24 November 2010 Widefield TSCSPC-systems with large-area-detectors: application in simultaneous multi-channel-FLIM
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Proceedings Volume 7376, Laser Applications in Life Sciences; 73760Z (2010) https://doi.org/10.1117/12.871523
Event: Laser Applications in Life Sciences 2010, 2010, Oulu, Finland
Abstract
Novel proximity-type Time- and Space-Correlated Single Photon Counting (TSCSPC) crossed-delay-line (DL)- and multi-anode (MA)-systems of outstanding performance and homogeneity were developed, using large-area detector heads of 25 and 40 mm diameter. Instrument response functions IRF(space) = (60 ± 5) μm FWHM and IRF(time) = (28 ± 3) ps FWHM were achieved over the full 12 cm2 area of the detector. Deadtime at throughput of 105 cps is 10% for "high-resolution" system and 5% in the "video"-system at 106 cps, at slightly reduced time- and space resolution. A fluorescence lifetime of (3.5 ± 1) ps can be recovered from multi-exponential dynamics of a single living cyanobacterium (Acaryochloris marina). The present large-area detectors are particularly useful in simultaneous multichannel applications, such as 2-colour anisotropy or 4-colour lifetime imaging, utilizing dual- or quad-view image splitters. The long-term stability, low- excitation-intensity (< 100 mW/cm2) widefield systems enable minimal-invasive observation, without significant bleaching or photodynamic reactions, thus allowing long-period observation of up to several hours in living cells.
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Sergei Stepanov, Sergei Stepanov, Sergei Bakhlanov, Sergei Bakhlanov, Evgeny Drobchenko, Evgeny Drobchenko, Hann-Jörg Eckert, Hann-Jörg Eckert, Klaus Kemnitz, Klaus Kemnitz, } "Widefield TSCSPC-systems with large-area-detectors: application in simultaneous multi-channel-FLIM", Proc. SPIE 7376, Laser Applications in Life Sciences, 73760Z (24 November 2010); doi: 10.1117/12.871523; https://doi.org/10.1117/12.871523
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