Fast-lysis cell traps for chemical cytometry
Fast-lysis cell traps for chemical cytometry
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DOI：
10.1039/b719301g
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发表时间：
2008-01-01
期刊：
影响因子：
6.1
通讯作者：
Allbritton, Nancy L.
中科院分区：
文献类型：
--
作者：
Marc, Paul J.;Sims, Christopher E.;Allbritton, Nancy L.
电寻址细胞捕捉器与毛细管电泳联用来分析单个贴壁细胞的含量。用1002F光刻胶在玻璃表面上刻画了由氧化铟锡组成的电极，形成了表面细胞陷阱。通过对电池下方的电极和放置在电池陷阱上方的水介质中的接地电极施加短暂的电场(10ms)，可以在不到66ms的时间内裂解被困在孔洞中的单个电池。在细胞裂解过程中形成的气体仍然局限在1002F光刻胶形成的空腔内。气体在细胞捕捉器中的滞留使细胞捕捉器能够连接到覆盖的毛细管而不会堵塞毛细管。在陷阱中培养的单个细胞被负载荧光素和俄勒冈绿，然后电裂解。通过同时向毛细管施加电场，细胞的内容物被加载到毛细管中并进行电泳分离。在不到两分钟的时间内，奥金绿和荧光素从单个细胞中完全分解。在1002F电池捕捉器下使用单个图案化电极产生了一个简单、易于制造的设计，该设计在浸泡在水溶液中时是坚固的。此外，该设计可以很容易地放大，以创建附着细胞阵列，用于使用单个毛细管进行连续分析或通过与毛细管阵列配对进行并行分析。提高单个贴壁细胞的分析速度将使人们能够更好地了解细胞生理学。
Electrically addressable cell traps were integrated with capillary electrophoresis for the analysis of the contents of single adherent cells. Electrodes composed of indium tin oxide were patterned on a glass surface followed by formation of topographical cell traps using 1002F photoresist. Single cells trapped in the holes could be lysed in less than 66 ms by applying a brief electric field (10 ms) across the electrode beneath the cell and the ground electrode placed in the aqueous media above the cell traps. The gas formed during cell lysis remained localized within the cavity formed by the 1002F photoresist. The retention of the gas in the cell trap enabled the cell traps to be coupled to an overlying capillary without blockage of the capillary. Single cells cultured in the traps were loaded with fluorescein and Oregon Green and then electrically lysed. By simultaneous application of an electric field to the capillary, the cell's contents were loaded into the capillary and electrophoretically separated. Orgeon Green and fluorescein from a single cell were fully resolved in less than two minutes. The use of a single patterned electrode beneath the 1002F cell trap yielded a simple easily fabricated design that was robust when immersed in aqueous solutions. Moreover, the design can easily be scaled up to create arrays of adherent cells for serial analyses using a single capillary or for parallel analysis by mating to an array of capillaries. Enhancing the rate of analysis of single adherent cells would enable a greater understanding of cellular physiology.