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Proceedings Paper

High-speed FLIM data acquisition by time-correlated single-photon counting
Author(s): Wolfgang Becker; Axel Bergmann; Giovanni Biscotti; Karsten Koenig; Iris Riemann; Laimonas Kelbauskas; Christoph Biskup
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Paper Abstract

In this study, we describe a time-correlated single photon counting (TCSPC) technique for multi-wavelength lifetime imaging in laser-scanning microscopes. The technique is based on a four-dimensional histogramming process that records the photon density versus the time in the fluorescence decay, the x-y coordinates of the scanning area and the wavelength. It avoids any time gating or wavelength scanning and, therefore, yields a near-ideal counting efficiency. The decay functions are recorded in a large number of time channels, and the components of a multi-exponential decay can be resolved down to less than 30 ps. A single TCSPC imaging channel works with a high detection efficiency up to a photon count rate of about 5•106s-1. A modified version of the TCSPC fluorescence lifetime imaging (FLIM) technique uses several fully parallel detector and TCSPC channels. It operates at a count rate of more than 107 photons per second and records double-exponential FLIM data within less than 10 seconds.

Paper Details

Date Published: 21 June 2004
PDF: 9 pages
Proc. SPIE 5323, Multiphoton Microscopy in the Biomedical Sciences IV, (21 June 2004); doi: 10.1117/12.529113
Show Author Affiliations
Wolfgang Becker, Becker and Hickl GmbH (Germany)
Axel Bergmann, Becker and Hickl GmbH (Germany)
Giovanni Biscotti, Becker and Hickl GmbH (Germany)
Karsten Koenig, Fraunhofer Institut fur Biomedizinische Technik (Germany)
Iris Riemann, Fraunhofer Institut fur Biomedizinische Technik (Germany)
Laimonas Kelbauskas, Friedrich Schiller Univ. Jena (Germany)
Christoph Biskup, Friedrich Schiller Univ. Jena (Germany)

Published in SPIE Proceedings Vol. 5323:
Multiphoton Microscopy in the Biomedical Sciences IV
Ammasi Periasamy; Peter T. C. So, Editor(s)

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