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Proceedings Paper

Multidimensional fluorescence lifetime measurements
Author(s): Christoph Biskup; Birgit Hoffmann; Laimonas Kelbauskas; Thomas Zimmer; Nikolaj Klöcker; Wolfgang Becker; Klaus Benndorf
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Paper Abstract

In this study, we present two different approaches that can be used for multi-wavelength fluorescence lifetime measurements in the time domain. One technique is based on a streak-camera system, the other technique is based on the time-correlated-single-photon-counting (TCSPC) approach. The setup consists of a confocal laser-scanning microscope and a Titanium:Sapphire-laser that is used for pulsed one- and two-photon excitation. Fluorescence light emitted by the sample is fed back through the scan head and guided to one of the confocal channels, where it is coupled into an optical fiber and directed to a polychromator. The polychromator disperses the emitted light according to its wavelength and focuses the resulting spectrum on the entrance slit of a streak camera or a 16 channel PMT array, which is connected to a TCSPC imaging module. With these techniques it is possible to acquire fluorescence decays in several wavelength regions simultaneously. We applied these methods to Förster resonance energy transfer (FRET) measurements and discuss the advantages and pitfalls of fluorescence lifetime measurements.

Paper Details

Date Published: 15 February 2008
PDF: 12 pages
Proc. SPIE 6860, Multiphoton Microscopy in the Biomedical Sciences VIII, 68601I (15 February 2008); doi: 10.1117/12.762834
Show Author Affiliations
Christoph Biskup, Friedrich-Schiller-Univ. (Germany)
Birgit Hoffmann, Friedrich-Schiller-Univ. (Germany)
Laimonas Kelbauskas, Friedrich-Schiller-Univ. (Germany)
Arizona State Univ. (United States)
Thomas Zimmer, Friedrich-Schiller-Univ. (Germany)
Nikolaj Klöcker, Albert-Ludwigs-Univ. Freiburg (Germany)
Wolfgang Becker, Becker & Hickl GmbH (Germany)
Klaus Benndorf, Friedrich-Schiller-Univ. (Germany)


Published in SPIE Proceedings Vol. 6860:
Multiphoton Microscopy in the Biomedical Sciences VIII
Ammasi Periasamy; Peter T. C. So, Editor(s)

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