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Proceedings Paper

New method for fluorescence lifetime imaging in bilateral confocal microscopy by double-pulse excitation
Author(s): G. J. Brakenhoff; Michiel Mueller; Rick I. Ghauharali; Koen Visscher
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Paper Abstract

A new technique for the measurement of fluorescence lifetimes relies on the (near steady state) excitation with short optical pulses. The novel technique has the potentiality to provide high time resolution--since it relies on the laser pulse duration, rather than on electronic gating techniques--and permits, in combination with bilateral confocal microscopy and the use of a (cooled) CCD, sensitive signal detection over a large dynamic range. Combined with confocal microscopy it enables the spatial determination of the fluorescence lifetimes, the value of which is influenced by the local environment of fluorescent probe molecules in biological samples. The principle of the technique is discussed within a theoretical framework taking into account various secondary effects.

Paper Details

Date Published: 23 March 1995
PDF: 9 pages
Proc. SPIE 2412, Three-Dimensional Microscopy: Image Acquisition and Processing II, (23 March 1995); doi: 10.1117/12.205329
Show Author Affiliations
G. J. Brakenhoff, Univ. of Amsterdam (Netherlands)
Michiel Mueller, Univ. of Amsterdam (Netherlands)
Rick I. Ghauharali, Univ. of Amsterdam (Netherlands)
Koen Visscher, Princeton Univ. (United States)


Published in SPIE Proceedings Vol. 2412:
Three-Dimensional Microscopy: Image Acquisition and Processing II
Tony Wilson; Carol J. Cogswell, Editor(s)

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