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Journal of Biomedical Optics

Three-photon excitation in fluorescence microscopy
Author(s): Stefan W. Hell; Karsten Bahlmann; Martin Schrader; Aleksi Soini; Henryk M. Malak; Ignacy Gryczynski; Joseph R. Lakowicz
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Paper Abstract

We show experiments proving the feasibility of scanning fluorescence microscopy by three-photon excitation. Three-photon excitation fluorescence axial images are shown of polystyrene beads stained with the fluorophore 2,5-bis(4-biphenyl)oxazole (BBO). Three-photon excitation is performed at an excitation wavelength of 900 nm and with pulses of 130 fs duration provided by a mode-locked titanium sapphire laser. Fluorescence is collected between 350 and 450 nm. The fluorescence image signal features a third-order dependence on the excitation power, also providing intrinsic 3-D imaging. The resolution of a three-photon excitation microscope is increased over that of a comparable two-photon excitation microscope.

Paper Details

Date Published: 2 January 1996
PDF: 4 pages
J. Biomed. Opt. 1(1) doi: 10.1117/12.229062
Published in: Journal of Biomedical Optics Volume 1, Issue 1
Show Author Affiliations
Stefan W. Hell, Univ. of Turku (Finland)
Karsten Bahlmann, Univ. of Turku (Finland)
Martin Schrader, Univ. of Turku (Finland)
Aleksi Soini, Univ. of Turku (Finland)
Henryk M. Malak, Univ. of Maryland/Baltimore School of Medicine (United States)
Ignacy Gryczynski, Univ. of Maryland School of Medicine (United States)
Joseph R. Lakowicz, Univ. of Maryland/Baltimore School of Medicine (United States)

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