例示精細胞1104，苴由阁 _ . 、中圖12A顯示俯視圖，圖12B顯 不側視圖。在某些具體例中 _ 和、、，田肊包括頭1105及尾 1106。如所例不，一些精細 匕之頭通吊為薄餅形狀，其寬度W 大於八厚度T。出於細胞剛量 里刀衍炙目的，宜自頂部分析細 胞，如在圖12A中所定向，園立 口為更夕細胞經由其寬度W可用 於分析，所以，可得到更多資訊。 因此，可能需要對精細胞11n 曰、 也1104疋向，使得，在進入細胞測 I分析區段1112之前，全部官许w π 卩寬度W可用於分析。此定向作業 在細胞1104通過該裝置之分 刀析&奴1112時，提供了對細胞 之最均勻的照明。對於大多數哺乳動物物種，此定向作 業為獲得測量產生雌性的精細胞與產生雄性的精細胞之間的 舰差異所需的精準度如_。為實現此目標，晶片職 可包括流體動力學聚焦區段，諸如，對齊區段⑽，以適當 地定向精細胞1104(參看圖^ ι 、 锢口 13)。區段112〇可用於對齊自樣品 注入管1132進入樣品通道n 122的知細胞，而該注入管係與生 物樣品111Q相祕。如所例示，區段mo包括在同—平面、(諸 如’平面Υ)上的概進人次通道1124及1126、及在同一平面 (諸如，與平面Y正交的平面X)上的鞘液進人次通道1128及 113〇，以適當地對齊及定向流過樣品通道ιη2的精細胞。鞘 液進入次通道被組構成：自各個次通道進入樣品通道U22的 鞘液組合工作’以對齊及定向通道1122中之精細胞。在所例 示之特定具體例中，次通道1124及1126之剖面尺寸大於次通 099122308 28 201105969 道1128及1130,使得，可自彼等位置進入通道1122的鞘液之 量’大於自次通道1128及1130進入的鞘液之量。因此，在所 例不之特定具體例中，鞘液將使得精細胞以其較大寬度尺寸一 般落在X平面上而其較小厚度尺寸一般落在γ平面上的方式 對ί。另外’勒流次通道1124、1126、1128及1130亦可用來 將精細胞定位於通道112〇之中心，以更好地確保細胞通過可 為細胞測量分析之部分的光學系統之焦點。 適當定向係藉由使用具有斜面末端的管1132來增強，而該 斜面末端之平表面係與自次通道1124及1126進入的引入鞘流 對齊，或面向該引入鞘流。此系統將在自斜面管1132之末端 排出的精細胞之核芯樣品流與在管1132之末端上方通過的鞘 液之間產生層流。核芯樣品流在通過晶片11〇〇之分析區段 1112時，將保持不對稱，在X方向測量的寬度大於在γ方向 測量的高度。此不對稱性將使精細胞能夠在通過分析區段ιιΐ2 時保持被適當地定向。 應瞭解，區段1120可為基板1102之整體部分，或可以一般 熟悉本技藝者可想到的適當方式連接至基才反贈。在替代性 具體例中，區段112〇可與基板服分離，用—通道連接區段 1120之輸出部，連同基板11〇2上之流動通道。 具有音能耦合器的微流體裝置： 本發明之某些具體例大體上而言係關於一種微流體裝置，其 具有定位於該微流體裝置之組件中的音能輕合器，以破壞該組 099122308 _ 201105969 件中所含的細胞。細胞之破壞可使細胞内之内部分子物質釋 放’以使該物質可被研究者或醫學專#人M觀察到或測得。在 某些具體财’在細胞，分析(諸如，以流動式或影像細胞 測I術作為非限制性實施例)之後，細胞被分選至容器中。 圖μ示意性例示形成於基板1402上的微流體裝置14〇〇， 在/、上來自生物樣品(未圖示）的細胞或其他物質被輸入至輸 入埠1410，經由細胞測量術(諸如，流動式細胞測量術或影像 細胞測量術)在分析區段1412中分析(分析區段212中進行的 特定操作對於本發明而言不為關鍵的）。根據所進行的分析之 結果，細胞可選擇地分選至一或多個不同孔或腔室1414中。 在某些具體例中，樣品孔1414具有與其流體連通的出口埠(未 圖示），以有助於自孔中移出經分選的樣品。樣品流體可藉由 適當控制導流器1415而導入至孔1414中。 在一具體例中，導流器1415為壓電裝置，其可用電指令信 號致動，以便視導流器1415之位置之不同而以機械方式將通 過流動通道的液流導入任一個孔1414中。在其他具體例中， 導流器1415不為壓電裝置，而可為例如自壁嵌入以使流向偏 轉的氣泡、藉由磁場移動或致動的偏流器、或如—般熟悉本技 藝者可想到的任何其他導流器或分選閘。 應瞭解，圖14之例示圖中之基板1402上所示的各種組件及 區段’意欲以簡單示意形式顯示細胞測量方法之操作，而且， 基板1402上之諸細胞測量組件及區段可如一般熟悉本技蓺者 099122308 30 201105969 所想到作大幅改變。 細胞可基於細胞之不同特徵分選至不同孔或腔室1414中。 細胞可基於細胞之預期的未來用途而分選至不同的腔室1414 中。舉例而言，具有相同特徵或表型之細胞可分選至一個腔室 中’在該腔室中被固定以供檢視，且分選至另一腔室中，在該 腔至中保持存活狀態，以進行另外的功能測量，或被適當地儲 存以用作為部分的基於細胞的治療程序。舉另一實施例而言， 所需的細胞可分選至萃取孔或腔室中，而不合需要的細胞則可 分選至廢料孔或腔室中。或者，細胞可基於體積沉積至腔室 1414中，此與分選方法相反。晶片可包括如此技藝中已知的 以物理方式將細胞自分析區段1412導入腔室1414中的手段。 或者’在分析完成之後，細胞可被促使離開基板14〇2。 出於簡單起見，圖14之例示圖顯示有兩個腔室1414;然而， 應瞭解，該微流體裝置可包括多於或少於兩個腔室，如一般熟 悉本技藝者可想到。另外，腔室M14被顯示為接近於基板14〇2 之底部而水平地對齊。然而，應瞭解，腔室1414若存在，則 可定位於基板1402上如一般熟悉本技藝者可想到的其他位 置。另外，出於簡單及易於例示起見，圖14顯示單一通道在 基板1402諸組件、區域或區段之間延伸。然而，應瞭解，單 通道可為热悉本技藝者可想到的多個細胞測量通道及多種 可能的通道組構之代表例。 根據本發明，晶片1400上之至少一個組件包括音能耦合 099122308 31 201105969 器，以破壞該組件中所含的細胞。為例示實例，圖i5顯示了 呈定位於腔室1414 _探針之形式的音轉合器㈣。缺 而’應瞭解，音_合器可定位於晶片_之其他組件中，、，'、 包括在晶片上之輸人孔或其他細胞收集容[如所例示，音能 耦合器⑷6響應於自音能源！所接收的能量而振動^ 盡。應瞭解’耗合器⑷6可在腔室1414中以與所例示者不同 的方式訂定尺寸、成形、及/或另外組構及/或定向，如一般熟 悉本技藝者可想到。 … 在某些具體例中’具有懸浮細胞的樣品流體被收集於腔室 鼎内。音能輔合器㈣可用於響應於其自來源簡所接 收的音能，而在樣品流體内振動以破壞細胞。轉移至搞八器 ⑷6的音能可呈聲㈣式(諸如，料魏量），倾，音二處 理方法可應用於樣品流體’賜其巾獅細胞。在—些具體例 中，搞合器⑽可用於以其共振頻率(對應於其最大蝴振 動或振盡’以提供對細胞的最佳破壞。 在某些具體例中，細胞破壞具有足夠強度，以打碎細胞成 份’且將細胞之㈣分子物轉放於樣品流體巾。内部分子物 質接著可被研究者或醫料業人—麵_得。另外，在所 例示之具體例中，細胞在細胞測量分析及細胞分選至腔室⑷4 中之後被破壞。然而’應瞭解，輕合器可置於在細胞測量分析 之刖收谷細胞的不同晶片組件(諸如，初始的細 在細胞被破壞且分子物質釋放於初始的細胞貯存器Wl，分 099122308 32 201105969 子物質可被促使進入區段1412中，以供細胞測量分析。 在某些具體例中，音能源1420與耦合器1416分離，且可# 在晶片1400上對生物樣品進行細胞測量分析的外部儀器系統 合併。在其他具體例中，在完成細胞測量分析及自進行分析白勺 外部系統移除晶片1400之後，音能源1420可分別地施加於^ 片 1400。 具有產生相位變化的透鏡的微流體裝置： 本發明之某些具體例大體而言係關於在微流體裝置(諸如， 細胞測量晶片）上使用細胞測量術(諸如，流動式細胞測量術< 影像細胞測量術)來分析樣品的系統。為了在細胞測量操作期 間偵測或鑑別細胞，電磁輻射(諸如，可見光)之來源(諸如，雷 射器）被引導至偵測區。當細胞通過流動通道内之偵測區時， 光源使細胞發出螢光。此效應可如上文所述，藉由在細胞到達 4貞測區之前添加的某些染料來增強。 在某些應用中，可能有必要將各種光學元件併入裝置或晶片 之基板中’以改變各種表面之光散射性質。詳言之，諸如全息 照相元件或自由形式透鏡及鏡子之類的光成形光學器件，可用 來傳遞強度/相位/偏振空間圖型、或晶片各種表面上之其他光 偏移效應。因為此等元件可形成或蝕刻於晶片基板中，所以， 一旦初始模具或蝕刻程式已被設計，其複製成本極小。更特定 言之’用以聚焦入射激發光束的透鏡亦可製造成亦將在通道伯 測區域内傳遞特定的空間光圖型。 099122308 33 201105969 圖16示意性顯示一具體例，其中，聚焦透鏡1602被安裝於 微流體裝置16〇〇之基板1604之表面上。當樣品細胞1606通 過谓測流動通道16〇8時，激發光丨61〇聚焦於樣品細胞1606 上。聚焦透鏡1602之表面1612已被形成、蝕刻、或另外處理， 使其賦予入射的激發光161〇以空間上變化的相位改變。相位 改變轉換成為偵測區域上之光學全息照相效應，其可被產生以 形成特定的空間圖型或設計。 在製造聚焦透鏡1602期間，表面1612可塗有UV敏感材 料。脈波雷射接著可用來掃描表面1612，以曝光UV材料，且 根據所需圖型或影像而在表面產生小的變化。當聚焦透鏡16〇2 稍後被安裝於基板1604且接收激發光1610時，其將在偵測通 道1608内產生全息照相效應。在某些具體例中，聚焦透鏡16〇2 可完全由UV敏感材料製得，而非僅僅是表面ι612由UV敏 感材料製得。在其他具體例中，表面1612上之變化可藉由射 出成形或此技藝中已知的用於複製全息照相效應的其他手段 來達成。利用與全息照相元件相類似的方式，其他類型之自由 形式透鏡及鏡子形狀，亦可藉由首先使用此技藝中已知的微加 工技術(諸如，自電腦設計形狀進行鑽石車削)構築原圖(master copy)，而與聚焦透鏡1602組合。此原圖接著可使用射出成形 或此技藝中已知的其他手段來複製。 特定的圖型或所需的全息照相效應可針對各種用途而選 擇。舉一個非限制性實施例而言，全息照相透鏡或自由形式光 099122308 34 201105969 學元件，可用來產生空間上變 化的圖型，以確定偵測通道之獨 特的特徵」㈣爾e)。·—缝量，㈣流動通道 經常在細胞測量裝置(諸如， ’同式、，、田胞分選儀或微流體裝置）中 平行使用，以測量及/或分】g & 飞刀建細月包。在此等系統中，僧測光學 "件及相關電子處理設備必須能區分自個別的通道所接收的 信號。實現此舉之—種方式為將單—激發光源同時引導至所有 通道’但，在空間上改變或成形激發光束圖型，以使不同的激 發圖型被”至各通道之偵_域。當細胞通過特定通道之光 束圖型時’-㈣树„_峰值將在所飾的螢光信號中 產生。上文討論之相位變化透鏡在此情況下可用來產生全&昭 相效應。全息_效應將均勻的光束轉_各通道之偵測區域 内之獨特的空間圖型。 具有透射鏡及反射鏡的微流體裝置： 在先前技術系統巾’統被施加於細關量裝置之一側，價 測光學器件亦僅偵測來自樣品細胞之—側的螢光。透射鏡或其 他光學器件經常用來將發射光子之—部分聚焦於彳貞測光學器 件，然而，如此仍限制自螢光樣品細胞所捕獲的光子之量，因 為，在相反方向發射的光子並未被透射鏡捕獲。激發光強度可 提高，以產生更多以螢光方式產生的光子，然而，如此亦增加 信號中雜訊之量。另一解決方案為使用設置於不同位置的多組 偵測光學器件，但，如此增加了大量費用。 為了在單一偵測光學器件情況下儘可能多地捕獲由發螢光 099122308 35 201105969 樣品細胞產生的光子，反射鏡可置於樣品細胞之與偵測光學器 件相反的一側。反射鏡將捕獲大量未被透射鏡所捕獲的光子， 將其往回反射通過透射鏡，且進入偵測光學器件。如此使得該 系統對於既定激發光強度捕獲更多發射光子，而不增加接收信 X 號中之雜訊，且不花費多個偵測光學器件之成本。 圖π示意性例示使用細胞測量術來分析樣品的系統17〇〇。 系統Π00可包含形成於基板1702上的微流體細胞測量裝置 (在此處以側視圖顯示），其具有含納於其中的偵測流動通道 1704。出於簡單及易於例示起見，圖17顯示基板17〇2内之單 一通道。然而，應瞭解，單一通道可為熟悉本技藝者可想到的 多個細胞測量通道及多種可能的通道組構之代表例。各種其他 細胞測量組件亦可含納於基板17〇2上’但對於本發明而言不 為關鍵的。 系統1700可另外包含透射鏡π〇6、反射鏡1707、激發光源 1710、二向色(dichroic)鏡1712、及偵測光學器件1714。透射 鏡1706及反射鏡1707視特定應用及成本考慮之不同而可安裝 或整體式形成於圖17所例示的基板1702内，或可置於基板 1702之外侧。激發光源πΐ〇可包含雷射或此技藝中已知的其 他光源。偵測光學器件1714可包含此技藝中已知的各種感光 手段’諸如，光電倍增管。 在操作中，激發光源1710將光束引向二向色鏡1712。二向 色鏡被組構成將由激發光源1710所接收的波長反射至偵測通 099122308 36 201105969 道1704内之偵測區1716。當樣品細胞1718通過偵測區1716 且發出螢光時，所產生的光子在多個方向上發射。一些光子將 朝聚焦透鏡Π06發射回來，通過二向色鏡1712(其被組構成使 螢光波長通過)且被偵測光學器件1714所感應。自晶片1702 之相反側發射的光子將被反射鏡1707捕獲，引導回至透射鏡 1706，且聚焦於偵測光學器件1714。 透射鏡1706、偵測通道1704、及反射鏡1707之間的相對距 離被設置成：自反射鏡1707反射的光亦被透射鏡1706適當地 聚焦於偵測光學器件1714。舉例而言，反射鏡1707可能需要 較靠近偵測通道1704。當透射鏡1706及反射鏡1707被直接 安裝於基板1702上時，如此可藉由偏置如圖17中所示之偵測 通道1704 ’以確保透射鏡及反射鏡相對於偵測通道π〇4適當 置放而實現。 具有整體式形成的光學器件的微流體裝置： 各種光學裝置(諸如，透鏡)經常設置於微流體裝置中之入射 及/或射出光束之路徑中，以提供最大量之所觀察到的樣品細 胞之光子傳遞及回收。此等光學裝置亦經常需要改變由偵測區 域所發射的光之數值孔徑(NA)，以使其可適當地被光纖纜線接 收。此為必要的，因為，發射光之ΝΑ通常很高，而光纖纜線 之ΝΑ通常很低。若發射光之ΝΑ在到達光纖纜線之前未減 小’則僅有一小部分之發射光被成功傳送通過光纖纜線。 然而，一個問題在於：各種入射及射出光束在通過光束路句 099122308 37 201105969 :之各種材料時會發生折射。舉例而言，當聚焦的激發束自空 氣穿入微流體“之純(其具有正常平面）時，光束將以某種 程度折射，使其較難以將光束聚焦於晶片内之預期的㈣區 域。為克服此問題，聚焦透鏡或其他光學科可形成於晶片基 本身田中因為*焦光學器件與晶片基板之間的氣隙被消 除’且因為m學科及晶#基板係自同—件材料之一部分 所形成且保留同-件材料之-部分，所以，消除了晶片材：内 光束之不期望有的折射。 圖18例示使用細胞測量術來分析樣品的系統18〇〇。系統 1800可包含微流體裝置基板(其側視隨段顯示於該圖 中）’其具有含納於其中的償測通道腦。出於簡單及易於例 不起見，ϋ 18顯示細胞測量晶片臓内之單一通道。然而， 應瞭解，單-通道可為熟悉本技藝者可想到的多個細胞測量通 道及夕種可旎的通道組構之代表例。各種其他細胞測量組件亦 可含納於如上文所述之細胞測量晶片18〇2上，但對於本發明 而言不為關鍵的。 基板1802另外包含此處顯示為聚焦透鏡的光學器件18〇6。 光學器件1806整體式形成於基板18〇2之基板中。此節可藉由 此技藝中已知的各種方法而實現。舉例而言，光學器件18〇6 可形成於用來製造基板1802的射出模具中。在其他具體例 中，在基本晶片形狀形成之後，光學器件1806可被加工至晶 片基板中’或另外在晶片基板外成形。 099122308 38 201105969 在知作中，激發光源1810將光束引向二向色鏡1812。二向 色鏡1812被組構成：將自激發光源181〇接收的波長反射至偵 測通道1804内之偵測區域1814。此技藝中已知的各種其他光 學部件可用來將激發光束彳丨導至光學^件18Q6，其細節對於 本發明而吕不為關鍵的。當樣品細胞1816通過偵測區域1814 且被激發光束促使發出螢光時，所產生的光子將被光學器件 1806收集或聚焦，通過二向色鏡1812(其被組構成使螢光波長 通過），且最終被偵測光學器件1816所偵測。偵測光學器件 1816可包含此技藝中已知的各種感光手段，諸如，光電倍增 官。在某些具體例中，此技藝中已知的其他組件(諸如，光纖 纜線)可用來將發射光束導引至偵測光學器件1816。 應瞭解，光學器件1806可包含供聚焦、傳播、濾光、或以 其他方式處理被價測區域1814内之樣品細胞1816所接收、戈 由其產生的光束的任何光學元件。另外，多個光學元件可安枣 於基板1802上。舉一個非限制性實施例而言，具有反射膜的 另外的光學器件1818可形成於基板1802之相反表面182〇 中’以將另外的光子引導回至偵測光學器件1816。在某些鹿 用中，宜於在表面1820中形成呈侵入(與突起相對)形式的光 學器件1818，如圖18中所例示。如此使得基板18〇2之表面 1820可與安裝裝置(未圖示）内之相應平面齊平置放。 具有非整體區塊選擇性試劑傳遞結構的微流體裝置： 在許多情況下，可能需要添加試劑，以在執行細胞測量分析 099122308 39 201105969 之前活化或另外製備樣品。所需的試劑可在製造 流體裝置中，從而消除了使用者在使用時添加之=至微 如此則要求許多不同類型之裝置要被製造商維持=然而’ 置僅適用:少數應用。為提高製造效率及降低庫存：本而： ，而_的結構可在製造之後與微流 造商可製造出可接受許多不同試劑傳 遞結構的早一微流體裝置。 圖19例㈣組構魏容分綱觸結構赚的微流體裝置 觸。裝置測可包含藉峨流體流動通道19()8而連接至試 劑收容部分1906的細胞測量分析部分19G4。裝置簡亦包 含樣品收料1910,以·容待分_樣心樣純容淳测 藉由微流體流動通道19!2而連接至細胞測量分析部分19〇4。 在某些具體例中’在樣品添加至裝置19GG之前，試劑傳遞結 構1902與試劑收容部分腿相配i此配合可由使用者在使 用時或由製造商在載運裝置之前進行 '細胞測量分析部 分1904對所收容的樣品及試劑混合物進行細胞測量分析。舉 例而言’細胞測量分析區段可將所需的細胞分選至萃取 孔1914中，且將不合需要的細胞分選至廢料孔1916中。細胞 測量分析區段1904中進行的特定操作、及微流體通道之特定 導引，對於本發明而言不為關鍵的。 試劑收容部分1906被製造成具有配置成栅格或其他標準化 佈局的複數個收容孔1918。各種試劑傳遞結構丨9〇2接著可被 099122308 40 201105969 製造出來，其具有試劑部分1920 ’當試劑傳遞結構1902與試 劑收容部分1906相配合時，該等試劑部分1920在對應於至少 一個收容孔1918的位置含有試劑。在某些具體例中，試劑傳 遞結構1902中特定試劑類型之位置將亦被標準化。舉例而 • 言’在對應於特定收容孔1918的位置處的任何試劑，總是同 一類型的。因為如此，用以將試劑施加於試劑傳遞結構19〇2 的設備’在製造期間可被最優化成：若試劑正被施加於特定試 劑傳遞結構19〇2，則總是將某一試劑類型施加於同一位置。 如此亦使得細胞測量分析區段1904及各種微流體流動通道 1908可基於所需試劑之已知位置來設計及最優化。 試劑傳遞結構麗可由此技藝中已知的適用於儲存試劑的 任何材料所製成。舉例而言，試劑結構1902可呈係帶或其他 可繞性條帶之形式…旦施加於觸收容部分1906，即以所 施加的黏著劑將試劑結構19〇2固持於適當位置。在某些具體 例中，試劑收容部分懸可呈凹陷於裝置·中，其試劑傳 遞、構1902係由剛性材料製成’且被訂定尺寸成為隱蔽地配 •接於試劑收容部分懸内，從而確保適當地置放試劑傳遞結 構聰。收容孔1918亦可包含可刺穿試劑部分咖而將試 劑部分1920内所含的試劑釋放或暴露於收容孔⑼8及通道 1908中的針。 Θ ’員示另具體例’其中，試劑結構2GG2上所含納的試 劑聊係藉由可溶的障壁2_而固持於結構聰内。活化 099122308 41 201105969 材料2045被包括於裝置2000内之試劑收容部分2006上。一 旦試劑結構2002被施加於試劑收容部分2〇〇6，活化材料2〇45 即致使可溶障壁2040溶解，從而將試劑2020釋放於試劑收容 孔2018及流動通道2〇〇8中。在某些具體例中，由外部來源所 施加的聚焦雷射束或音能，可用以致使可溶障壁2〇4〇溶解。 在其他具體例中’可溶障壁2〇4〇及/或活化材料2〇45可由溫 度敏感材料製成，藉此，在將試劑結構2〇〇2配合至裝置2〇〇〇 之後升高溫度’將致使可溶障壁2〇4〇溶解。顯示此溫度敏感 特性的各種材料在此技藝中為已知。舉例而言’試劑可設置於 瓊脂糠中’其將保持細胞含於試劑内’直至瓊脂糖被加熱溶解 為止。在某些具體例中，可溶障壁2〇4()可併有微電機結構 (MEIVISMS ° 在流動通道中併有光波導器的微流體裝置： 本發明之某些具體例大體上而言係關於微流體裝置，諸如， 細胞測量晶片，其具有被組構成光波導器以更好地引導照明光 線通過通道的細胞測量通道。在某些具體例中，使細胞測量通 道適合成為光波導器，包括：選擇具有所需折射率的材料來形 成通道壁。另外，光波導通道之效用亦可視通過通道的流體之 折射率而定。儘官本發明之波導通道在本文中出於簡潔起見被 論述為引導可見光，但，光僅為可由波導通道引導的眾多可能 類型之電磁輻射之一個非限制性實施例。 圖21示思性例示出系統2100，其在微流體裝置上具有例示 099122308 42 201105969 性細胞測量通道2102，作為該裝置所進行的細胞測量分析(諸 如，流動式細胞測量或影像細胞測量)之部分(特定細胞測量分 析操作對於本發明而言不為關鍵的）。如所例示，通道21〇2具 有構成兩個實質上垂直的區段21〇2a及2102b的角2103。系 • 統2100沿著通道區段2102a包括有發光系統2106及光收集系 統2107，而光係用以照明細胞，以供作為細胞測量分析之— 部分的偵測。在所例示之實施例中，光收集系統21〇7被定位 成其光軸與通過通道區段2102a的液流之軸線對齊，且發光系 統2106被定位成其光軸與通過通道區段21〇2的液流軸線呈正 交。應瞭解，系統2106及2107之特定組構、位置及操作，必 須能將輻射正確地耦合於系統中，但，對於本發明而言不為關 鍵的，該等系統為一般熟悉本技藝者可想到的適當系統。生物 樣品通過通道2102之整體液流，係由通道内標示的大頭箭頭 2120所例示。由系統2106發射的光在整個通道21〇2中被内 部地反射至收集系統2107，且整體以箭頭2122表示。因此， 通道2102可充當光波導器。 _ . 通道2102接收來自發光系統2106的照明光，且將其沿著通 道區段2102a之長度傳輸，而朝向光收集系統21〇7發射出去。 為實現此舉，通道2102之通道壁2104至少沿著區段2l〇2a可 具有適當的折射率，使其在光通過區段2102a時，内部地反射 光。另外，通道壁2104沿著區段2102a之長度之全部或部分 可被遮蔽(例如，用反射塗層），使光僅在所需的位置發射。此 099122308 43 201105969 種配置可允許通道僅在所需的區域發光，其可為連續或多點發 射。 在某些具體例中，通道2102可包括與發光系統21〇6對齊的 壁區段2108、及與光收集系統2107對齊的壁區段2109，以允 許光發射至通道中及發射出通道。另外，在某些具體例中，壁 區段2108及2109可由具有與壁2104之其餘部分不同的折射 率的不同材料所構成，該不同材料及折射率被設計成允許光經 由其間而發射。舉另一實施例而言’在通道壁2104被遮蔽的 具體例中，壁區段2108及2109可保持不被遮蔽，以使光可通 過。 在流動通道中併有光波導器及反射面的微流體裝置： 在某些具體例中’反射面可定位於光波導通道之一個末端， 以進一步有助於引導照明。圖22示意性例示出系統2200，其 在微流體裝置上具有含通道壁2204的例示性細胞測量通道 2202，作為對該裝置進行的細胞測量分析(諸如，流動式細胞 測里或影像細胞測5 )之一部分（特定細胞測量分析操作對於 本發明而言不為關鍵的）。如所例示，通道2202在實質上垂直 的通道區段2202a與2202b之間具有角2210，且在實質上垂 直的區段2202b與2202c之間具有角2203。另外，通道區段 2202b整體地包括鄰近於角2210的第一末端2240、及鄰近於 角2203的第二末端2242。系統2200沿著通道區段22〇2b包 括發光系統2206及光收集系統2207，而光係用以照明細胞， 099122308 44 201105969 胞測量分析之—部分的偵測。在所例示之實施例 的液济ιΓ錢22G7蚊位成其妹與通料道®段22〇2b 通道㈣齊’且發輕統22G6被跋成其光軸與通過 的液流之鱗呈正交。應瞭解，系統及 轉中讀構、㈣及操作，必須⑽騎正確地輕合於 對於本發明而言不為_的，該等系統為一般熟 心本技藝者可想到的適當系統。 另外具有鏡面迎的諸如鏡子223〇之類的反射面，被定 位成鄰近於整體上與光收集系統纖相對置的第一末端 2240。如所㈣，鏡子2⑽可與猶壁纖構絲體。鏡子 可為二不的球形，或其可具有適於將通道波導器所耦合的發射 光沿:通道2202向下反射回來的其他形狀。在其他具體例 中銃子2230可為可變形的鏡子，其在該領域中一般稱作自 ^Wi#(adaptive Qptie)。此錄子可使其雜在反饋系 統控制下快速調節。 外生物樣品通過通道22〇2之整體液流，由通道内標示的大頭 ^頭2220例示。自系統2206發射的光在整個通道2皿中被 内。p地反射至收集系統2207,且整體以箭頭2222表示。因此， 通道2202可充當光波導器。 通道2202接收來自發光系統22〇6的照明光，且將其沿著通 道區段2202b之長度傳輸，而朝向光收集系統22〇7發射出去。 為實現此舉’通道2202之通道壁2204至少沿著區段22〇2b可 099122308 45 201105969 具有適當的折射率，使其在光通過區段2202b時，内部地反射 光。另外，通道壁2204沿著區段2202b之長度之全部或部分 可被遮蔽(例如，用反射塗層），使光僅在所需的位置發射。此 種配置可允許通道僅在所需的區域發光，其可為連續或多點發 射。此外，球面的鏡面2232被定位成將可在末端224〇處到達 鏡子2230的光之任何部分，在末端2242處朝向光收集系統 2207反射回來。使用鏡子2230，藉由防止自系統2206發射的 光傳輸來到與系統2207相對立的末端2240，來改良光波導通 道之效率及效用。 在某些具體例中’通道2202可包括與發光系統2206對齊的 壁區段2208、及與光收集系統2207對齊的壁區段2209，以允 許光發射至通道中及發射出通道。另外，在某些具體例中，壁 區段2208及2209可由具有與壁2204之其餘部分不同的折射 率的不同材料所構成，該不同材料及折射率被設計成允許光經 由其間而發射。舉另一實施例而言’在通道壁2204被遮蔽的 具體例中，壁區段2208及2209可保持不被遮蔽，以使光可通 過。 用於病毒偵測及分選的微流體裝置： 本發明之某些具體例大體上而言係關於偵測身體樣品中之 病毒粒子，其係藉由在諸如細胞測量晶片之類的微流體裝置上 對樣品進行細胞測量分析、及將病毒粒子分選至裝設於該晶片 上的孔或腔室中來達成。細胞測量分析可為流動式細胞測量分 099122308 46 201105969 析或影像細胞測量分析。分選來自身體樣品的病毒粒子，使研 究者或醫學專業人員可捕獲病毒群，以供進一步觀察或測試。 如本文中所用，術語「細胞測量」（Cyt〇metry)被廣泛使用，且 意欲包括測量任何適當物質，包括細胞、及/或病毒粒子，作 為兩個非限制性實施例。 系統2300(如23圖中示意性例示）包括形成於基板23〇2的微 流體裝置，其允許在細胞測量分析期間偵測病毒粒子，.及在分 析後分選病毒粒子。作為系、統2300之一部分，身體樣品(未圖 示)被輸入至輸入埠2310，在分析區段2312中之基板23〇2上 經由細胞測量術分析(分析區段2312中進行的特定操作對於本 發明而言;^為_的Η罐所進行的分析之結果，輸入至輸 入埠23U)而在分析區段2312中被偵測的身體樣品中之病毒: 子，可被分選至-或多健室加4中。在某些具體例中，樣 品孔23Μ具有與其流體連通的出口璋(未圖示），以有助於自 孔中移出經分選的樣品。樣品流體可藉由適當控制導流器咖 而導入至孔2314中。 你一具肢例中，導流器遍為壓絲置，其可用電指抑 =動’以便視導流器腿之位置之不同而以機械方式㈣ =動通道的液流導入任-個孔2314中。在其他具體例中 為壓電裝置’而可為例如自壁嵌入以使_ =:猎由磁場移動或致動的偏流器、或如_般熟綱 我者可心到的任何其他導流器或分選閘。 099122308 201105969 病毒粒子可基於粒子之預期的未來用途進一步分選至不同 的孔或腔室2314中。舉例而言，具有相同特徵的病毒粒子可 分選至腔室2314a中，在該腔室中被固定以供藉由電子顯微鏡 觀察’且分選至腔室2314b巾，而在該腔室中與被研究藥理學 功效的化學物質反應’其餘樣品則根據其他準則被分選至腔室 2314c及2314d中。在其他具體例中，所有的腔室均可 收容分析區段2312中偵測的病毒粒子。晶片23〇〇可包括如此 技藝中已知的以物理方式將病毒粒子自分析區段2312導入腔 室2314中的手段。或者，身體樣品中之物質可用來在分析完 成之後離開晶片2300。 在所例示之具體例中，存在四個例示的腔室2314 ;然而， 應瞭解，可存在多於或少於四個腔室，如熟悉本技藝者一般可 想到。具有複數個腔室2314的基板2302可被設計成使預定量 之病毒粒子被分選至各腔室中。出於便於例示起見，腔室2314 被例示為水平地對齊，但，亦應瞭解，該等腔室可以其他方式 定位於晶片上’如熟悉本技藝者一般可想到。另外，出於簡單 及易於例示起見，圖23顯示單一通道在基板2302之諸組件、 區域、或區段之間延伸。然而，應瞭解，單一通道可為熟悉本 技藝者可想到的多個細胞測量通道及多種可能的通道組構之 代表例。 在輸入埠2310輸入的身體樣品可為血液樣品、尿液樣品、 組織樣品、唾液樣品、細胞樣品、或其組合，此僅列出一些非 099122308 48 201105969 限制性實施例。除偵測及分選身體樣品中任何病毒粒子之存在 外’系統2300亦可被組構成測量樣品中之病毒粒子數。在某 些具體例中，在區段2312之細胞測量分析能夠測量樣品中之 病毒粒子數。在其他具體例中，病毒粒子被分選至基板23〇2 上之腔室2314中，且可在分選之後被測量或計數。 本發明之某些具體例大體上而言係關於藉由在諸如細胞測 量晶片之類的微流體裝置上對樣品進行細胞測量分析，來偵測 (而非刀遥)身體樣品中之病毒粒子。細胞測量分析可為流動式 細胞測量分析或影像細胞測量分析。