CN100520341C - 液滴操作装置 - Google Patents

液滴操作装置 Download PDF

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CN100520341C
CN100520341C CNB038201623A CN03820162A CN100520341C CN 100520341 C CN100520341 C CN 100520341C CN B038201623 A CNB038201623 A CN B038201623A CN 03820162 A CN03820162 A CN 03820162A CN 100520341 C CN100520341 C CN 100520341C
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安田贤二
一木隆范
冈野和宣
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Yasuda Kenji
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Abstract

液滴操作装置，其包括具有光透过性，实施了防水处理的基板；向该基板上供给液滴的装置；搬运防水基板上的液滴的装置；具有测定液滴状态的装置的系统，例如，包括表面实施了防水处理并具有光透过性的基板11，将试剂或试料的液滴51导入基板11上的溶液送入口21、22、23，光学测定单元61、62、63，外加电场单元71、72、73等，可以显微镜观察液滴的内部，使液滴静止或使其沿任意的方向移动，使液滴与其他液滴反应，对其反应结果测定、分离。

Description

液滴操作装置
技术领域
本发明申请涉及用于操作液滴的新型装置。
背景技术
将培养液中的特定细胞分离、回收在生物学、医学分析上是重要的技术。通常，当因细胞的比重不同而分离细胞时，可以采用速度沉降法进行分离。但是，当如辨别未过敏反应细胞和过敏反应细胞这样，细胞几乎无差异时，必须以用荧光抗体染色的信息或目视的信息为基础将细胞一个个地分离。对于该技术，例如包括细胞分类器(cellsorter)。细胞分类器是通过将荧光染色处理后的细胞以1细胞单位分离滴入具有电荷的液滴中，以该液滴中的细胞有无荧光、光散射量的大小为基础，在液滴下落的过程中在下落方向的法平面方向上以任意方向外加高电场，从而控制液滴的下落方向，分离到置于下部的多个容器中进行回收的技术。对于该技术，Kamarck，M.E.在MethodsEnzymol.第151卷第150页～165页(1987年)有详细的报告。
发明内容
上述以往使用细胞分类器的细胞分离技术使目前为止不可能的以液滴单位的光学信息为基础的观测成为可能，但对自由下落的液滴的内部状态进行光学上显微观察仍是困难的。特别是没有直接确认是否液滴中实际上只含有一个细胞的方法，而且只能在下落的液滴的法平面方向上作用电场，因此基本上只能调节液体下落的角度。此外，为了使高速下落的液滴在短时间内偏转足够的角度，必须作用2000V以上的高电场。此外，采用以往的细胞分类器计测时，计测只是1次，不能使反应液与分离的液滴混合，进而根据其结果进行分离。
因此，本发明申请的课题在于消除以上所述的以往技术的问题，提供新型的液滴操作装置，该液滴操作装置使对含有细胞或微生物的液滴的内部进行观察成为可能，而且使为此而使上述液滴静止，或使其在任意的方向上移动，对同一液滴进行多次光学计测，使液滴与其他液滴反应并计测其反应结果并进行分离等成为可能。
本发明申请为了解决上述课题，提供如下液滴操作装置，其具有在表面实施了防水处理的绝缘性基板上配置多个电极，向该基板上供给液滴的装置、通过向上述多个电极外加直流或交流电场而将防水基板上的液滴搬运的装置、测定液滴状态的装置。
此外，本发明申请提供如下液滴操作装置，其具有使用了显微镜所成光学像或光传感器等测定液滴的pH、离子浓度、吸光度、荧光等的装置作为测定上述液滴状态的装置，此外，还提供具有使多个液滴混合的装置的液滴操作装置。
附图说明
图1为表示本发明申请的基本构成的模式图。
图2为表示本发明申请的实施例的模式图。
图中的符号表示如下内容。
1：绝缘基板
2、3：电极
4：绝缘层
5：液滴
6：物镜
11：基板
12、13、14、15、16：沟
21、22、23：溶液送入口
31、32、33：溶液排出口
41、42、43、44：电子电路
51：液滴
61、62、63：光学测定单元
