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Proceedings Paper

Development of a confocal laser scanning fluorescence microscope using two-photon excitation in combination with time-gated detection
Author(s): Joost Sytsma; Jurrien Vroom; Hans C. Gerritsen; Yehudi K. Levine
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Paper Abstract

Fluorescent molecules having single-photon absorption in the blue and the UV can be excited with infra-red light via a process known as two-photon excitation. The combination of this technique with scanning techniques can be exploited for 3D microscopic imaging. The two- photon process is confined to a restricted volume in the sample determined by the laser focus, resulting in inherent confocality. Other advantages are reduced photo-bleaching of the samples and a larger penetration depth of the excitation light. The implementation of time-gated detection techniques allows fluorescent lifetime imaging. This drastically improves the selectivity and contrast of the images.

Paper Details

Date Published: 23 March 1995
PDF: 5 pages
Proc. SPIE 2412, Three-Dimensional Microscopy: Image Acquisition and Processing II, (23 March 1995); doi: 10.1117/12.205328
Show Author Affiliations
Joost Sytsma, Univ. Utrecht (Netherlands)
Jurrien Vroom, Univ. Utrecht (Netherlands)
Hans C. Gerritsen, Univ. Utrecht (Netherlands)
Yehudi K. Levine, Univ. Utrecht (Netherlands)

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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