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Proceedings Paper

Lifetime imaging with the Zeiss LSM-510
Author(s): Wolfgang Becker; Axel Bergmann; Georg Weiss
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Paper Abstract

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.

Paper Details

Date Published: 17 June 2002
PDF: 6 pages
Proc. SPIE 4620, Multiphoton Microscopy in the Biomedical Sciences II, (17 June 2002); doi: 10.1117/12.470706
Show Author Affiliations
Wolfgang Becker, Becker and Hickl GmbH (Germany)
Axel Bergmann, Becker and Hickl GmbH (Germany)
Georg Weiss, Carl Zeiss Jena GmbH (Germany)

Published in SPIE Proceedings Vol. 4620:
Multiphoton Microscopy in the Biomedical Sciences II
Ammasi Periasamy; Peter T. C. So, Editor(s)

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