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Proceedings Paper

FRET measurements by TCSPC laser scanning microscopy
Author(s): Wolfgang Becker; Klaus Benndorf; Axel Bergmann; Christoph Biskup; Karsten Koenig; Uday Tirlapur; Thomas Zimmer
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Paper Abstract

We use a two-photon laser scanning microscope with a new Time-Correlated Single Photon Counting (TCSPC) imaging technique to obtain combined intensity-lifetime images for FRET measurements in living cells. Single photon pulses from a photomultiplier and signals from the scanning head are used to record the three-dimensional photon density over the time- and image coordinates. Double exponential decay analysis delivers the lifetime components of the quenched and the unquenched molecules in all pixels of the image. We use the ratio of the intensity coefficients of the fast and slow decay component to create images that show the size of the FRET effects in different parts of the cell.

Paper Details

Date Published: 2 November 2001
PDF: 5 pages
Proc. SPIE 4431, Photon Migration, Optical Coherence Tomography, and Microscopy, (2 November 2001); doi: 10.1117/12.447406
Show Author Affiliations
Wolfgang Becker, Becker and Hickl GmbH (Germany)
Klaus Benndorf, Friedrich-Schiller-Univ. Jena (Germany)
Axel Bergmann, Becker and Hickl GmbH (Germany)
Christoph Biskup, Friedrich-Schiller-Univ. Jena (Germany)
Karsten Koenig, Friedrich-Schiller-Univ. Jena (Germany)
Uday Tirlapur, Friedrich-Schiller-Univ. Jena (Germany)
Thomas Zimmer, Friedrich-Schiller-Univ. Jena (France)

Published in SPIE Proceedings Vol. 4431:
Photon Migration, Optical Coherence Tomography, and Microscopy
Stefan Andersson-Engels; Michael F. Kaschke, Editor(s)

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