由於在微流體裝置上測試 之文控性質的緣故，使用該裝置以經由細胞測量術來偵測病毒 粒子，提供了增加的安全性及降低的暴露於研究者或醫學專業 人員的風險。另外，微流體裝置使細胞測量分析可以相對缓慢 的速率進行，從而使小病毒粒子可得到偵測。此外，微流體裝 置亦允許經由一個測量裝置同時偵測細胞與病毒粒子。如本文 中所用，術語「細胞測量」被廣泛使用，且意欲包括測量任何 適當物質，包括細胞、及/或病毒粒子，作為兩個非限制性實 施例。 在某些具體例中，病毒粒子可利用識別RNA中分子之特定 序列的特定螢光探針予以標記。細胞測量儀器系統可基於光散 射來偵測病毒粒子之存在，且基於螢光發射之強度及波長而將 該粒子分類。在一些具體例中，在同一生物樣品中，多種螢光 探針可被使用，而且，多種病毒可被偵測。 099122308 49 201105969 除_在輸入埠2310輸入的身體樣品中有任何病毒粒子之 存在外’系統230()亦可被組構成測量樣品中之病毒粒子數。 在某些^體例中，在區段2312之細胞測量分析能夠測量樣品 中之病毒粒子數。在其他具體财，病毒粒子可被分選至基板 2302上之孔或腔室中，且可在分選之後_量或計數。在涉 及AIDS賴的具體例巾’身體樣品巾每單位血液之病毒粒子 數可經由細胞測量分析來測量。計算絲粒子數(亦稱作病毒 負荷)可為評估AIDS患者之健康中的重要測量。若在區段2312 進行分析之後樣品保留於基板 i，則處理樣品之安全性 大幅改良’因為’整個晶片測可如任何其他被污染的醫學 裝置(例如，如用過的注射器）般被拋棄。 顯不顏色改變以指示使用或結果的微流體裝置： 本發明之某些具體例大體上而言係關於一種微流體裝置，諸 如，細胞測！_晶片，其能夠在細胞測量分析之後提供可見的指 不，以指示細胞測量分析之結果。細胞測量分析可為如上所述 之流動式細胞測量分析、或影像細胞測量分析。在一例示性具 體例中，若分析產生陽性結果，則可見的指示為在分析完成後 由染料淹沒裝置之組件所引起的顏色改變。 圖24例示出系統2400，其中，來自於細胞供應源(未圖示） 的細胞被輸入至基板2402上之輸入埠2410中，且經由細胞測 量術在分析區段2412中分析(分析區段2412中發生的特定操 作對於本發明而言不為關鍵的）。根據所進行的分析之結果，A single rectangular foot 1004 is included, as illustrated in FIG. It will be appreciated that the legs 804, 1〇〇4 are only two non-limiting embodiments of numerous possible stacked components that are generally incorporated into the microfluidic device and are generally contemplated by those skilled in the art. Microfluidic device with 3-D hydrodynamic focusing to align sperm cells: In some embodiments, the present invention generally relates to a microfluidic device that is capable of 3-D hydrodynamic focusing for a month The sperm cells are aligned in the flow channel in the device. The apparatus includes a sheath fluid secondary channel coupled to the sample cell S channel of the finely-traveled sample, the secondary channel being positioned in two different planes to allow the sperm cells to be properly oriented in the sample channel. 099122308 25 201105969 Ground alignment for cell measurement analysis. Figure η schematically illustrates a microfluidic device uo 形成 formed on a substrate 11 〇 2 above which a cell (such as a sperm cell) that is input from a biological sample (not shown) to the input 埠mo is in the segment 1120 Alignment (green will be described in more detail below) and analyzed in analysis section 1112 via cytometry (such as flow cytometry or imaging cytometry) (specific operations occurring in analysis section 1112 for this It is not critical to the invention). Based on the results of the analysis performed, the cells are optionally sorted into one or more different wells or chambers 1114. In some embodiments, the sample well 1114 has an outlet port (not shown) in fluid communication therewith to facilitate removal of the sorted sample from the well. The sample fluid can be introduced into the well 1114 by appropriate control of the deflector 1116. In one embodiment, the deflector 1116 is a piezoelectric device that can be actuated by an electrical command signal to mechanically direct fluid flow through the flow channel into either of the apertures 1114 depending on the position of the deflector 1116. In other embodiments, the deflector 1116 is not a piezoelectric device' but may be, for example, a bubble that is embedded from the wall to deflect the flow, a deflector that is moved or actuated by the magnetic field, or as familiar to those skilled in the art. Any other deflector or sorting gate that is conceivable. It will be appreciated that the various components and sections shown on wafer 11GG are intended to show the operation of the cell measurement method in a simple schematic form, and that the cell measurement components and sections on the wafer can be as familiar to those skilled in the art. Great change. Field cells can be sorted into different chambers based on different characteristics of the cells. Cells 099122308, 201105969 can be sorted into different chambers 1114 based on the intended future use of the cells. For example, cells having the same characteristics or phenotype can be sorted into one chamber where they are fixed for viewing 'and sorted into another chamber while remaining alive in the chamber The status is measured for additional function or is stored appropriately for use as a partial cell-based treatment procedure. In a specific embodiment, the sorted and isolated sperm cells can be used for artificial insemination or in vitro fertilization of humans and animals. These medical procedures can be used in the case of infertility and spread to the next generation to prevent sex-linked gene-propagated disease. Alternatively, the desired cells can be sorted into extraction wells or chambers, and undesirable cells can be sorted into waste wells or chambers. Alternatively, the cells may be deposited into the chamber 1114 based on volume, as opposed to the sorting method. Alternatively, the cells can be caused to leave the wafer 1100 after the analysis is complete. For simplicity, the illustration of Figure 11 shows two chambers 1114; however, it should be understood that the microfluidic device can include more or less than two chambers, as will be appreciated by those skilled in the art. Additionally, the chamber 1114 is shown as being horizontally aligned proximate to the bottom of the substrate 11〇2. However, it should be understood that the chamber 1114, if present, can be positioned on the substrate 1102 as would be otherwise familiar to those skilled in the art. Additionally, for simplicity and ease of illustration, Figure u shows a single channel extending between components, regions or sections of substrate 11〇2. However, it should be understood that the single-channel can be a representative of a plurality of cell measurement channels and a variety of possible channel configurations that are familiar to those skilled in the art. Figures 12A and 12B show an exemplary sperm cell 1104 that can be introduced into the input itch 111 供 for cell measurement 099122308 27 201105969 analysis, a top view of Figure 12A and a side view of Figure 12B. In some specific examples, _ and , ,, 肊 include head 1105 and tail 1106. As an example, some of the fine heads are hoisted into a pancake shape with a width W greater than eight thicknesses T. For the purpose of squeezing the cells, it is advisable to analyze the cells from the top, as directed in Figure 12A, and the scuttles are available for analysis through the width W of the cells, so more information can be obtained. Therefore, the sperm cells may need to be 11n 也, also 1104 ,, so that the full width w π 卩 width W can be used for analysis before entering the cell assay section 1112. This directional operation provides the most uniform illumination of the cells as they are separated by the device 1104. For most mammalian species, this directed procedure is required to obtain the accuracy required to measure the difference between a sperm-producing female and a male-derived sperm cell. To achieve this goal, the wafer job can include a hydrodynamic focusing section, such as an alignment section (10), to properly position the sperm cells 1104 (see Figure 1, port 13). Section 112 can be used to align the cells from sample injection tube 1132 into sample channel n 122, which is closely related to biological sample 111Q. As illustrated, the segment mo includes the progressive human channels 1124 and 1126 on the same plane, such as a 'plane pupil, and the sheath fluid on the same plane (such as plane X orthogonal to the plane Y). Human passages 1128 and 113〇 are used to properly align and orient the sperm cells flowing through the sample channel iπ2. The sheath fluid entering the secondary channel is organized by a combination of sheath fluids entering the sample channel U22 from each of the secondary channels to align and orient the sperm cells in channel 1122. In the particular embodiment illustrated, the cross-sectional dimensions of the secondary channels 1124 and 1126 are greater than the secondary pass 099122308 28 201105969 lanes 1128 and 1130 such that the amount of sheath fluid that can enter the channel 1122 from these locations is greater than the secondary channel 1128 and The amount of sheath fluid that 1130 entered. Thus, in a particular embodiment not illustrated, the sheath fluid will cause the sperm cells to generally fall on the X plane with their larger width dimension and the smaller thickness dimension generally falls on the gamma plane. In addition, the flow channels 1124, 1126, 1128, and 1130 can also be used to position sperm cells in the center of the channel 112 to better ensure that the cells pass the focus of the optical system that can be part of the cell measurement analysis. The proper orientation is enhanced by the use of a tube 1132 having a beveled end that is aligned with or directed toward the incoming sheath flow from the secondary channels 1124 and 1126. This system creates a laminar flow between the core sample stream of sperm cells exiting the end of the bevel tube 1132 and the sheath fluid passing over the end of the tube 1132. The core sample stream will remain asymmetrical as it passes through the analysis section 1112 of the wafer 11 and the width measured in the X direction is greater than the height measured in the gamma direction. This asymmetry will enable the sperm cells to remain properly oriented as they pass through the analysis section ιι. It will be appreciated that the segment 1120 can be an integral part of the substrate 1102, or can be connected to the base in a suitable manner as would be appreciated by those skilled in the art. In an alternative embodiment, section 112A can be separated from the substrate, with the output of section 1120 being connected to the flow path on substrate 11〇2. Microfluidic Device with Acoustic Coupler: Certain embodiments of the present invention generally relate to a microfluidic device having a phono-coupler positioned in a component of the microfluidic device to destroy the set 099122308 _ 201105969 The cells contained in the pieces. Destruction of the cells allows the release of internal molecular species within the cell so that the substance can be observed or measured by the investigator or medical specialist. After some specific 'in the cell', analysis (such as by flow or imaging cytometry as a non-limiting example), the cells are sorted into containers. Figure μ schematically illustrates a microfluidic device 14A formed on a substrate 1402 on which cells or other substances from a biological sample (not shown) are input to the input port 1410 via cytometry (such as flow Cell cytometry or imaging cytometry) is analyzed in analysis section 1412 (the particular operation performed in analysis section 212 is not critical to the invention). Depending on the results of the analysis performed, the cells are optionally sorted into one or more different wells or chambers 1414. In some embodiments, the sample well 1414 has an outlet port (not shown) in fluid communication therewith to facilitate removal of the sorted sample from the well. The sample fluid can be introduced into the aperture 1414 by appropriate control of the deflector 1415. In one embodiment, the deflector 1415 is a piezoelectric device that can be actuated by an electrical command signal to mechanically direct fluid flow through the flow channel into either of the apertures 1414 depending on the position of the deflector 1415. . In other embodiments, the deflector 1415 is not a piezoelectric device but may be, for example, a bubble that is embedded from the wall to deflect the flow, a deflector that is moved or actuated by the magnetic field, or as is familiar to those skilled in the art. Any other deflector or sorting gate that comes to mind. It should be understood that the various components and sections shown on the substrate 1402 in the illustrated example of FIG. 14 are intended to show the operation of the cell measurement method in a simple schematic form, and that the cell measurement components and sections on the substrate 1402 can be as general. Familiar with the technology 099122308 30 201105969 I think of a big change. Cells can be sorted into different wells or chambers 1414 based on different characteristics of the cells. Cells can be sorted into different chambers 1414 based on the intended future use of the cells. For example, cells having the same characteristics or phenotype can be sorted into one chamber 'fixed in the chamber for review and sorted into another chamber to remain alive in the chamber To perform additional functional measurements, or to be properly stored for use as part of a cell-based therapeutic procedure. In another embodiment, the desired cells can be sorted into an extraction well or chamber, and undesirable cells can be sorted into a waste well or chamber. Alternatively, cells can be deposited into the chamber 1414 based on volume, as opposed to the sorting method. The wafer may include means for physically introducing cells from the analysis section 1412 into the chamber 1414 as is known in the art. Or 'after the analysis is complete, the cells can be caused to leave the substrate 14〇2. For simplicity, the illustration of Figure 14 shows two chambers 1414; however, it should be understood that the microfluidic device can include more or less than