Schulze Philipp, Belder Detlev
Institute of Analytical Chemistry, Universitat Leipzig, Johannisallee 29, 04103, Leipzig, Germany.
Anal Bioanal Chem. 2009 Jan;393(2):515-25. doi: 10.1007/s00216-008-2452-7. Epub 2008 Nov 1.
Herein, we summarize the current status of native fluorescence detection in microchannel electrophoresis, with a strong focus on chip-based systems. Fluorescence detection is a powerful technique with unsurpassed sensitivity down to the single-molecule level. Accordingly fluorescence detection is attractive in combination with miniaturised separation techniques. A drawback is, however, the need to derivatize most analytes prior to analysis. This can often be circumvented by utilising excitation light in the UV spectral range in order to excite intrinsic fluorescence. As sensitive absorbance detection is challenging in chip-based systems, deep-UV fluorescence detection is currently one of the most general optical detection techniques in microchip electrophoresis, which is especially attractive for the detection of unlabelled proteins. This review gives an overview of research on native fluorescence detection in capillary (CE) and microchip electrophoresis (MCE) between 1998 and 2008. It discusses material aspects of native fluorescence detection and the instrumentation used, with particular focus on the detector design. Newer developments, featured techniques, and their prospects in the future are also included. In the last section, applications in bioanalysis, drug determination, and environmental analysis are reviewed with regard to limits of detection.
在此,我们总结了微通道电泳中天然荧光检测的现状,重点关注基于芯片的系统。荧光检测是一种强大的技术,具有无与伦比的灵敏度,可低至单分子水平。因此,荧光检测与小型化分离技术相结合很有吸引力。然而,一个缺点是在分析之前需要对大多数分析物进行衍生化。这通常可以通过利用紫外光谱范围内的激发光来激发固有荧光来规避。由于基于芯片的系统中灵敏的吸光度检测具有挑战性,深紫外荧光检测目前是微芯片电泳中最常用的光学检测技术之一,这对于未标记蛋白质的检测尤其有吸引力。本综述概述了1998年至2008年间毛细管电泳(CE)和微芯片电泳(MCE)中天然荧光检测的研究。它讨论了天然荧光检测的材料方面和所使用的仪器,特别关注检测器设计。还包括了最新进展、特色技术及其未来前景。在最后一节中,针对生物分析、药物测定和环境分析中的应用,对检测限进行了综述。