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.
We measure the form of the point spread function in the three main configurations of 4 Pi microscopy as well as the traditional confocal arrangement. We confirm, for the first time experimentally, the equivalence between the 4 Pi (A) and 4 Pi (B) geometries. We measure a 6.9 fold increase in axial resolution over the confocal case.
Conference Committee Involvement (9)
Optics, Photonics and Digital Technologies for Imaging Applications VII
6 April 2022 | Strasbourg, France
Optics, Photonics and Digital Technologies for Imaging Applications VI
6 April 2020 | Online Only, France
Optics, Photonics and Digital Technologies for Imaging Applications
24 April 2018 | Strasbourg, France
Optics, Photonics and Digital Technologies for Imaging Applications
5 April 2016 | Brussels, Belgium
Optics, Photonics and Digital Technologies for Multimedia Applications
16 April 2014 | Brussels, Belgium
Optics, Photonics and Digital Technologies for Multimedia Applications
17 April 2012 | Brussels, Belgium
Optics, Photonics and Digital Technologies for Multimedia Applications
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