21 June 2004 High-speed FLIM data acquisition by time-correlated single-photon counting
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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.
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Wolfgang Becker, Wolfgang Becker, Axel Bergmann, Axel Bergmann, Giovanni Biscotti, Giovanni Biscotti, Karsten Koenig, Karsten Koenig, Iris Riemann, Iris Riemann, Laimonas Kelbauskas, Laimonas Kelbauskas, Christoph Biskup, Christoph Biskup, } "High-speed FLIM data acquisition by time-correlated single-photon counting", Proc. SPIE 5323, Multiphoton Microscopy in the Biomedical Sciences IV, (21 June 2004); doi: 10.1117/12.529113; https://doi.org/10.1117/12.529113
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