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Proceedings Paper

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

In this study, we present two different approaches that allow multi-wavelength fluorescence lifetime measurements in the time domain. One technique is based on a streak camera system, the other technique is based on a time-correlated singlephoton- counting (TCSPC) approach. The setup consists of a confocal laser-scanning microscope (LSM 510, Zeiss) and a Titanium:Sapphire-laser (Mira 900D, Coherent) that is used for pulsed one- and two-photon excitation. Fluorescence light emitted by the sample is dispersed by a polychromator (250is, Chromex) and recorded by a streak camera (C5680 with M5677 sweep unit, Hamamatsu Photonics) or a 16 channel TCSPC detector head (PML-16, Becker & Hickl) connected to a TCSPC imaging module (SPC-730/SPC-830, Becker & Hickl). 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 over fluorescence techniques that are already well established in the field of confocal microscopy, such as spectrally resolved intensity measurements or single-wavelength fluorescence lifetime measurements.

Paper Details

Date Published: 10 February 2007
PDF: 12 pages
Proc. SPIE 6442, Multiphoton Microscopy in the Biomedical Sciences VII, 64420V (10 February 2007); doi: 10.1117/12.699870
Show Author Affiliations
Christoph Biskup, Friedrich-Schiller-Univ. Jena (Germany)
Birgit Hoffmann, Friedrich-Schiller-Univ. Jena (Germany)
Laimonas Kelbauskas, Friedrich-Schiller-Univ. Jena (Germany)
Biodesign Institute, Arizona State Univ. (United States)
Thomas Zimmer, Friedrich-Schiller-Univ. Jena (Germany)
Sascha Dietrich, Friedrich-Schiller-Univ. Jena (Germany)
Wolfgang Becker, Becker & Hickl GmbH (Germany)
Axel Bergmann, Becker & Hickl GmbH (Germany)
Nikolaj Klöcker, Albert-Ludwigs-Univ. Freiburg (Germany)
Klaus Benndorf, Friedrich-Schiller-Univ. Jena (Germany)


Published in SPIE Proceedings Vol. 6442:
Multiphoton Microscopy in the Biomedical Sciences VII
Ammasi Periasamy; Peter T. C. So, Editor(s)

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