Fluorescence lifetime imaging of cells on the picosecond timescale

Klaus Kemnitz; Rene Paul; Jacques Coppey; Maite Coppey-Moisan

doi:10.1117/12.260795

6 December 1996 Fluorescence lifetime imaging of cells on the picosecond timescale

Klaus Kemnitz, Rene Paul, Jacques Coppey, Maite Coppey-Moisan

Proceedings Volume 2926, Optical Biopsies and Microscopic Techniques; (1996) https://doi.org/10.1117/12.260795
Event: BiOS Europe '96, 1996, Vienna, Austria

Abstract

Novel MCP detectors for time- and space-correlated single photon counting (TSCSPC) spectroscopy, featuring delay-line (DL) or quadrant anode (QA), are employed in microscopic fluorescence lifetime imaging on the picosecond time scale. The linear DL-MCP-PMT is characterized by a spatial instrument response function (IRF) of 100 micrometer FWHM, resulting in 200 space channels, whereas the QA-MCP-PMT is a 2D imager with 400 by 400 pixel at 40 micrometer resolution. The detectors have a temporal IRF of 75 ps (DL) and 120 ps (QA) FWHM, sufficient for 10 ps time resolution. First results on TOTO-fixed cell systems are presented, demonstrating high-quality kinetics at subcellular resolution, with up to 6 lifetime species at higher dye concentrations, characteristically distributed among individual cell compartments. A comparison with TOTO/DNA- suspensions is made that serve as reference system.

Citation Download Citation

Klaus Kemnitz, Rene Paul, Jacques Coppey, and Maite Coppey-Moisan "Fluorescence lifetime imaging of cells on the picosecond timescale", Proc. SPIE 2926, Optical Biopsies and Microscopic Techniques, (6 December 1996); https://doi.org/10.1117/12.260795

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