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17 June 2002 Lifetime imaging with the Zeiss LSM-510
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Proceedings Volume 4620, Multiphoton Microscopy in the Biomedical Sciences II; (2002)
Event: International Symposium on Biomedical Optics, 2002, San Jose, CA, United States
The Zeiss LSM-510 NLO laser scanning microscope can be combined with a new TCSPC (time-correlated single photon counting) lifetime imaging technique developed by Becker & Hickl, Berlin. This technique is based on a three-dimensional histogramming process that records the photon density over the time within the fluorescence decay function and the x-y coordinates of the scanning area. The histogramming process avoids any time gating and therefore yields a counting efficiency close to one. Upgrading The LSM-510 for TCSPC imaging does not require changes in the microscope hardware or software. A fast detector is attached to the fibre output of the scanning head, and synchronisation of the TCSPC module with scanning is achieved via the user I/O of the scan controller. With an MCP-PMT as a detector, fluorescence decay components down to 30 ps can be resolved. The capability of the instrument is shown for the separation of chromphores by their fluorescence lifetime and for CFP/YFP FRET.
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Wolfgang Becker, Axel Bergmann, and Georg Weiss "Lifetime imaging with the Zeiss LSM-510", Proc. SPIE 4620, Multiphoton Microscopy in the Biomedical Sciences II, (17 June 2002);

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