Sanders J C, Huang Z, Landers J P
Department of Chemistry, University of Virginia, Charlottesville, VA 22904, USA.
Lab Chip. 2001 Dec;1(2):167-72. doi: 10.1039/b107835f. Epub 2001 Nov 28.
This paper describes the development of a technique amenable to the separation of proteins on a microchip by isoelectric focusing (IEF) with entire channel scanning laser-induced fluorescence detection using acousto-optical deflection (AOD). The ability to use AOD to scan the portions of or the entire length of an IEF separation channel allows for high-speed analysis since the mobilization step is circumvented with this technique. Employing no moving parts eliminates mechanical noise and, not only is there no loss of resolution, AOD scanning can potentially increase resolution. The ability of AOD to provide ultra-fast scanning rates (kHz timescale) allows for real-time imaging of the focusing process. This is demonstrated with the separation of naturally fluorescent proteins using entire channel (total scanning range of 2.4 cm) AOD-mediated scanning laser-induced fluorescence detection.
本文描述了一种技术的发展,该技术适用于在微芯片上通过等电聚焦(IEF)分离蛋白质,并使用声光偏转(AOD)进行全通道扫描激光诱导荧光检测。使用AOD扫描IEF分离通道的部分或整个长度的能力实现了高速分析,因为该技术规避了流动步骤。不使用移动部件消除了机械噪声,并且不仅没有分辨率损失,AOD扫描还可能提高分辨率。AOD提供超快扫描速率(千赫兹时间尺度)的能力允许对聚焦过程进行实时成像。使用全通道(总扫描范围为2.4厘米)AOD介导的扫描激光诱导荧光检测对天然荧光蛋白进行分离,证明了这一点。