Dual-color total internal reflection fluorescence cross-correlation spectroscopy
Dual-color total internal reflection fluorescence cross-correlation spectroscopy
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DOI：
10.1117/1.2221714
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发表时间：
2006-07-01
影响因子：
3.5
通讯作者：
Lasser, Theo
中科院分区：
文献类型：
--
作者：
Leutenegger, Marcel;Blom, Hans;Lasser, Theo
我们提出了一种新的双色全内反射（TIR）荧光系统的单分子符合分析和荧光互相关光谱（FCCS）的发展和首次应用。作为性能分析，我们测量了合成的DNA结合试验，证明这种双色TIR-FCCS方法是一种合适的方法，用于测量符合测定，如生化结合，融合，或在固/液界面的信号转导。由于落射照明配置的非常高的数值孔径，我们的设置提供了非常高的荧光收集效率，导致与常规共焦FCCS相比分子亮度增加了2至3倍。通过对光谱数据的全球分析，取得了进一步的改进。(c)2006年，由光学仪器工程师协会（Society of Photo-Optical Instrumentation Engineers）主办。
We present the development and first application of a novel dual-color total internal reflection (TIR) fluorescence system for single-molecule coincidence analysis and fluorescence cross-correlation spectroscopy (FCCS). As a performance analysis, we measured a synthetic DNA-binding assay, demonstrating this dual-color TIR-FCCS approach to be a suitable method for measuring coincidence assays such as biochemical binding, fusion, or signal transduction at solid/liquid interfaces. Due to the very high numerical aperture of the epi-illumination configuration, our setup provides a very high fluorescence collection efficiency resulting in a two- to three- fold increase in molecular brightness compared to conventional confocal FCCS. Further improvements have been achieved through global analysis of the spectroscopic data. (c) 2006 Society of Photo-Optical Instrumentation Engineers.