two chambers, as will be appreciated by those of ordinary skill in the art. Additionally, chamber M14 is shown as being horizontally aligned proximate to the bottom of substrate 14〇2. However, it will be appreciated that chamber 1414, if present, can be positioned on substrate 1402 as is generally familiar to those skilled in the art. Additionally, for simplicity and ease of illustration, Figure 14 shows a single channel extending between components, regions or sections of substrate 1402. However, it should be understood that a single channel can be a representative of a number of cell measurement channels and a variety of possible channel configurations that are contemplated by those skilled in the art. In accordance with the present invention, at least one component on wafer 1400 includes a sonic energy coupling 099122308 31 201105969 to destroy cells contained in the assembly. As an example, Figure i5 shows a sound coupler (4) in the form of a chamber 1414_probe. Lack of 'should be understood, the sound_coupling can be located in other components of the wafer_,,', including the input hole or other cell collection capacity on the wafer [as illustrated, the acoustic energy coupler (4) 6 is responsive to Sound energy! The received energy vibrates. It will be appreciated that the 'consumer (4) 6 can be sized, shaped, and/or otherwise configured and/or oriented in the chamber 1414 in a manner different from that exemplified, as will be appreciated by those skilled in the art. ... In some specific examples, a sample fluid having suspended cells is collected in a chamber. The Acoustic Auxiliator (4) can be used to vibrate within the sample fluid to destroy the cells in response to the acoustic energy received from the source. The sound energy transferred to the eight (4) 6 can be expressed in the form of sound (four) (such as the amount of material), and the method of tilting and sound can be applied to the sample fluid to give it a lion's cell. In some embodiments, the fitter (10) can be used to provide optimal damage to the cell at its resonant frequency (corresponding to its maximum butterfly vibration or vibration). In some embodiments, cell destruction is sufficiently strong, In order to break up the cellular component' and transfer the (4) molecule of the cell to the sample fluid towel. The internal molecular substance can then be obtained by the researcher or the medical practitioner. In addition, in the illustrated specific example, the cell is Cell measurement analysis and cell sorting are disrupted after chamber (4) 4. However, it should be understood that the light combiner can be placed in different wafer components of the quarantine cells after cell measurement analysis (such as initial fine cell destruction) And the molecular species are released into the initial cell reservoir W1, and the sub-substance can be motivated into the segment 1412 for analysis by the cell. In some embodiments, the acoustic energy source 1420 is separated from the coupler 1416, and Can be combined with an external instrument system for performing cell measurement analysis on a biological sample on a wafer 1400. In other specific examples, an external system that performs cell measurement analysis and self-analysis In addition to the wafer 1400, the acoustic energy source 1420 can be separately applied to the sheet 1400. Microfluidic devices having lenses that produce phase changes: Some specific embodiments of the invention relate generally to microfluidic devices (such as cell measurement wafers) Cell measurement (such as flow cytometry) < Image Cell Measurement) A system for analyzing samples. In order to detect or identify cells during a cell measurement operation, a source of electromagnetic radiation (such as visible light), such as a laser, is directed to the detection zone. When the cells pass through the detection zone within the flow channel, the light source causes the cells to fluoresce. This effect can be enhanced by the addition of certain dyes before the cells reach the 4 test zone, as described above. In some applications, it may be necessary to incorporate various optical components into the substrate of the device or wafer to alter the light scattering properties of the various surfaces. In particular, light shaping optics such as holographic elements or freeform lenses and mirrors can be used to deliver intensity/phase/polarization space patterns, or other light offset effects on various surfaces of the wafer. Because such components can be formed or etched into the wafer substrate, once the initial mold or etch process has been designed, the cost of copying is minimal. More specifically, the lens used to focus the incident excitation beam can also be fabricated to also deliver a particular spatial light pattern within the channel measurement region. 099122308 33 201105969 FIG. 16 schematically shows a specific example in which a focus lens 1602 is mounted on the surface of the substrate 1604 of the microfluidic device 16A. When sample cell 1606 passes through the pre-measured flow channel 16〇8, the excitation pupil 61〇 is focused on sample cell 1606. The surface 1612 of the focusing lens 1602 has been formed, etched, or otherwise processed to impart a spatially varying phase change to the incident excitation light 161. The phase change is converted into an optical holographic effect on the detection area that can be generated to form a particular spatial pattern or design. Surface 1612 may be coated with a UV sensitive material during manufacture of focusing lens 1602. The pulsed laser can then be used to scan the surface 1612 to expose the UV material and produce small changes in the surface depending on the desired pattern or image. When the focusing lens 16〇2 is later mounted on the substrate 1604 and receives the excitation light 1610, it will produce a holographic effect within the detection channel 1608. In some embodiments, the focusing lens 16〇2 can be made entirely of UV-sensitive material, rather than just the surface ι612 made of a UV-sensitive material. In other embodiments, variations in surface 1612 can be achieved by injection molding or other means known in the art for replicating holographic effects. Other types of free-form lenses and mirror shapes can be constructed in a manner similar to holographic elements, by first constructing the original image using micromachining techniques known in the art, such as diamond turning from computer-designed shapes ( Master copy), in combination with focus lens 1602. This artwork can then be replicated using injection molding or other means known in the art. The particular pattern or desired holographic effect can be selected for a variety of uses. By way of a non-limiting example, a holographic lens or free-form light can be used to create a spatially varying pattern to determine the unique characteristics of the detection channel (4) e). · The amount of seams, (iv) Flow channels are often used in parallel in cell measuring devices (such as 'same,,, field cell sorters or microfluidic devices) to measure and / or divide the g & Monthly package. In such systems, the optical optics and associated electronic processing equipment must be able to distinguish signals received from individual channels. This is accomplished by directing the single-excitation source to all channels simultaneously 'but changing or shaping the excitation beam pattern spatially so that different excitation patterns are "to the detection domain of each channel. When the cell passes through the beam pattern of a particular channel, the '-(four) tree „_peak will be generated in the decorated fluorescent signal. The phase change lens discussed above can be used in this case to produce a full & phase effect. The holographic _ effect converts the uniform beam into a unique spatial pattern within the detection area of each channel. Microfluidic device with transmissive mirror and mirror: In the prior art system, one side of the system is applied to the side of the fine-off device, and the measuring optics detects only the fluorescence from the side of the sample cell. Transmitters or other optics are often used to focus the emitted photons partially on the speculating optics, however, this still limits the amount of photons captured from the fluorescent sample cells because the photons emitted in the opposite direction are not Captured by a transmission mirror. The intensity of the excitation light can be increased to produce more photons that are produced by fluorescence, however, this also increases the amount of noise in the signal. Another solution is to use multiple sets of detection optics placed in different locations, but this adds significant expense. In order to capture as much of the photons produced by the fluorescing 099122308 35 201105969 sample cells in the case of a single detection optics, the mirror can be placed on the opposite side of the sample cells from the detection optics. The mirror will capture a large number of photons that are not captured by the mirror, reflect it back through the mirror, and enter the detection optics. This allows the system to capture more emitted photons for a given excitation light intensity without increasing the noise in the received signal and without the cost of multiple detection optics. Figure π schematically illustrates a system for analyzing samples using cytometry. System Π00 can include a microfluidic cell measuring device (shown here in side view) formed on substrate 1702 having a detection flow channel 1704 contained therein. For simplicity and ease of illustration, Figure 17 shows a single channel within substrate 17〇2. However, it should be understood that a single channel can be a representative of a plurality of cell measurement channels and a variety of possible channel configurations that are familiar to those skilled in the art. Various other cell measuring components can also be included on the substrate 17〇' but are not critical to the invention. System 1700 can additionally include a transmission mirror π〇6, a mirror 1707, an excitation source 1710, a dichroic mirror 1712, and a detection optics 1714. Transmissive mirror 1706 and mirror 1707 can be mounted or integrally formed in substrate 1702 as illustrated in Figure 17 depending on the particular application and cost considerations, or can be placed on the outside of substrate 1702. The excitation source π ΐ〇 may comprise a laser or other source of light known in the art. Detecting optics 1714 can comprise various sensitizing means known in the art, such as photomultiplier tubes. In operation, excitation source 1710 directs the beam toward dichroic mirror 1712. The dichroic mirror is configured to reflect the wavelength received by the excitation source 1710 to the detection zone 1716 in the channel 1704 of the detection 099122308 36 201105969. When sample cells 1718 pass through detection zone 1716 and emit fluorescence, the resulting photons are emitted in multiple directions. Some of the photons will be emitted back toward the focusing lens Π06, passing through a dichroic mirror 1712 (which is configured to pass the fluorescent wavelengths) and sensed by the detecting optics 1714. Photons emitted from opposite sides of the wafer 1702 will be captured by the mirror 1707, directed back to the mirror 1706, and focused to the detection optics 1714. The relative distance between the transmission mirror 1706, the detection channel 1704, and the mirror 1707 is such that the light reflected from the mirror 1707 is also properly focused by the transmission mirror 1706 to the detection optics 1714. For example, mirror 1707 may need to be closer to detection channel 1704. When the transmission mirror 1706 and the mirror 1707 are directly mounted on the substrate 1702, the detection channel 1704' as shown in FIG. 17 can be biased to ensure that the transmission mirror and the mirror are opposite to the detection channel π〇4. Implemented with proper placement. Microfluidic device with integrally formed optics: various optical devices, such as lenses, are often placed in the path of the incident and/or exiting beams in the microfluidic device to provide a maximum amount of observed sample cells. Photon transfer and recycling. These optical devices also often need to change the numerical aperture (NA) of the light emitted by the detection zone so that it can be properly received by the fiber optic cable. This is necessary because the emission of light is usually very high, and the fiber cable is usually very low. If the emitted light is not reduced before reaching the fiber optic cable, only a small portion of the transmitted light is successfully transmitted through the fiber optic cable. However, one problem is that various incident and outgoing beams are refracted as they pass through various materials of the beam path 099122308 37 201105969. For example, when a focused excitation beam penetrates from the air into the pure phase of the microfluid (which has a normal plane), the beam will be refracted to some extent, making it more difficult to focus the beam on the intended (four) region within the wafer. To overcome this problem, a focusing lens or other optical unit can be formed in the basic body of the wafer because the air gap between the *focal optical device and the wafer substrate is eliminated' and because the m-subject and the crystal substrate are part of the same material. The portion of the same material is formed and retained, thus eliminating the undesired refraction of the wafer: internal beam. Figure 18 illustrates a system 18 using cell cytometry to analyze a sample. System 1800 can include a microfluidic device The substrate (which is shown in the side view in the figure) has a compensated channel brain contained therein. For simplicity and ease of illustration, ϋ 18 shows the cell measuring a single channel within the wafer cassette. It will be appreciated that the single-channel can be a representative of a plurality of cell measurement channels and sinister channel configurations that are familiar to those skilled in the art. Various other cell measurement components can also be included in The cell measurement wafer 18 〇 2 described herein is not critical to the invention. The substrate 1802 additionally comprises an optical device 18 〇 6 shown here as a focusing lens. The optical device 1806 is integrally formed on the substrate 18 〇 In the substrate of 2. This section can be implemented by various methods known in the art. For example, the optical device 18〇6 can be formed in an injection mold for manufacturing the substrate 1802. In other specific examples, After the basic wafer shape is formed, the optical device 1806 can be processed into the wafer substrate 'or otherwise formed outside the wafer substrate. 