Widefield TSCSPC-systems with large-area-detectors: application in simultaneous multi-channel-FLIM

Sergei Stepanov; Sergei Bakhlanov; Evgeny Drobchenko; Hann-Jörg Eckert; Klaus Kemnitz

doi:10.1117/12.871523

24 November 2010 Widefield TSCSPC-systems with large-area-detectors: application in simultaneous multi-channel-FLIM

Sergei Stepanov, Sergei Bakhlanov, Evgeny Drobchenko, Hann-Jörg Eckert, Klaus Kemnitz

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.

Citation Download Citation

Sergei Stepanov, Sergei Bakhlanov, Evgeny Drobchenko, Hann-Jörg Eckert, and 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); https://doi.org/10.1117/12.871523

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