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Proceedings Paper

Fluorescence lifetime correlation spectroscopy for precise concentration detection in vivo by background subtraction
Author(s): Maria Gärtner; Jörg Mütze; Thomas Ohrt; Petra Schwille
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Paper 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.

Paper Details

Date Published: 7 July 2009
PDF: 7 pages
Proc. SPIE 7368, Clinical and Biomedical Spectroscopy, 73681V (7 July 2009); doi: 10.1117/12.831572
Show Author Affiliations
Maria Gärtner, BIOTEC, Dresden Univ. of Technology (Germany)
Jörg Mütze, BIOTEC, Dresden Univ. of Technology (Germany)
Thomas Ohrt, BIOTEC, Dresden Univ. of Technology (Germany)
Petra Schwille, BIOTEC, Dresden Univ. of Technology (Germany)

Published in SPIE Proceedings Vol. 7368:
Clinical and Biomedical Spectroscopy
Irene Georgakoudi; Jürgen Popp; Katarina Svanberg, Editor(s)

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