099122308 38 201105969 In the known art, the excitation light source 1810 directs the light beam toward the dichroic mirror 1812. Dichroic mirror The 1812 is configured to reflect the wavelength received from the excitation source 181A to the detection region 1814 within the detection channel 1804. Various other optical components known in the art can be used to direct the excitation beam to the optical component 18Q6. The details are not critical to the present invention. When the sample cell 1816 passes through the detection region 1814 and is excited by the excitation beam to emit fluorescence, the resulting photons will be illuminated by the optical device 1 The 806 collects or focuses, passes through a dichroic mirror 1812 (which is configured to pass the fluorescent wavelengths), and is ultimately detected by the detecting optics 1816. The detecting optics 1816 can comprise various sensitencies known in the art. Means, such as photomultipliers. In some embodiments, other components known in the art, such as fiber optic cables, can be used to direct the emitted light beam to the detection optics 1816. It should be understood that the optical device 1806 Any optical component can be included for focusing, propagating, filtering, or otherwise processing the beam of light received by the sample cells 1816 within the assay region 1814. In addition, multiple optical components can be placed on the substrate. In one non-limiting embodiment, additional optics 1818 having a reflective film can be formed in the opposite surface 182 of the substrate 1802 to direct additional photons back to the detection optics 1816. In some deer applications, it is desirable to form an optical device 1818 in the form of an invading (as opposed to a protrusion) in the surface 1820, as illustrated in Figure 18. This allows the surface 1820 of the substrate 18〇2 to be placed flush with the corresponding plane within the mounting device (not shown). Microfluidic devices with non-integral block selective reagent delivery structures: In many cases, reagents may need to be added to activate or otherwise prepare samples prior to performing cell measurement analysis 099122308 39 201105969. The reagents required can be used in the manufacture of fluidic devices, thereby eliminating the need for the user to add at the time of use. This requires many different types of devices to be maintained by the manufacturer = however only applicable: a few applications. In order to increase manufacturing efficiency and reduce inventory: the structure of the present invention can be manufactured with microfluidizers to produce an early microfluidic device that accepts many different reagent delivery structures. Figure 19 (4) The microfluidic device that the fabric of the Weirong sub-contact structure earned. The device measurement may include a cell measurement analyzing portion 19G4 connected to the reagent containing portion 1906 by the fluid flow channel 19 () 8 . The device simplification also includes sample receipt 1910, which is connected to the cell measurement analysis portion 19〇4 by the microfluidic flow channel 19!2. In some embodiments, 'the reagent delivery structure 1902 is mated with the reagent receiving portion leg before the sample is added to the device 19GG. This cooperation can be performed by the user at the time of use or by the manufacturer prior to carrying the device. The sample and reagent mixture contained therein were subjected to cell measurement analysis. For example, the 'cell measurement assay section can sort the desired cells into the extraction well 1914 and sort the undesirable cells into the waste well 1916. The particular operation performed in the cell measurement analysis section 1904, as well as the specific guidance of the microfluidic channel, is not critical to the present invention. The reagent housing portion 1906 is fabricated with a plurality of receiving apertures 1918 configured in a grid or other standardized configuration. The various reagent delivery structures 丨9〇2 can then be fabricated by 099122308 40 201105969 having a reagent portion 1920' when the reagent delivery structure 1902 is mated with the reagent receiving portion 1906, the reagent portions 1920 corresponding to at least one receiving aperture 1918 The location contains reagents. In some embodiments, the location of a particular reagent type in the reagent delivery structure 1902 will also be normalized. For example, any reagent at the position corresponding to a particular receiving aperture 1918 is always of the same type. Because of this, the device used to apply the reagent to the reagent delivery structure 19〇2 can be optimized during manufacture: if a reagent is being applied to a particular reagent delivery structure 19〇2, then a certain reagent type is always applied In the same location. This also allows the cell measurement analysis section 1904 and various microfluidic flow channels 1908 to be designed and optimized based on the known location of the desired reagent. The reagent delivery structure can be made from any material known in the art for use in storing reagents. For example, the reagent structure 1902 can be in the form of a lacing or other wrapable strip ... once applied to the accommodating portion 1906, the reagent structure 19 〇 2 is held in place with the applied adhesive. In some specific examples, the reagent receiving portion may be recessed in the device, and the reagent transfer, the structure 1902 is made of a rigid material, and is fixedly dimensioned to be concealed and attached to the reagent receiving portion. This ensures that the reagent delivery structure is properly placed. The receiving hole 1918 may also include a needle that can pierce the reagent portion and release or expose the reagent contained in the reagent portion 1920 to the receiving hole (9) 8 and the channel 1908. The ’'s member shows another specific example' in which the reagent contained in the reagent structure 2GG2 is held in the structure by the soluble barrier 2_. Activation 099122308 41 201105969 Material 2045 is included in reagent containment portion 2006 within device 2000. Once the reagent structure 2002 is applied to the reagent containing portion 2〇〇6, the activating material 2〇45 causes the soluble barrier rib 2040 to dissolve, thereby releasing the reagent 2020 into the reagent receiving hole 2018 and the flow channel 2〇〇8. In some embodiments, a focused laser beam or sonic energy applied by an external source can be used to cause the soluble barrier 2〇 to dissolve. In other embodiments, the 'soluble barrier 2 〇 4 〇 and/or the activation material 2 〇 45 may be made of a temperature sensitive material, whereby the temperature is raised after the reagent structure 2 〇〇 2 is fitted to the device 2 〇〇〇 'This will cause the soluble barrier 2〇4〇 to dissolve. Various materials that exhibit this temperature sensitive property are known in the art. For example, the reagent can be placed in agarose, which will keep the cells contained in the reagent until the agarose is heated to dissolve. In some embodiments, the soluble barrier 2〇4() can be combined with a micro-motor structure (MEIVISMS° microfluidic device with a light waveguide in the flow channel: some specific examples of the invention are generally Concerning microfluidic devices, such as cell measuring wafers, having cell measurement channels that are grouped to form an optical waveguide to better direct illumination light through the channel. In some embodiments, the cell measurement channel is adapted to be an optical waveguide, Including: selecting a material having a desired refractive index to form a channel wall. In addition, the effect of the optical waveguide channel can also be determined by the refractive index of the fluid passing through the channel. The waveguide channel of the present invention is here for the sake of brevity. Discussed to direct visible light, however, light is only one non-limiting embodiment of the many possible types of electromagnetic radiation that can be directed by the waveguide channel. Figure 21 is a schematic illustration of a system 2100 having an instantiation on a microfluidic device 099122308 42 201105969 The sex cell measurement channel 2102 is part of a cell measurement analysis (such as flow cell measurement or image cell measurement) performed by the device ( The steady cell measurement analysis operation is not critical to the present invention. As illustrated, the channel 21〇2 has an angle 2103 that constitutes two substantially vertical segments 21〇2a and 2102b. The system 2100 follows the channel Section 2102a includes illumination system 2106 and light collection system 2107, and the light system is used to illuminate the cells for partial detection of cellular measurement analysis. In the illustrated embodiment, light collection system 21〇7 is Positioned such that its optical axis is aligned with the axis of the flow through channel section 2102a, and illumination system 2106 is positioned such that its optical axis is orthogonal to the flow axis through passage section 21〇2. It will be appreciated that system 2106 and The particular organization, location, and operation of 2107 must be capable of properly coupling radiation into the system, but is not critical to the present invention, which is a suitable system that is generally contemplated by those skilled in the art. The overall flow through passage 2102 is illustrated by the large arrow 2120 indicated in the passage. The light emitted by system 2106 is internally reflected throughout collection 21 2 to collection system 2107, and is generally indicated by arrow 2 122. Thus, channel 2102 can act as an optical waveguide. Channel 2102 receives illumination from illumination system 2106 and transmits it along the length of channel section 2102a and out toward light collection system 21〇7. To accomplish this, the channel wall 2104 of the channel 2102 can have an appropriate index of refraction along at least the segment 21a 2a such that it internally reflects light as it passes through the segment 2102a. Additionally, the channel wall 2104 is along the segment. All or part of the length of 2102a may be obscured (eg, with a reflective coating) to cause light to be emitted only at the desired location. This 099122308 43 201105969 configuration allows the channel to illuminate only in the desired area, which may be continuous Or multiple points of launch. In some embodiments, the channel 2102 can include a wall section 2108 aligned with the illumination system 21A6 and a wall section 2109 aligned with the light collection system 2107 to allow light to be emitted into and out of the channel. Additionally, in some embodiments, wall sections 2108 and 2109 can be constructed of different materials having different refractive indices than the remainder of wall 2104, the different materials and refractive indices being designed to allow light to be emitted therethrough. In another embodiment where the channel wall 2104 is obscured, the wall sections 2108 and 2109 can remain unobstructed to allow light to pass. A microfluidic device having an optical waveguide and a reflective surface in the flow channel: In some embodiments, the reflective surface can be positioned at one end of the optical waveguide channel to further aid in guiding illumination. Figure 22 is a schematic illustration of a system 2200 having an exemplary cell measurement channel 2202 containing a channel wall 2204 on a microfluidic device as a cellular measurement analysis of the device (such