8 July 2009 Fluorescence lifetime correlation spectroscopy for precise concentration detection in vivo by background subtraction
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Proceedings Volume 7368, Clinical and Biomedical Spectroscopy; 73681V (2009); doi: 10.1117/12.831572
Event: European Conferences on Biomedical Optics, 2009, Munich, Germany
Abstract
In vivo studies of single molecule dynamics by means of Fluorescence correlation spectroscopy can suffer from high background. Fluorescence lifetime correlation spectroscopy provides a tool to distinguish between signal and unwanted contributions via lifetime separation. By studying the motion of the RNA-induced silencing complex (RISC) within two compartments of a human cell, the nucleus and the cytoplasm, we observed clear differences in concentration as well as mobility of the protein complex between those two locations. Especially in the nucleus, where the fluorescence signal is very weak, a correction for background is crucial to provide reliable results of the particle number. Utilizing the fluorescent lifetime of the different contributions, we show that it is possible to distinguish between the fluorescent signal and the autofluorescent background in vivo in a single measurement.
© (2009) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.
Maria Gärtner, Jörg Mütze, Thomas Ohrt, Petra Schwille, "Fluorescence lifetime correlation spectroscopy for precise concentration detection in vivo by background subtraction", Proc. SPIE 7368, Clinical and Biomedical Spectroscopy, 73681V (8 July 2009); doi: 10.1117/12.831572; https://doi.org/10.1117/12.831572
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