Department of Chemistry, University of North Carolina, Chapel Hill, NC, USA.
Electrophoresis. 2010 Aug;31(15):2558-65. doi: 10.1002/elps.201000054.
A capillary-based microelectrophoresis platform for fast serial analysis of single cells is described. In this system, the capillary remains fixed and a two-channel flow system is used to rapidly switch the buffer surrounding the capillary inlet from a physiological buffer to an electrophoretic buffer. Single cells are retained in the physiologic buffer channel utilizing an array of cell microwells patterned into the channel floor. The defined addresses of the cells on the array enable the sequential delivery of individual cells to the inlet of the capillary, where a focused laser pulse lyses the cell. The cell chamber is moved along a preordained route so that the inlet of the capillary is located in the electrophoresis buffer for separation and the physiological buffer during cell sampling. The throughput of the current system is limited by peak overlap between successive samples. Key characterizations of this system including the fluid flow rates, the cell array dimensions, and laser energies were performed. To demonstrate this system, 28 cells loaded with Oregon green and fluorescein were serially analyzed in under 16 min, a rate of 1.8 cells/min.
一种用于快速连续分析单细胞的基于毛细管的微电泳平台被描述。在这个系统中,毛细管保持固定,采用双通道流系统快速切换毛细管入口周围的缓冲液,从生理缓冲液切换到电泳缓冲液。单细胞保留在生理缓冲液通道中,利用微通道地板上的细胞微井阵列。细胞阵列上的定义地址可以使单个细胞顺序递送到毛细管入口,在那里聚焦激光脉冲裂解细胞。细胞室沿着预定路线移动,使得毛细管的入口位于电泳缓冲液中进行分离,而在细胞采样期间位于生理缓冲液中。当前系统的吞吐量受到连续样品之间峰重叠的限制。对该系统的关键特性,包括流体流速、细胞阵列尺寸和激光能量进行了表征。为了演示该系统,28 个装载有 Oregon Green 和荧光素的细胞在不到 16 分钟的时间内被连续分析,分析速度为 1.8 个细胞/分钟。