as flow cytometry or imaging cytometry 5 One of the parts (specific cell measurement analysis operations are not critical to the invention). As illustrated, channel 2202 has an angle 2210 between substantially vertical channel sections 2202a and 2202b and an angle 2203 between substantially vertical sections 2202b and 2202c. Additionally, the channel section 2202b integrally includes a first end 2240 adjacent the corner 2210 and a second end 2242 adjacent the angle 2203. System 2200 includes illumination system 2206 and light collection system 2207 along channel section 22〇2b, and the light system is used to illuminate cells, 099122308 44 201105969. In the illustrated embodiment, the liquid Γ Γ 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 22 cross. It will be appreciated that the system and the intermediate readings, (four) and operations must (10) ride properly and in a manner that is not _ for the purposes of the present invention, and such systems are suitable systems as would be apparent to those skilled in the art. In addition, a reflective surface, such as mirror 223, having a mirror face, is positioned adjacent to the first end 2240 that is generally opposite the fiber collection system fiber. As shown in (4), mirror 2 (10) can be combined with the wall of the wall. The mirror may be a spherical shape, or it may have other shapes suitable for reflecting the reflected light coupled by the channel waveguide back down the channel 2202. In other embodiments, the forceps 2230 can be a deformable mirror, which is generally referred to in the art as <Wi# (adaptive Qptie). This recording allows it to be quickly adjusted under the control of the feedback system. The external biological sample passes through the entire flow of the channel 22〇2, as illustrated by the large head 2220 indicated in the channel. Light emitted from system 2206 is internal to the entire channel. The ground is reflected to the collection system 2207 and is generally indicated by arrow 2222. Thus, channel 2202 can act as an optical waveguide. Channel 2202 receives illumination from illumination system 22A6 and transmits it along the length of channel section 2202b and out toward light collection system 22A7. To achieve this, the channel wall 2204 of the channel 2202 can have an appropriate index of refraction at least along the segment 22〇2b, 099122308 45 201105969, such that it internally reflects light as it passes through the segment 2202b. Additionally, all or a portion of the length of the channel wall 2204 along the length of the segment 2202b can be masked (e.g., with a reflective coating) to cause light to be emitted only at the desired location. This configuration allows the channel to illuminate only in the desired area, which can be continuous or multi-point transmission. In addition, the spherical mirror 2232 is positioned to reflect any portion of the light that can reach the mirror 2230 at the end 224, and is reflected back toward the light collecting system 2207 at the end 2242. Using mirror 2230, the efficiency and utility of the optical waveguide channel is improved by preventing the transmission of light from system 2206 to the end 2240 opposite system 2207. In some embodiments, the channel 2202 can include a wall section 2208 aligned with the illumination system 2206 and a wall section 2209 aligned with the light collection system 2207 to allow light to be emitted into and out of the channel. Additionally, in some embodiments, wall segments 2208 and 2209 can be constructed of different materials having a different refractive index than the remainder of wall 2204, which is designed to allow light to be emitted therethrough. In another embodiment where the channel wall 2204 is obscured, the wall sections 2208 and 2209 can remain unobstructed to allow light to pass. Microfluidic Devices for Virus Detection and Sorting: Certain embodiments of the present invention are generally directed to detecting virions in a body sample by means of a microfluidic device such as a cell measuring wafer This is accomplished by performing a cell measurement analysis on the sample and sorting the virions into a well or chamber mounted on the wafer. Cell measurement analysis can be analyzed by flow cell measurement 099122308 46 201105969 or image cell measurement analysis. Sorting virions from body samples allows the researcher or medical professional to capture the virus population for further observation or testing. As used herein, the term "cyt〇metry" is used broadly and is intended to include measuring any suitable substance, including cells, and/or virions, as two non-limiting examples. System 2300 (as schematically illustrated in Figure 23) includes a microfluidic device formed on substrate 23A2 that allows detection of virions during cell measurement analysis, and sorting of virions after analysis. As part of the system 2300, a body sample (not shown) is input to the input cassette 2310, via cytometry analysis on the substrate 23〇2 in the analysis section 2312 (specific operations performed in the analysis section 2312 for For the purposes of the present invention; the result of the analysis performed by the canister of _, input to the input 埠 23U) and the virus in the body sample detected in the analysis section 2312: can be sorted to - or Add more than 4 to the gym. In some embodiments, the sample well 23A has an outlet port (not shown) in fluid communication therewith to facilitate removal of the sorted sample from the well. The sample fluid can be introduced into the aperture 2314 by appropriate control of the deflector. In one of your limbs, the deflector is placed over the wire, and it can be used to change the position of the deflector legs to mechanically (4) = the flow of the moving channel into any hole. 2314. In other embodiments, the piezoelectric device' may be, for example, a self-embedded body such that the _ =: hunter is moved or actuated by a magnetic field, or any other deflector as can be appreciated by me. Or sorting gates. 099122308 201105969 Virions can be further sorted into different wells or chambers 2314 based on their intended future use. For example, virions having the same characteristics can be sorted into chamber 2314a where they are fixed for viewing by electron microscopy and sorted into chamber 2314b in which the chamber is Chemical reactions that were studied for pharmacological efficacy' remaining samples were sorted into chambers 2314c and 2314d according to other criteria. In other embodiments, all of the chambers can contain virions detected in the analysis section 2312. Wafer 23A can include a means for physically introducing virions from analysis section 2312 into chamber 2314 as is known in the art. Alternatively, the material in the body sample can be used to exit wafer 2300 after the analysis is complete. In the illustrated embodiment, there are four illustrated chambers 2314; however, it should be understood that there may be more or less than four chambers as would be apparent to those skilled in the art. The substrate 2302 having a plurality of chambers 2314 can be designed such that a predetermined amount of virions are sorted into each chamber. For ease of illustration, the chambers 2314 are illustrated as being horizontally aligned, although it should be understood that the chambers may otherwise be positioned on the wafer as would be apparent to those skilled in the art. Additionally, for simplicity and ease of illustration, Figure 23 shows a single channel extending between components, regions, or sections of substrate 2302. However, it should be understood that a single channel can be a representative of a plurality of cell measurement channels and a variety of possible channel configurations that are familiar to those skilled in the art. The body sample input at input 埠 2310 can be a blood sample, a urine sample, a tissue sample, a saliva sample, a cell sample, or a combination thereof, and only some non-limiting examples of non-9999308 48 201105969 are listed. In addition to detecting and sorting the presence of any virions in the body sample, the system 2300 can also be grouped to form the number of virions in the measurement sample. In some embodiments, the cell measurement assay in section 2312 is capable of measuring the number of virions in the sample. In other embodiments, the virions are sorted into chamber 2314 on substrate 23〇2 and may be measured or counted after sorting. Some specific embodiments of the present invention are generally directed to detecting (rather than knife-away) virions in a body sample by performing a cell measurement analysis of the sample on a microfluidic device such as a cell measuring wafer. Cell measurement analysis can be flow cell measurement analysis or image cell measurement analysis. The use of such devices to detect virions via cytometry provides increased safety and reduced exposure to researchers or medical professionals due to the documented nature of the tests on microfluidic devices. In addition, the microfluidic device allows cell measurement analysis to be performed at a relatively slow rate so that small virus particles can be detected. In addition, the microfluidic device allows simultaneous detection of cells and virions via a measuring device. As used herein, the term "cell measurement" is used broadly and is intended to include measuring any suitable substance, including cells, and/or virions, as two non-limiting embodiments. In some embodiments, virions can be labeled with a specific fluorescent probe that recognizes a particular sequence of molecules in the RNA. The cell measuring instrument system can detect the presence of virions based on light scattering and classify the particles based on the intensity and wavelength of the fluorescent emissions. In some embodiments, a plurality of fluorescent probes can be used in the same biological sample, and a plurality of viruses can be detected. 099122308 49 201105969 In addition to the presence of any virions in the body sample entered at input 埠 2310, system 230() can also be grouped to form the number of virions in the measurement sample. In some embodiments, the cell measurement analysis in segment 2312 is capable of measuring the number of virions in the sample. In other specific financial terms, virions can be sorted into wells or chambers on substrate 2302 and can be sized or counted after sorting. The number of virions per unit of blood in a specific sample towel relating to AIDS can be measured by cell measurement analysis. Calculating the number of silk particles (also known as viral load) can be an important measure in assessing the health of AIDS patients. If the sample remains on substrate i after analysis in section 2312, the safety of the processed sample is greatly improved 'because' the entire wafer measurement can be discarded as any other contaminated medical device (e.g., such as a used syringe). A microfluidic device that exhibits a color change to indicate use or result: Some specific embodiments of the invention relate generally to a microfluidic device, such as a cell assay! A wafer that is capable of providing a visible indication after cell measurement analysis to indicate the results of the cell measurement analysis. The cell measurement assay can be a flow cytometry assay, or an image cell assay assay as described above. In one exemplary embodiment, if the analysis produces a positive result, the visible indication is the color change caused by the components of the dye flooding device after the analysis is complete. 24 illustrates a system 2400 in which cells from a cell supply source (not shown) are input into an input port 2410 on a substrate 2402 and analyzed in an analysis section 2412 via cytometry (analysis section 2412) The particular operation occurring in the process is not critical to the invention). Based on the results of the analysis performed,