71、72、73：外加电场单元
81、82、83：液滴导管
91、92、93：脚
具体实施方式
本发明申请具有如上所述的特征，以下对其实施方式进行说明。
首先，使用图1对本发明申请的液滴操作装置的基本构成进行说明。在该例示中，以覆盖配置于绝缘基板1上的多个电极2、3的形式配置绝缘层4。绝缘层表面实施防水处理，对于此时的防水处理可以适当，例如可以考虑采用硅系树脂或氟系树脂涂布的方法、或在基板表面上形成具有不规则碎片形(フラクタル)结构的凹凸面从而赋予防水性的方法等各种方法。
此外，含有细胞或微生物的液滴5以球状液滴的状态保持在基板上。与细胞分类器相同，当向液滴外加电荷时，通过向电极2、3外加直流电场可以控制液滴的移动方向。此外，在没有外加电荷的液滴的情况下，例如，通过在电极2、3之间外加交流电场，可以使细胞或微生物移动。此外，在对液滴中的细胞进行观察时，通过物镜6可以用位相差显微镜像或荧光显微镜像直接对液滴内的细胞或微生物的状态进行观察。为了光学测定，防水处理的基板可以具有光透过性，对于电极也可以同样为透光性的电极。
以下根据图2所示的模式图对实施例的计测例进行说明。
在该实施例所示的装置中，其具备表面实施了防水处理的基板11，将试剂或试料的液滴导入基板上的溶液插入口21、22、23，搬运液体时粗略控制液体行进方向的沟12、13、14、15、16，将计测、分离后的液滴排出的溶液排出口31、32、33，控制基板上的元件的电子电路41、42、43、44，光学测定液滴的状态的光学测定单元61、62、63，控制液滴51的移动方向的外加电场单元71、72、73，防止液滴逾越的液滴导管81、82、83和基板的脚91、92、93。因此，例如，从溶液插入口21导入的液滴51边沿沟12顺着基板11的缓缓的倾斜滚动，边导入第1光学测定单元，根据测定的液滴的状态，在与沟13的交叉点通过外加电场单元71外加电场，使液滴产生分歧。当没有外加电场时，液滴进入下面的分支路14，当外加电场时，沿沟13进入溶液排出口31。当以上的液滴51移动时，也可以使基板11不倾斜，也可以通过调整沟12的宽度和深度控制移动速度。当然可以配置电极对该速度进行控制。
此外，例如，进行到分支路的沟14的液滴重新通过外加电场单元72、73而向沟15或沟16行进。然后，向沟15或沟16行进的液滴由于外加电场单元72或73而一边保持在该场中，一边与从溶液送入口22或23导入的液滴碰撞而融合。然后，在经过必要的反应时间后，液滴再次沿沟14或15滚动，用光学测定单元72、73测定后，在溶液排出口32、33处被回收。在该实施例中，将外加电场单元配置在基板上，但也可以如图1所示那样配置在防水基板表面的下面。此外，可以在溶液送入口21、22、23各处，导入外加了正电荷、负电荷的液滴，也可以导入不带电荷的液滴。此外，在该实施例中，在将多个液滴混合后，只进行光学测定，可以以光学测定的结果为基础对液滴进一步分离。
当然并不限于这些，在本发明申请中，对于其细节可以有各种各样的方式。
如以上详述的那样，根据本发明申请，可以以含有细胞或微生物、甚至化学物质等的微量液体为液滴进行操作、搬运、混合、观察、测定。

Claims (6)

1.液滴操作装置，其具有：表面具有实施了防水处理的绝缘层的基板、配置在该基板和该绝缘层之间的液滴移动或停止操作用的多个电极装置、向基板表面上供给液滴的装置、在基板表面上通过向上述电极装置外加电场而将液滴搬运的装置、测定液滴状态的装置。
2.权利要求1所述的液滴操作装置，其特征在于：具有用光学显微镜测定液滴的内部状态的装置。
3.权利要求1所述的液滴操作装置，其特征在于：具有测定液滴的吸光度的装置。
4.权利要求1所述的液滴操作装置，其特征在于：具有测定液滴的荧光强度的装置。
5.权利要求1所述的液滴操作装置，其特征在于：具有将多个液滴混合的装置。
6.权利要求1～5的任一项所述的液滴操作装置，其特征在于：液滴含有细胞或微生物。
CNB038201623A 2002-08-26 2003-08-26 液滴操作装置 Expired - Fee Related CN100520341C (zh)

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CN109908987B (zh) * 2019-03-20 2021-01-29 山东大学 基于热释电效应无损失移取微量液滴的移液器的制备方法
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