Development of a confocal laser scanning fluorescence microscope using two-photon excitation in combination with time-gated detection

Joost Sytsma; Jurrien Vroom; Hans C. Gerritsen; Yehudi K. Levine

doi:10.1117/12.205328

23 March 1995 Development of a confocal laser scanning fluorescence microscope using two-photon excitation in combination with time-gated detection

Joost Sytsma, Jurrien Vroom, Hans C. Gerritsen, Yehudi K. Levine

Author Affiliations +

Proceedings Volume 2412, Three-Dimensional Microscopy: Image Acquisition and Processing II; (1995) https://doi.org/10.1117/12.205328
Event: IS&T/SPIE's Symposium on Electronic Imaging: Science and Technology, 1995, San Jose, CA, United States

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.

Citation Download Citation

Joost Sytsma, Jurrien Vroom, Hans C. Gerritsen, and Yehudi K. Levine "Development of a confocal laser scanning fluorescence microscope using two-photon excitation in combination with time-gated detection", Proc. SPIE 2412, Three-Dimensional Microscopy: Image Acquisition and Processing II, (23 March 1995); https://doi.org/10.1117/12.205328

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