Department of Chemistry, Chapman Hall Room 251, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599-3216, USA.
Anal Chem. 2010 Feb 1;82(3):967-73. doi: 10.1021/ac902218y.
A microfabricated fluidic device was developed for the automated real-time analysis of individual cells using capillary electrophoresis (CE) and electrospray ionization-mass spectrometry (ESI-MS). The microfluidic structure incorporates a means for rapid lysis of single cells within a free solution electrophoresis channel, where cellular constituents were separated, and an integrated electrospray emitter for ionization of separated components. The eluent was characterized using mass spectrometry. Human erythrocytes were used as a model system for this study. In this monolithically integrated device, cell lysis occurs at a channel intersection using a combination of rapid buffer exchange and an increase in electric field strength. An electroosmotic pump is incorporated at the end of the electrophoretic separation channel to direct eluent to the integrated electrospray emitter. The dissociated heme group and the alpha and beta subunits of hemoglobin from individual erythrocytes were detected as cells continuously flowed through the device. The average analysis throughput was approximately 12 cells per minute, demonstrating the potential of this method for high-throughput single cell analysis.
研制了一种微流控装置，用于使用毛细管电泳（CE）和电喷雾电离-质谱（ESI-MS）对单个细胞进行自动实时分析。微流体结构包含一种在自由溶液电泳通道内快速裂解单个细胞的手段，在该通道中细胞成分被分离，并集成了用于分离成分离子化的电喷雾发射器。洗脱液使用质谱进行了表征。本研究以人红细胞作为模型系统。在这个整体集成的装置中，细胞裂解发生在通道交叉处，使用快速缓冲液交换和电场强度增加的组合。在电泳分离通道的末端并入电动泵，将洗脱液引导至集成的电喷雾发射器。从单个红细胞中分离的血红素基团和血红蛋白的 alpha 和 beta 亚基作为细胞连续流过装置被检测到。平均分析通量约为每分钟 12 个细胞，表明该方法具有高通量单细胞分析的潜力。
