Characterization of cell lysis events on a microfluidic device for high-throughput single cell analysis.
Characterization of cell lysis events on a microfluidic device for high-throughput single cell analysis.
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DOI：
10.1002/elps.201100229
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发表时间：
2011-11
期刊：
影响因子：
2.9
通讯作者：
Ramsey, John Michael
中科院分区：
文献类型：
--
作者：
Hargis, Amy D.;Alarie, Jean Pierre;Ramsey, John Michael
进一步表征了一种能够使用电子细胞裂解以高通量方式快速分析细胞的微流控装置。在进行的实验中，使用具有高帧速率(>；100fps)数据采集的EMCCD相机来研究细胞裂解事件。结果发现，在这种微流控设计中，细胞通过电场的路径会影响注入分析通道的裂解物的量。通过在分析通道上涂覆多胺化合物来逆转电渗流(EOF)，实现了通过电场的可变流动路径的消除。EOF的反转迫使细胞在电场中采取相同的路径。改进的细胞轨迹控制将减少设备对分析施加的偏差，并最大限度地增加每个细胞注入分析通道的裂解物数量，从而提高分析物检测能力。
A microfluidic device capable of rapidly analyzing cells in a high-throughput fashion using electrical cell lysis is further characterized. In the experiments performed, cell lysis events were studied using an EMCCD camera with high frame rate (> 100 fps) data collection. It was found that, with this microfluidic design, the path that a cell follows through the electric field affects the amount of lysate injected into the analysis channel. Elimination of variable flow paths through the electric field was achieved by coating the analysis channel with a polyamine compound to reverse the electroosmotic flow (EOF). EOF reversal forced the cells to take the same path through the electric field. The improved control of the cell trajectory will reduce device-imposed bias on the analysis and maximizes the amount of lysate injected into the analysis channel for each cell, resulting in improved analyte detection capabilities.
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