09912230S 50 201105969 細胞可基於細胞之不同特徵選擇地分選至不同腔室2414中。 在某些具體例中’樣品孔2414具有與其流體連通的出口崞(未 圖示）以有助於自孔中移出經分選的樣品。樣品流體可藉由適 當控制導流器2416而導入至孔2414中。 在一具體例中’導流器2416為壓電裝置，其可用電指令信 號致動，以便視導流器2416之位置之不同而以機械方式將通 過流動通道的液流導入任一個孔2414中。在其他具體例中， 導流器2416不為壓電裝置，而可為例如自壁嵌入以使流向偏 轉的氣泡、藉由磁場移動或致動的偏流器、或如一般熟悉本技 藝者可想到的任何其他導流器或分選閘。 細胞可基於細胞之預期的未來用途分選至不同的孔或腔室 2414中。舉例而言，具有相同特徵或表型的細胞可分選至一 個孔中，在該孔中其被固定以供檢視，且分選至另一孔中，而 在δ亥孔中保持其存活狀怨以進行另外的功能測量。或者，細胞 可基於體積沉積至孔或腔室2414中，此與分選方法相反。晶 片可包括如此技藝中已知的以物理方式將細胞自分析區段 2412導入至腔室2414中之手段。或者，在分析完成之後，細 胞可被促使離開基板2402。出於簡單及易於例示起見，圖24 顯示單一通道在基板2402之諸組件、區域、或區段之間延伸。 然而’應瞭解，單-通道可為熟悉本技#者可想到的多個細胞 測量通道及多種可能的通道組構之代表例。 在圖24中例示之特定具體例中，染料可含納於染料貯存器 099122308 51 201105969 2420中，且被儲存以供稱後在裝置上使用。在細胞被分析且 選擇地分選讀’若分析產生由—些狀準陽性結 果，則染料可被允許流出貯存器242〇，且通過晶片24〇2之諸 組件，經由染料顏色而提供陽性結果之可見的指示。在所例示 之特定具體例中，染料可自貯存器_釋放，流至且流過樣 品通道2409,流過分析區段2412，且流入選擇的腔室期中。 在其他具體例中，染料可自貯存器期(其可為不透明，以使 其中之染料為不可見)流至第二非不透明貯存器（未圖示），盆 中，染料在晶片2400外為可見的。然而，應瞭解，染料” 以熟悉本技藝者可想到的許多不同組構或配置存在。針對此 點，染料貯存器2420被顯示為定位成接近晶片頂部；然而， 應瞭解，貯存器可定位於晶片上之其他處所。貯存器242〇内 所含納且用以提供可見的指示的染料，可為熟悉本技藝者可想 到的任何適當經著色染料。在其他具體例中，貯存器内所含 且用以提供可見的指示的物質，為一般熟悉本技藝者可想到的 另一種適當的著色物質。在其他具體例中，設置—個以:染料 貯存器’使顏色改變藉由以下方法而產生：混合兩個貯存器之 内谷物，而在混合時產生顏色改變。貯存器中之材料可為兩種 流體、一種流體及一種非流體、或兩種非流體。 自貯存器2420釋放染料，可由使用者手工操作晶片 而實現。或者’如在此技藝中已知，染_放可在進行細胞測 置分析的裝置或儀器系統期之控制下，諸如藉由開啟維持 099122308 52 201105969 染料於貯存器2420内的閥來實現。貯存器242〇可採取任何適 宜的實體形式，諸如，形成於基板24〇2之表面的孔，其可保 持敞開或可包括-個蓋子，此蓋子係被膠合於適#位置、用喷 合著基板2402的彈性部件扣接在適當位置、藉由自基板24〇2 之表面延伸的導軌而滑動至適當的位置、或一般熟悉本技藝者 可想到的任何其他適宜的手段。以上實施例意欲僅為許多可能 的組構之非限制性實施例。 另外，如一般熟悉本技藝者可想到，本發明涵蓋了提供細胞 測i为析之結果的可見的指示的其他方式，而染料釋放僅為眾 多可能機構之一個非限制性實施例。舉一個替代性實施例而 言，晶片2400可包括併入其中的感測器，其可用於提供細胞 測里分析之結果的可見的指示。舉另一替代性實施例而言，可 見的指示可藉由設置一個機構來獲得，而該機構係將晶片24〇〇 改入者色溶液或染料溶液中，且讓該溶液擴散通過晶片之諸組 件在另—替代性實施例中，基板2402之一個區段塗有感光 材料’且可因電磁刺激引起的化學反應而改變顏色。舉另-替 代陡貝施例而έ，染料可以乾燥形式沉積於晶片2400上，且 由導机至適當位置的鞘液水合。晶片24〇〇可被組構成在出現 、、’田月匕測里刀析之陽性結果時提供可見的指示，或替代性地可被 組構成提供對應於分析之不同可能結果的不同的可見指示。 在正進行細胞測量分析以確定細胞、組織、或其他物質是否 包括危險或有害成分的具體例中，諸如，在AIDS或肝炎測試 099122308 53 201105969 中，提供陽性結果的可見的指示，可向 提供「晶片2400包括危險或有害的病原之T業人貝 式’研究者或醫學專業人員必要時可滅菌、抛棄 式處理晶片2400。與滅菌組合，顏色 ” 5 '、 Μ奸―料日日日片2400 已被成力地去污。另外，在許多細胞·分析環境中，研究者 =學專業人員可能希望在陽性結果後對細胞樣品進行一或 夕:人另外的細胞測量分析’則讀結轉·。陽料果之可 見的指示’使研究者或醫學專業人員在必要時可立即；;行另外 的測試。 在其他具體例中’晶片期可被組構成僅提供晶片已被用 過的可見指示。舉例而言，提供晶片鳩已被用過的明確指 示，可防止細胞樣品之意外污染，該意外污染在第二細胞樣品 々il過被使用過的晶片2400之情況下可能會發生。如一般熟悉 本技藝者可想到’本發明涵蓋了提供微流體裝置已被用過的可 見的指示的其他方式。舉另一實施例而言，分析區段2412及/ 或腔室2414可包括與其耦合的感測器，該等感測器分別提供 了區段2412中之分析完成、或細胞已被收集於腔室2414中的 可見指示。感測器本身可經由顏色改變來提供可見的指示’或 可在晶片2400上指引可見的指示。在又一實施例中’ β曰曰片2400 上之廢料腔室或通道可預先塗有當接觸流體時會顏色改變的 化學物質或染料。以此方式，内部已有流體流動過的任何晶片 將經由顏色改變而被鑑別為已使用過。 099122308 54 201105969 另外，在某些具體例中，晶片2400可被設計成：當整個細 胞測量分析過程中之一或多個步驟已完成時，提供可見的指 不。舉例而言，一或多個細胞樣品輸入件241〇、分析區段 2412、及腔室2414，可包括提供「與該等組件相關的特定步 驟已凡成的可見的指示」之機構。因此，檢視晶片24〇〇的研 究者或醫學專業人員可輕易地確定細胞測量分析之當前狀 態，諸如，細胞樣品已被載入、細胞測量分析完成、或細胞已 被分選。在細胞測量分析之特定階段提供可見的指示的機構， 可為熟悉本技藝者可想到的提供可見料的任何適當機構。 依,，、'本文所揭示之所有具體例，在基板上使用微流體裝置提 供了許多優點，其-為該微流體裝置可作為抛棄式部分被處 理，從而使得新的微流體裝置可用於分選各個新的細胞樣品。 由此極大地簡化了分選設備之處理，且降低了為防止分選階段 之間交叉污麟進行麟備清潔之複雜性，因為，樣品流過的 許^更體被簡單地拋棄。微流體裝置本身亦極適驗在被抛棄 之前滅菌（諸如，藉由Ύ輻射）。 雖然本發明 已在圖式及前述朗巾得轉，示及說明，但 該等圖式及前述說明應視為具例示性而非_性，應瞭解，僅 較佳具體例已被顯示及說明，且在本發明之 精神和範圍内的所 有變化及修改均有意受到保護。 相關申請案之參考引用： 本申請案主張以下申請案之權益:2_年7月削申請之 099122308 55 201105969 美國5品時專利申請案第61/224,533號、2009年7月7日申請 之美國臣品時專利申請案第61/223,417號、2009年7月7曰申 5月之美國臨時專利申請案第61/223,408號、2009年7月8曰 申3青之美國臨時專利申請案第61/223,732號、2009年7月8 曰申清之美國臨時專利申請案第61/223,735號、2〇〇9年7月8 曰申请之美國臨時專利申請案第61/223,736號、2〇〇9年7月8 曰申清之美國臨時專利申請案第61/223,737號、2〇〇9年7月7 曰申晴之美國臨時專利申請案第61/223,416號、2〇〇9年7月7 曰申清之美國臨時專利申請案第61/223,419號、2〇〇9年7月 10曰申請之美國臨時專利申請案第61/224,528號、2〇〇9年7 月7曰申請之美國臨時專利申請案第61/223,42〇號、2〇〇9年7 月7曰申請之美國臨時專利申請案第61/223,421號、2〇〇9年7 月7曰申請之美國臨時專利申請案第61/223,4〇6號、2〇〇9年7 月7曰申請之美國臨時專利申請案第61/223,41〇號、2〇〇9年7 月7曰申請之美國臨時專利申請案第61/223,4〇7號、2〇〇9年7 月7日申請之美國臨時專利申請案第61/223,4〇9號、2〇〇9年7 月8曰申請之美國臨時專利申請案第61/223,734號，所有該等 申凊案均以全文引用的方式併入本文中。 【圖式簡單說明】 圖1為先前技術微流體裴置之立體圖。 圖2為本發明具體例之的微流體裝置之示意側視圖。 圖3為本發明具體例之微流體裝置之示意側視圖。 099122308 56 201105969 圖4為本發明具體例之具有空間改變激發圖型的平行流動 式細胞測量通道之示意圖。 圖5例不來自圖4之圖型的樣品結果榮光信號。 圖6例示具有以二維方式改變的激發圖型的平行流動式細 胞測量通道之示意圖。 圖7為本發明具體例之微流體裝置之示意立體圖。 圖8為本發明具體例之微流體裝置堆疊之示意側視圖。 圖9為圖8之具體例之微流體裝置之示意底面圖。 圖10為本發明具體例之微流體裝置之示意底面圖。 圖11為本發明具體例之微流體裝置之示意立體圖。 圖12A為例示精細胞之示意俯視圖。 圖12B為例示精細胞之示意側視圖。 圖13為本發明具體例之微流體裝置之一區段之示意立體 圖。 圖14為本發明具體例之微流體裝置之示意立體圖。 圖15為具有音能耦合器的微流體裝置之腔室之示意前視 圖。 圖16為本發明具體例之微流體裝置之一區段之示意側視 圖。 圖17為本發明具體例之微流體裝置之一區段之示意側視 圖。 圖18為本發明具體例之微流體裝置之一區段之示意側視 099122308 57 201105969 圖。 圖19為本發明具體例之微流體裝置之示意立體圖。 圖20為本發明具體例之微流體裝置之一部分之示意側剖面 圖。 圖21為本發明具體例之微流體裝置之一部分之示意側剖面 圖。 圖22為本發明具體例之微流體裝置之一部分之示意側剖面 圖。 圖23為本發明具體例之微流體裝置之示意立體圖。 圖24為本發明具體例之微流體裝置之示意立體圖。 【主要元件符號說明】 10 微流體裝置 12 基板 14 流體流動通道 16 埠 18 埠 20 埠 22 流體流動通道 24 樣品注入管 200 (微流體、細胞測量）系統 202 基板，（細胞測置）晶片 204 (偵測）流動通道；（微流體)流動通道 099122308 58 201105969 206 激發光源 208 二向色鏡 210 偵測區 212 (聚焦)透鏡 214 樣品細胞 216 第一末端 218 (多芯）光纖纜線 220 (聚焦)透鏡 222 第二末端 224 第三末端 226 偵測光學器件 300 (微流體、細胞測量）系統 302 (細胞測量）晶片 306 光源 314 樣品細胞 316 末端 318 光纖纜線 322 末端 326 偵測光學器件 402 條狀物 404 細胞 406 偵測區域 099122308 59 201105969 408 (流動）通道 410 (流動)通道 412 (流動)通道 508 螢光信號 510 螢光信號 512 螢光信號 602 (激發）圖型元件 604 細胞 700 (微流體)裝置；（微流體、細胞測量）系統 702 基板；晶片 708 流動通道 709 通道 710 輸入埠 712 分析區段 713 分選通道 714 腔室；（樣品）孔 714a 腔室；（樣品）孔 714b 腔室；（樣品)孔 720 (樣品）孔 720a 底面 722 電磁體；磁體 724 細胞(群） 099122308 60 201105969 730 (輸入)埠 732 (廢料)埠 740 電源線 750 導流器 800 (微流體)裝置；堆疊配置 802 (細胞測置）晶片 802a 頂面；（前)表面 802b 底面；（背）表面 804 腿 805 腳部分 1002 晶片 1004 矩形腳；腿 1100 (微流體)裝置；晶片 1102 基板 1104 (精）細胞 1105 頭 1106 尾 1110 輸入埠；生物樣品 1112 分析區段 1114 腔室；（樣品)孔 1116 導流器 1120 (對齊）區段 099122308 61 201105969 1122 1124 1126 1128 1130 1132 1400 1402 1410 1412 1414 1415 1416 1420 1600 1602 1604 1606 1608 1610 1612 1700 樣品通道 (鞘液進入、鞘流)次通道 (勒液進入、勒流)次通道 (勒液進入、鞘流)次通道 (鞘液進入、鞠流)次通道 樣品注入管；（斜面）管 (微流體)裝置；晶片 基板 輸入璋 分析區段 腔室；（樣品)孔 導流器 (音能)耦合器 音能源；來源 (微流體)裝置 聚焦透鏡 基板 樣品細胞 (偵測)流動通道；（偵測)通道 激發光 (聚焦透鏡)表面 (微流體、細胞測量）系統 099122308 62 201105969 1702 基板，晶片 1704 (偵測)流動通道；（偵測)通道 1706 透射鏡；（聚焦)透鏡 1707 反射鏡 1710 激發光源 1712 二向色鏡 1714 偵測光學器件 1716 /(貞測區 1718 樣品細胞 1800 (微流體、細胞測量）系統 1802 (細胞測量）晶片；（微流體裝置)基板 1804 债測通道 1806 光學器件 1808 (基板)表面 1810 激發光源 1812 二向色鏡 1814 偵測區域 1816 樣品細胞；偵測光學器件 1818 光學器件 1820 (基板)表面 1900 (微流體)裝置 1902 試劑結構；（試劑)傳遞結構 099122308 63 201105969 1904 (細胞測量）分析部分；（細胞測量）分析區段 1906 試劑收容部分 1908 (微流體)流動通道 1910 樣品收容埠 1912 (微流體)流動通道 1914 萃取孔 1916 廢料孔 1918 收容孔 1920 試劑部分 2000 (微流體)裝置 2002 (試劑)結構 2006 試劑收容部分 2008 流動通道 2018 試劑收容孔 2020 試劑 2040 (可溶)障壁 2045 活化材料 2100 (微流體、細胞測量）系統 2102 (細胞測量)通道；（通道）區段 2102a (通道）區段 2102b (通道）區段 2103 角 099122308 64 201105969 2104 通道壁 2106 (發光）系統 2107 (光收集）系統 2108 壁區段 « 2109 壁區段 2120 大頭箭頭；（液流）方向 2122 箭頭；（反射）方向 2200 (微流體、細胞測量）系統 2202 (細胞測量)通道 2202a (通道）區段 2202b (通道）區段 2202c (通道）區段 2203 角 2204 通道壁 2206 (發光）系統 2207 (光收集）系統 2208 壁區段 2209 壁區段 2210 角 2220 大頭箭頭；（液流)方向 2222 箭頭；（反射）方向 2230 鏡子 099122308 65 201105969 2232 (球面）鏡面 2240 (第一）末端 2242 (第二)末端 2300 (微流體、細胞測量）系統；晶片 2302 基板 2310 輸入埠 2312 分析區段 2314 腔室；（樣品)孔 2314a 腔室；（樣品)孔 2314b 腔室；（樣品)孔 2314c 腔室；（樣品）孔 2314d 腔室；（樣品）孔 2316 導流器 2316a 導流器 2316b 導流器 2316c 導流器 2400 (微流體、細胞測量）系統；晶片 2402 基板，晶片 2409 (樣品）通道 2410 (細胞樣品)輸入件；輸入埠 2412 分析區段；（進行細胞測量分析)裝置、儀器系統 2414 腔室；（樣品)孔 099122308 66 201105969 2416 導流器 2420 (染料)貯存器 T (精細胞頭)厚度 W (精細胞頭）寬度；全寬 X 平面 Y 平面 099122308 6709912230S 50 201105969 Cells can be selectively sorted into different chambers 2414 based on different characteristics of the cells. In some embodiments, the sample well 2414 has an outlet port (not shown) in fluid communication therewith to facilitate removal of the sorted sample from the well. The sample fluid can be introduced into the aperture 2414 by appropriately controlling the deflector 2416. In one embodiment, the deflector 2416 is a piezoelectric device that can be actuated by an electrical command signal to mechanically direct fluid flow through the flow channel into either of the apertures 2414 depending on the position of the deflector 2416. . In other embodiments, the deflector 2416 is not a piezoelectric device but may be, for example, a bubble that is embedded from the wall to deflect the flow, a deflector that is moved or actuated by the magnetic field, or as is generally known to those skilled in the art. Any other deflector or sorting gate. Cells can be sorted into different wells or chambers 2414 based on the intended future use of the cells. For example, cells with the same characteristics or phenotype can be sorted into a well where they are fixed for review and sorted into another well while remaining viable in the δHall Complaint for additional functional measurements. Alternatively, cells can be deposited into the well or chamber 2414 based on volume, as opposed to the sorting method. The wafer may comprise means for physically introducing cells from the analysis section 2412 into the chamber 2414 as is known in the art. Alternatively, the cells can be caused to exit the substrate 2402 after the analysis is complete. For simplicity and ease of illustration, FIG. 24 shows a single channel extending between components, regions, or sections of substrate 2402. However, it should be understood that the single-channel can be a representative of a plurality of cell measurement channels and a variety of possible channel configurations that are conceivable in the art. In the particular embodiment illustrated in Figure 24, the dye may be contained in a dye reservoir 099122308 51 201105969 2420 and stored for later use on the device. After the cells are analyzed and selectively sorted to read 'if the analysis produces a positive result, the dye can be allowed to flow out of the reservoir 242 and pass through the components of the wafer 24〇2 to provide a positive result via the dye color. Visible instructions. In the particular embodiment exemplified, the dye can be released from the reservoir, flowed through and through the sample channel 2409, through the analysis section 2412, and into the selected chamber phase. In other embodiments, the dye may flow from the reservoir period (which may be opaque such that the dye therein is invisible) to a second non-opaque reservoir (not shown) in which the dye is visible outside of the wafer 2400 of. However, it should be understood that the dyes are present in many different configurations or configurations as would be appreciated by those skilled in the art. For this, the dye reservoir 2420 is shown positioned close to the top of the wafer; however, it will be appreciated that the reservoir can be positioned at Other locations on the wafer. The dye contained within the reservoir 242 and used to provide a visual indication may be any suitable dyed dye contemplated by those skilled in the art. In other embodiments, the reservoir contains And the substance used to provide a visual indication is another suitable coloring material that is generally known to those skilled in the art. In other embodiments, a dye reservoir is provided to cause a color change to be produced by the following method. Mixing the grains within the two reservoirs and producing a color change upon mixing. The material in the reservoir can be two fluids, one fluid and one non-fluid, or two non-fluid. The dye is released from the reservoir 2420, The user implements the wafer by hand. Or 'as is known in the art, dyeing can be performed on a device or instrument system for performing cell assays. This is accomplished, for example, by opening a valve that maintains 099122308 52 201105969 dye in reservoir 2420. Reservoir 242 can take any suitable physical form, such as a hole formed in the surface of substrate 24〇2, which can be maintained Opened or may include a cover that is glued to the appropriate position, snapped in place with the elastic members sprayed onto the substrate 2402, and slid to the appropriate position by the guide rail extending from the surface of the substrate 24〇2 The position, or generally any other suitable means that may be apparent to those skilled in the art, is intended to be a non-limiting embodiment of many possible configurations. In addition, as is generally understood by those skilled in the art, the present invention encompasses Other ways of providing a visual indication that the cell is the result of the analysis, while dye release is only one non-limiting embodiment of many possible mechanisms. For an alternative embodiment, the wafer 2400 can include a sense of incorporation therein. a detector that can be used to provide a visual indication of the results of the intracellular analysis. For another alternative embodiment, the visible indication can be set by The mechanism obtains, and the mechanism converts the wafer 24 into a color solution or dye solution and diffuses the solution through the components of the wafer. In another alternative embodiment, a section of the substrate 2402 is coated with The photosensitive material 'and may change color due to chemical reactions caused by electromagnetic stimulation. Alternatively, instead of the steep shell application, the dye may be deposited on the wafer 2400 in a dry form and hydrated by a sheath to a suitable position. The 24 〇〇 can be grouped to provide a visual indication of the positive result of the occurrence, the 'Tian Yue 匕 刀 , , , , , or alternatively can be grouped to provide a different visible indication corresponding to the different possible outcomes of the analysis. In a specific example where cell measurement analysis is being performed to determine whether a cell, tissue, or other substance includes a dangerous or harmful component, such as in AIDS or hepatitis test 099122308 53 201105969, a visible indication of a positive result is provided, which may be provided The wafer 2400 includes a hazardous or harmful pathogen T-Bei's researcher or medical professional who can sterilize and discard the wafer 2400 as necessary. In combination with sterilization, the color "5", smuggling - material day 2400 has been strongly decontaminated. In addition, in many cell and analysis environments, researchers = academic professionals may wish to have positive results after the cells The sample is subjected to one or the evening: the additional cell measurement analysis of the person 'reads the carry-over. The visible indication of the positive fruit' enables the researcher or medical professional to immediately if necessary;; performs additional tests. The 'wafer period can be grouped to provide only a visible indication that the wafer has been used. For example, providing a clear indication that the wafer cassette has been used to prevent accidental contamination of the cell sample, the accidental contamination in the second cell sample This may occur in the case of a used wafer 2400. As is generally known to those skilled in the art, the present invention encompasses other ways of providing a visual indication that a microfluidic device has been used. Another embodiment In other words, the analysis section 2412 and/or the chamber 2414 can include sensors coupled thereto that provide analysis in the section 2412, respectively, or the cells have been collected in the chamber 2414. Visible indication. The sensor itself may provide a visible indication via color change' or may indicate a visible indication on the wafer 2400. In yet another embodiment, the waste chamber or channel on the beta sheet 2400 may be pre- Painted with a chemical or dye that changes color when exposed to a fluid. In this way, any wafer that has internal fluid flow will be identified as having been used via color change. 