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Proceedings Paper

Improvement of the spatial resolution in multiphoton microscopy by saturated excitation of fluorescence
Author(s): Anh Dung Nguyen; François Duport; Frédérique Vanholsbeeck; Philippe Emplit; Serge Goldman; Simon-Pierre Gorza
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Paper Abstract

We demonstrate, for the first time to the best of our knowledge, a significant resolution improvement in multiphoton microscopy (MPM) using saturated excitation of fluorescence.

At high illumination power, saturation of the two-photon excitation probability of the fluorophore occurs and induces a strong nonlinear response. By temporally modulating the excitation laser-intensity and demodulating high-order harmonics from the saturated fluorescence signal, images with higher spatial resolution than with standard MPM can be obtained. We show, as expected from a two-photon excitation process, that resolution improvement arises when demodulating at least at the third harmonic and that linear combinations of harmonics provide further improvement of the resolution. Images of 200 nm fluorescent microspheres confirm the improvement of the spatial resolution.

Paper Details

Date Published: 5 March 2015
PDF: 7 pages
Proc. SPIE 9329, Multiphoton Microscopy in the Biomedical Sciences XV, 93292G (5 March 2015); doi: 10.1117/12.2076756
Show Author Affiliations
Anh Dung Nguyen, Univ. Libre de Bruxelles (Belgium)
François Duport, Univ. Libre de Bruxelles (Belgium)
Frédérique Vanholsbeeck, The Univ. of Auckland (New Zealand)
Philippe Emplit, Univ. Libre de Bruxelles (Belgium)
Serge Goldman, Univ. Libre de Bruxelles (Belgium)
Simon-Pierre Gorza, Univ. Libre de Bruxelles (Belgium)

Published in SPIE Proceedings Vol. 9329:
Multiphoton Microscopy in the Biomedical Sciences XV
Ammasi Periasamy; Peter T. C. So; Karsten König, Editor(s)

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