Open Access
1 January 1996 Three-photon excitation in fluorescence microscopy
Stefan W. Hell, Karsten Bahlmann, Martin Schrader, Aleksi Soini, Henryk M. Malak, Ignacy Gryczynski, Joseph R. Lakowicz
Author Affiliations +
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.
Stefan W. Hell, Karsten Bahlmann, Martin Schrader, Aleksi Soini, Henryk M. Malak, Ignacy Gryczynski, and Joseph R. Lakowicz "Three-photon excitation in fluorescence microscopy," Journal of Biomedical Optics 1(1), (1 January 1996). https://doi.org/10.1117/12.229062
Published: 1 January 1996
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Cited by 190 scholarly publications and 2 patents.
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KEYWORDS
Luminescence

Microscopes

Microscopy

Absorption

Rhodamine

Photons

Sapphire lasers

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