099122308 54 201105969 In addition, in some specific examples In the middle, the wafer 2400 can be designed to provide a visible indication when one or more steps have been completed throughout the cell measurement analysis. For example, one or more cell sample inputs 241, analysis segments 2412, and chamber 2414, may include mechanisms for providing "visible indications that have been made to the particular steps associated with the components." Thus, a reviewer or medical professional reviewing the wafer 24 can readily determine the current state of the cell measurement analysis, such as when the cell sample has been loaded, the cell measurement analysis is complete, or the cells have been sorted. The mechanism that provides a visual indication at a particular stage of the cell measurement analysis can be any suitable mechanism that provides a visible material as would be appreciated by those skilled in the art. According to all of the specific examples disclosed herein, the use of a microfluidic device on a substrate provides a number of advantages - for which the microfluidic device can be treated as a disposable portion, such that a new microfluidic device can be used for Select each new cell sample. This greatly simplifies the processing of the sorting equipment and reduces the complexity of preventing the cross-cleaning between the sorting stages, since the sample flowing through the sample is simply discarded. The microfluidic device itself is also highly suitable for sterilization prior to being discarded (such as by xenon radiation). Although the present invention has been shown and described with reference to the drawings and the foregoing description, the drawings and the foregoing description are to be regarded as illustrative rather than singular. It should be understood that only preferred embodiments have been shown and described All changes and modifications that come within the spirit and scope of the invention are intended to be protected. REFERENCES TO RELATED APPLICATIONS: This application claims the benefit of the following application: 2 _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ US Patent Application No. 61/223,417, July 7, 2009, US Provisional Patent Application No. 61/223,408, May 8, 2009, Shen Sanqing, US Provisional Patent Application No. 61 U.S. Provisional Patent Application No. 61/223,735, July 20, 1989, and U.S. Provisional Patent Application No. 61/223,736, 2, 9 U.S. Provisional Patent Application No. 61/223,737, July 7th, 2009 US Provisional Patent Application No. 61/223,419 of Shen Qing, US Provisional Patent Application No. 61/224,528, filed July 10, 2009 U.S. Provisional Patent Application No. 61/223,42, No. 61/223,421, 2〇〇9, 7 U.S. Provisional Patent Application No. 61/223, No. 4, No. 6 of July 7th, July 7th, 2009, U.S. Provisional Patent Application No. 61/223, 41〇, 2〇〇9, 7 U.S. Provisional Patent Application No. 61/223, No. 4, No. 7 filed on July 7th, and U.S. Provisional Patent Application No. 61/223, No. 4, No. 9 and No. 9 filed on July 7, 2009 U.S. Provisional Patent Application Serial No. 61/223,734, filed on Jan. 8, s. BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 is a perspective view of a prior art microfluidic device. Figure 2 is a schematic side view of a microfluidic device of a specific example of the present invention. Figure 3 is a schematic side view of a microfluidic device of a specific example of the present invention. 099122308 56 201105969 FIG. 4 is a schematic diagram of a parallel flow cell measurement channel having a spatially varying excitation pattern according to a specific example of the present invention. Figure 5 shows a sample glory signal from a sample not shown in Figure 4. Fig. 6 illustrates a schematic diagram of a parallel flow cell measurement channel having an excitation pattern that is changed in two dimensions. Figure 7 is a schematic perspective view of a microfluidic device of a specific example of the present invention. Figure 8 is a schematic side view of a stack of microfluidic devices of a specific example of the present invention. Figure 9 is a schematic bottom plan view of the microfluidic device of the specific example of Figure 8. Figure 10 is a schematic bottom plan view of a microfluidic device of a specific example of the present invention. Figure 11 is a schematic perspective view of a microfluidic device of a specific example of the present invention. Fig. 12A is a schematic plan view illustrating a sperm cell. Fig. 12B is a schematic side view illustrating a sperm cell. Figure 13 is a schematic perspective view of a section of a microfluidic device of a specific example of the present invention. Figure 14 is a schematic perspective view of a microfluidic device of a specific example of the present invention. Figure 15 is a schematic front elevational view of a chamber of a microfluidic device having a sound energy coupler. Figure 16 is a schematic side elevational view of a section of a microfluidic device of a specific example of the present invention. Figure 17 is a schematic side elevational view of a section of a microfluidic device of a specific example of the present invention. Figure 18 is a schematic side view of a section of a microfluidic device of a particular embodiment of the invention 099122308 57 201105969. Figure 19 is a schematic perspective view of a microfluidic device of a specific example of the present invention. Figure 20 is a schematic side cross-sectional view showing a portion of a microfluidic device of a specific example of the present invention. Figure 21 is a schematic side cross-sectional view showing a portion of a microfluidic device of a specific example of the present invention. Figure 22 is a schematic side cross-sectional view showing a portion of a microfluidic device of a specific example of the present invention. Figure 23 is a schematic perspective view of a microfluidic device of a specific example of the present invention. Figure 24 is a schematic perspective view of a microfluidic device of a specific example of the present invention. [Main component symbol description] 10 microfluidic device 12 substrate 14 fluid flow channel 16 埠18 埠20 埠22 fluid flow channel 24 sample injection tube 200 (microfluidic, cell measurement) system 202 substrate, (cell measurement) wafer 204 ( Detect) flow channel; (microfluid) flow channel 099122308 58 201105969 206 excitation light source 208 dichroic mirror 210 detection zone 212 (focus) lens 214 sample cell 216 first end 218 (multicore) fiber optic cable 220 (focus Lens 222 Second End 224 Third End 226 Detection Optics 300 (Microfluidic, Cell Measurement) System 302 (Cell Measurement) Wafer 306 Light Source 314 Sample Cell 316 End 318 Fiber Cable 322 End 326 Detection Optics 402 404 cell 406 detection area 099122308 59 201105969 408 (flow) channel 410 (flow) channel 412 (flow) channel 508 fluorescent signal 510 fluorescent signal 512 fluorescent signal 602 (excitation) pattern element 604 cell 700 (micro Fluid) device; (microfluidic, cell measurement) system 702 substrate; wafer 708 flow channel 709 710 input 埠 712 analysis section 713 sorting channel 714 chamber; (sample) hole 714a chamber; (sample) hole 714b chamber; (sample) hole 720 (sample) hole 720a bottom surface 722 electromagnet; magnet 724 cell ( Group) 099122308 60 201105969 730 (input) 埠 732 (waste) 埠 740 power line 750 deflector 800 (microfluidic) device; stack configuration 802 (cell) wafer 802a top surface; (front) surface 802b bottom surface; Back surface 804 leg 805 foot portion 1002 wafer 1004 rectangular foot; leg 1100 (microfluidic) device; wafer 1102 substrate 1104 (fine) cell 1105 head 1106 tail 1110 input enthalpy; biological sample 1112 analysis section 1114 chamber; ) Hole 1116 deflector 1120 (aligned) section 099122308 61 201105969 1122 1124 1126 1128 1130 1132 1400 1402 1410 1412 1414 1415 1416 1420 1600 1602 1604 1606 1608 1610 1612 1700 Sample channel (sheath fluid entry, sheath flow) secondary channel ( Le fluid inlet, Leliu flow secondary channel (Leys fluid inlet, sheath flow) secondary channel (sheath fluid entry, turbulent flow) secondary channel sample injection tube; (slope) tube (microfluidic) device; wafer Substrate input 璋 analysis section chamber; (sample) hole deflector (sound energy) coupler sound energy; source (microfluidic) device focusing lens substrate sample cell (detection) flow channel; (detection) channel excitation light (focusing lens) surface (microfluidic, cell measurement) system 099122308 62 201105969 1702 substrate, wafer 1704 (detection) flow channel; (detection) channel 1706 transmission mirror; (focus) lens 1707 mirror 1710 excitation source 1712 Chromatic mirror 1714 detection optics 1716 / (measurement zone 1718 sample cell 1800 (microfluidic, cell measurement) system 1802 (cell measurement) wafer; (microfluidic device) substrate 1804 debt measurement channel 1806 optics 1808 (substrate) surface 1810 excitation source 1812 dichroic mirror 1814 detection area 1816 sample cell; detection optics 1818 optics 1820 (substrate) surface 1900 (microfluidic) device 1902 reagent structure; (reagent) transfer structure 099122308 63 201105969 1904 (cell measurement Analytical part; (cell measurement) analysis section 1906 reagent containment part 1908 (microfluidic) flow through Road 1910 sample storage cassette 1912 (microfluidic) flow channel 1914 extraction hole 1916 waste hole 1918 housing hole 1920 reagent portion 2000 (microfluidic) device 2002 (reagent) structure 2006 reagent storage portion 2008 flow channel 2018 reagent receiving hole 2020 reagent 2040 ( Soluble) barrier 2045 activation material 2100 (microfluidic, cell measurement) system 2102 (cell measurement) channel; (channel) section 2102a (channel) section 2102b (channel) section 2103 corner 099122308 64 201105969 2104 channel wall 2106 ( Luminescence) System 2107 (light collection) system 2108 Wall section « 2109 Wall section 2120 Head arrow; (Flow) direction 2122 arrow; (Reflection) direction 2200 (Microfluidics, Cell measurement) System 2202 (Cell measurement) channel 2202a (channel) section 2202b (channel) section 2202c (channel) section 2203 angle 2204 channel wall 2206 (lighting) system 2207 (light collection) system 2208 wall section 2209 wall section 2210 angle 2220 head arrow; ) direction 2222 arrow; (reflection) direction 2230 mirror 099122308 65 201105969 2232 (spherical) mirror 2240 (first) end 2 242 (second) end 2300 (microfluidic, cell measurement) system; wafer 2302 substrate 2310 input 埠 2312 analysis section 2314 chamber; (sample) hole 2314a chamber; (sample) hole 2314b chamber; (sample) hole 2314c chamber; (sample) hole 2314d chamber; (sample) hole 2316 deflector 2316a deflector 2316b deflector 2316c deflector 2400 (microfluidic, cell measurement) system; wafer 2402 substrate, wafer 2409 (sample Channel 2410 (cell sample) input; input 埠2412 analysis section; (for cell measurement analysis) device, instrument system 2414 chamber; (sample) hole 099122308 66 201105969 2416 deflector 2420 (dye) reservoir T ( Sperm cell head) thickness W (sperm cell head) width; full width X plane Y plane 099122308 67

Claims (1)

