1 January 1996 Three-photon excitation in fluorescence microscopy
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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, Stefan W. Hell, Karsten Bahlmann, Karsten Bahlmann, Martin Schrader, Martin Schrader, Aleksi Soini, Aleksi Soini, Henryk M. Malak, Henryk M. Malak, Ignacy Gryczynski, Ignacy Gryczynski, Joseph R. Lakowicz, 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 . Submission:

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