201105969 七、申請專利範圍： 1.一種微流體裝置系統，包含： 基板， 微流體流動通道，形成於該基板中，其中，該流動通道延伸 通過該基板之-部分，此部分適合於對流人該流親道的細胞 便利地進行細胞測量分析； 第-光收集m用於在第-方向上收集由該等細胞發射 的光，該第一光收集裝置產生第一輪出； 第二光收集裝置，可餘在第二方向上收集由鱗細胞發射 的光，該第二光收集裝置產生第二輸出；及 制光學器件，可用於接收該第—輸出及該第二輸出。 2·如申請專利範圍第1項之微流體裝置系統，其中，該第一 光收集裝置及該第二光收集裝置包含多芯光纖鐵線。以 3·如申請專利範圍第！項之微流體|置系統，其中，該等镇 ，光學II件包含—個選自於由以下組成之群的裝置：光電倍增 管、光電一極體、及突崩光電二極體。 4·如申請專利範圍第1項之微流體農置系統，其中，另外包 含： 第一透鏡’可用於賊第m、於該第—光收集裝置上； 及 第透知了用於將5亥第一光聚焦於該第二光收集裝置上。 5.如申請專利範圍第4項之微流體裝置系統，其中，該第一 099122308 68 201105969 透鏡及該第二透鏡係裝設於該基板上。 6. 如申請專利範圍第4項之微流體裝置系統，其中，該第一 透鏡及該第二透鏡係與該基板整體地形成。 7. 如申請專利範圍第1項之微流體裝置系統，其中，另外包 含： 第一光源，可用於產生第一光輸出；及 第一二向色鏡’可用於將該第一光輸出反射至該等細胞，而 該第一二向色鏡進一步可用於允許由該等細胞在該第一方向 上發射的光通過其間。 8. 如申請專利範圍第7項之微流體裝置系統，其中，另外包 含： 第二光源，可用於產生第二光輸出；及 第二二向色鏡，可用於將該第二光輸出反射至該等細胞’而 該第二二向色鏡進一步可用於允許由該等細胞在該第二方向 上發射的光通過其間。 9. 一種偵測樣品中之粒子的方法，此方法包含以下步驟： a) 設置一個微流體裝置，該微流體裝置包含： 基板；及 微流體流動通道，形成於該基板中，其中，該流動通道延 伸通過該基板之一部分，此部分適合於對流入該流動通道的 細胞便利地進行細胞測量分析； b) 在第一方向上捕獲由該等細胞發射的第一光； 099122308 69 201105969 c) 在第二方向上捕獲由該等細胞發射的第二光； d) 組合在步驟(b)及(c)捕獲的該第一光與該第二光.及 e) 對該組合的第—光與第二光進行細胞測量分析。’ 10. 如申請專利範圍第9項之方法，其中： 步驟⑻包含：用多芯光_線之第—末端捕獲該第一光； 及 步驟⑻包含：用該多芯光纖境線之第二末端捕獲該第二光。 11. 如申請專利範圍第9項之方法’其中，另外包含： f) 自該第一方向將第一來源光照射於該等細胞上；及 g) 自該第二方向將第二來源光照射於該等細胞上。 12. -種偵測樣品中之粒子的方法，此方法包含以下步驟： a) 設置一個微流體裝置，該微流體裝置包含： 基板； 第一彳政體流動通道，形成於該基板中，其中，今第一穿 動通道延伸通過該基板之一部分，此部分適合於對流入該第 一流動通道的第一細胞便利地進行細胞測量分析丨及 第二微流體流動通道，形成於該基板中，其中，該第二流 動通道延伸通過該基板之一部分，此部分適合於對流入該第 二流動通道的第二細胞便利地進行細胞測量分析； b) 產生瞄準於該第一流動通道及該第二流動通道的激發光 束； C)在該激發光束到達該第一流動通道之前，以第一方式在空 099122308 70 201105969 間上改變該激發光束；及 d)在該激發光束到達該第二流動通道之前，以第二方式在空 間上改變該激發光束。 13. —種微流體裝置，包含： 基板； 微流體流動通道，形成於該基板中，其中，該流動通道延伸 通過該基板之一部分’此部分適合於對流入該流動通道的細胞 便利地進行細胞測量分析； 樣品收容孔’形成於該基板之板上，該樣品收容孔係流體式 耦接至該流動通道；及 電磁體’裝設於該基板之板上，而且，當被激發時，可用於 在該樣品收容孔内產生磁場。 14. 一種微流體裝置，包含： 基板； 微流體流動通道，形成於該基板中，其中，該流動通道延伸 通過該基板之一部分，此部分適合於對流入該流動通道的細胞 便利地進行細胞測量分析；及 至少一條腿，位於該基板之表面上，該至少一條腿有助於將 該祕流體裝置堆疊於另一微流體裝置上。 15. —種微流體裴置，包含： 基板， 微流體流動通道’形成於該基板中，其中，該流動通道延伸 099122308 71 201105969 通過該基板之一部分’此部分適合於對流入該流動通道的細胞 便利地進行細胞測量分析；及 至少一個流體動力學對齊結構，流體式耦接至該流動通道， 該至少一個流體動力學對齊結構可用於定向該等細胞，使得大 部分该荨細胞據其最大尺寸來分析。 16. —種微流體裝置，包含： 基板； 微流體流動通道，形成於該基板中，其中，該流動通道延伸 通過該基板之一部分，此部分適合於對流入該流動通道的細胞 便利地進行細胞測量分析；及 孔’裝設於該基板之板上，該孔係與該流動通道流體式耦 接，及 音能耦合器，裝設於該孔内。 17. —種微流體裝置，包含： 基板； 微流體流動通道，形成於該基板中，其中，該流動通道延伸 通過該基板之一部分，此部分適合於對流入該流動通道的細胞 便利地進行細胞測量分析；及 透鏡，形成於該基板之板上，該透鏡可用於在空間上改變通 過其間的光之強度。 18. —種微流體裝置，包含： 基板， 099122308 72 201105969 微流體流動通道，形成於該基板中，其中，該流動通道延伸 通過該基板之一部分，此部分適合於對流入該流動通遒的細胞 便利地進行細胞測量分析； 第一透鏡，形成於該基板之板上，該第一透鏡係裝設於該流 動通道之第一側；及 第二透鏡，形成於該基板之板上，該第二透鏡係裝設於該流 動通道之第二側。 19.一種微流體裝置，包含： 基板，具有第一表面； 微流體流動通道，形成於該基板中，其中，該流動通道延伸 通過該基板之-部分，此部分適合於對流入該流動通道的細胞 便利地進行細胞測量分析；及 該透鏡處於該第 透鏡’在該第一表面形成於該基板之板上， 一表面下方。 20. —種微流體裝置，包含： 基板，具有第一表面； 試劑收容孔，形成於該基板之板上； 試劑結構，具有裝設於其上方的試劑， 八甲，將該試劑結構 施加於該第一表面，使該試劑與該試劑收容孔對齊、 試劑至其中；及 微流體流動通道，形成於該基板中，JL中，兮 5亥流動通道係流 體式耦接至該試劑收容孔。 099122308 73 201105969 21. —種微流體裝置，包含： 基板； 微流體流動通道，形成於該基板中，其中，該流動通道延伸 通過該基板之一部分，此部分適合於對流入該流動通道的細皰 便利地進行細胞測量分析；及 光波導器，形成於該流動通道中。 22. —種微流體裝置’包含： 基板； 微流體流動通道’形成於該基板中’其中’該流動通道延伸 通過該基板之一部分’此部分適合於對流入該流動通道的細& 便利地進行細胞測量分析； 光波導器，形成於該流動通道中；及 反射面，裝設於該流動通道内。 23.—種測定藥理學功效的方法，此方法包含以丁步驟 a)設置一個微流體裝置，該微流體裝置包含： 基板； 微流體流動通道，形成於該基板中， ，、τ，该流動通道延 伸通過該基板之一部分’此部分適合於對流入該流動通道的 病毒粒子便利地進行細胞測量分析；及 孔，形成於該基板之板上； b) 將一物質沉積於該孔中； c) 使該等病毒粒子流入該孔中； 099122308 74 201105969 d) 使該等病毒粒子與該物質反應；及 e) 基於該反應以測定該物質之藥理學功效。 24. —種偵測樣品中之粒子的方法，此方法包含以下步驟： a) 設置一個微流體裝置，該微流體裝置包含： 基板； V 微流體流動通道，形成於該基板中，其中，該流動通道延 伸通過該基板之一部分，此部分適合於對流入該流動通道的 細胞便利地進行細胞測量分析；及 染料貯存器，形成於該基板之板上； b) 將一染料沉積於該孔中； c) 對該等細胞進行細胞測量分析；及 d) 在該細胞測量分析完成之後，使該染料離開該染料貯存 器，且進入該流動通道。 099122308 75201105969 VII. Patent application scope: 1. A microfluidic device system comprising: a substrate, a microfluidic flow channel formed in the substrate, wherein the flow channel extends through a portion of the substrate, the portion being suitable for a convection person Flow-through cells conveniently perform cell measurement analysis; first-light collection m is used to collect light emitted by the cells in a first direction, the first light collection device produces a first round; second light collection device The second light output device generates a second output; and the optical device is configured to receive the first output and the second output. 2. The microfluidic device system of claim 1, wherein the first light collecting device and the second light collecting device comprise a multi-core fiber optic wire. Take 3· as the scope of patent application! The microfluidic system of the present invention, wherein the optical member comprises: a device selected from the group consisting of: a photomultiplier tube, a photodiode, and a collapsing photodiode. 4. The microfluidic farm system of claim 1, wherein the first lens 'is available for the thief m, on the first light collecting device; and the first known to be used for 5 hai The first light is focused on the second light collecting device. 5. The microfluidic device system of claim 4, wherein the first 099122308 68 201105969 lens and the second lens system are mounted on the substrate. 6. The microfluidic device system of claim 4, wherein the first lens and the second lens are integrally formed with the substrate. 7. The microfluidic device system of claim 1, wherein the method further comprises: a first light source operable to generate a first light output; and a first dichroic mirror operative to reflect the first light output to The cells, and the first dichroic mirror are further operable to allow light emitted by the cells in the first direction to pass therethrough. 8. The microfluidic device system of claim 7, wherein the method further comprises: a second light source operable to generate a second light output; and a second dichroic mirror operable to reflect the second light output to The cells are further operable to allow light emitted by the cells in the second direction to pass therethrough. 9. A method of detecting particles in a sample, the method comprising the steps of: a) providing a microfluidic device comprising: a substrate; and a microfluidic flow channel formed in the substrate, wherein the flow The channel extends through a portion of the substrate adapted to facilitate cell measurement analysis of cells flowing into the flow channel; b) capturing first light emitted by the cells in a first direction; 099122308 69 201105969 c) Capture the second light emitted by the cells in a second direction; d) combine the first light and the second light captured in steps (b) and (c) and e) the first light of the combination The second light is subjected to cell measurement analysis. 10. The method of claim 9, wherein: the step (8) comprises: capturing the first light with a first end of the multi-core light ray; and the step (8) comprising: using the second end of the multi-core optical fiber The second light is captured. 11. The method of claim 9, wherein the method further comprises: f) illuminating the first source light from the first direction from the first direction; and g) illuminating the second source light from the second direction On these cells. 12. A method for detecting particles in a sample, the method comprising the steps of: a) providing a microfluidic device comprising: a substrate; a first vapor flow channel formed in the substrate, wherein The first perforating channel extends through a portion of the substrate adapted to facilitate cell measurement analysis of the first cell flowing into the first flow channel and a second microfluidic flow channel formed in the substrate, wherein a second flow channel extending through a portion of the substrate, the portion being adapted to facilitate cell measurement analysis of a second cell flowing into the second flow channel; b) generating an aiming at the first flow channel and the second flow An excitation beam of the channel; C) changing the excitation beam between spaces 099122308 70 201105969 in a first manner before the excitation beam reaches the first flow channel; and d) before the excitation beam reaches the second flow channel, The excitation beam is spatially altered in a second manner. 13. A microfluidic device comprising: a substrate; a microfluidic flow channel formed in the substrate, wherein the flow channel extends through a portion of the substrate 'this portion is adapted to conveniently perform cells on cells flowing into the flow channel Measuring and analyzing; a sample receiving hole ' is formed on the board of the substrate, the sample receiving hole is fluidly coupled to the flow channel; and the electromagnet is mounted on the board of the substrate, and when activated, is available A magnetic field is generated in the sample receiving hole. 14. A microfluidic device comprising: a substrate; a microfluidic flow channel formed in the substrate, wherein the flow channel extends through a portion of the substrate, the portion being adapted to facilitate cell measurement of cells flowing into the flow channel An analysis; and at least one leg on the surface of the substrate, the at least one leg facilitating stacking the secret fluid device on another microfluidic device. 15. A microfluidic device comprising: a substrate, a microfluidic flow channel formed in the substrate, wherein the flow channel extends 099122308 71 201105969 through a portion of the substrate 'this portion is adapted to cells flowing into the flow channel Conveniently performing cell measurement analysis; and at least one fluid dynamic alignment structure fluidly coupled to the flow channel, the at least one hydrodynamic alignment structure being operable to orient the cells such that a majority of the cells are according to their maximum size To analyze. 16. A microfluidic device comprising: a substrate; a microfluidic flow channel formed in the substrate, wherein the flow channel extends through a portion of the substrate, the portion being adapted to conveniently perform cells on cells flowing into the flow channel The measurement is performed; and the hole is mounted on the board of the substrate, the hole is fluidly coupled to the flow channel, and the sound energy coupler is disposed in the hole. 17. A microfluidic device comprising: a substrate; a microfluidic flow channel formed in the substrate, wherein the flow channel extends through a portion of the substrate, the portion being adapted to conveniently perform cells on cells flowing into the flow channel Measurement analysis; and a lens formed on the substrate of the substrate, the lens being operable to spatially vary the intensity of light passing therethrough. 18. A microfluidic device comprising: a substrate, 099122308 72 201105969 a microfluidic flow channel formed in the substrate, wherein the flow channel extends through a portion of the substrate, the portion being adapted to flow into the flow through cell Conveniently performing cell measurement analysis; a first lens is formed on the substrate of the substrate, the first lens is mounted on the first side of the flow channel; and a second lens is formed on the substrate of the substrate, the first The two lens system is mounted on the second side of the flow channel. 19. A microfluidic device comprising: a substrate having a first surface; a microfluidic flow channel formed in the substrate, wherein the flow channel extends through a portion of the substrate, the portion being adapted to flow into the flow channel The cells are conveniently subjected to cell measurement analysis; and the lens is on the first lens formed on the substrate of the substrate, below a surface. 20. A microfluidic device comprising: a substrate having a first surface; a reagent receiving hole formed on the substrate; a reagent structure having a reagent mounted thereon, a octa, applying the reagent structure to The first surface is such that the reagent is aligned with the reagent receiving hole, and the reagent is therein; and the microfluidic flow channel is formed in the substrate. In the JL, the flow channel is fluidly coupled to the reagent receiving hole. 099122308 73 201105969 21. A microfluidic device comprising: a substrate; a microfluidic flow channel formed in the substrate, wherein the flow channel extends through a portion of the substrate, the portion being adapted for blisters flowing into the flow channel Conveniently performing cell measurement analysis; and an optical waveguide formed in the flow channel. 22. A microfluidic device comprising: a substrate; a microfluidic flow channel formed in the substrate 'where the flow channel extends through a portion of the substrate' is adapted to facilitate the flow into the flow channel and conveniently Performing cell measurement analysis; an optical waveguide formed in the flow channel; and a reflective surface disposed in the flow channel. 23. A method of determining pharmacological efficacy, the method comprising the step of a) providing a microfluidic device comprising: a substrate; a microfluidic flow channel formed in the substrate, , τ, the flow Channel extending through a portion of the substrate 'this portion is adapted to facilitate cell measurement analysis of virions flowing into the flow channel; and pores formed on the substrate; b) depositing a substance in the well; c Passing the virions into the well; 099122308 74 201105969 d) reacting the virions with the substance; and e) determining the pharmacological efficacy of the substance based on the reaction. 24. A method of detecting particles in a sample, the method comprising the steps of: a) providing a microfluidic device comprising: a substrate; a V microfluidic flow channel formed in the substrate, wherein The flow channel extends through a portion of the substrate adapted to facilitate cell measurement analysis of cells flowing into the flow channel; and a dye reservoir formed on the substrate; b) depositing a dye in the well c) performing cell measurement analysis on the cells; and d) after the cell measurement analysis is completed, leaving the dye exiting the dye reservoir and entering the flow channel. 099122308 75
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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
TWI720129B (en) * 2016-02-17 2021-03-01 奧地利商Ev集團E塔那有限公司 Metrology device and metrology method
CN114231401A (en) * 2022-02-21 2022-03-25 北京芯迈微生物技术有限公司 Nucleic acid detection kit and detection method

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
TWI720129B (en) * 2016-02-17 2021-03-01 奧地利商Ev集團E塔那有限公司 Metrology device and metrology method
CN114231401A (en) * 2022-02-21 2022-03-25 北京芯迈微生物技术有限公司 Nucleic acid detection kit and detection method
CN114231401B (en) * 2022-02-21 2022-05-03 北京芯迈微生物技术有限公司 Nucleic acid detection kit and detection method

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