title: Two-photon excitation selective plane illumination microscopy (2PE-SPIM) of highly scattering samples: characterization and application.
abstract: In this work we report the advantages provided by two photon excitation (2PE) implemented in a selective plane illumination microscopy (SPIM) when imaging thick scattering samples. In particular, a detailed analysis of the effects induced on the real light sheet excitation intensity distribution is performed. The comparison between single-photon and two-photon excitation profiles shows the reduction of the scattering effects and sample-induced aberrations provided by 2PE-SPIM. Furthermore, uniformity of the excitation distribution and the consequent improved image contrast is shown when imaging scattering phantom samples in depth by 2PE-SPIM. These results show the advantages of 2PE-SPIM and suggest how this combination can further enhance the SPIM performance. Phantom samples have been designed with optical properties compatible with biological applications of interest.
authors.affiliation: Department of Physics, University of Genoa, Via Dodecaneso 33, 16146 Genoa, Italy.
authors.forename: Zeno
authors.initials: Z
authors.lastname: Lavagnino
authors.forename: Francesca Cella
authors.initials: FC
authors.lastname: Zanacchi
authors.forename: Emiliano
authors.initials: E
authors.lastname: Ronzitti
authors.forename: Alberto
authors.initials: A
authors.lastname: Diaspro
mesh_terms: D004847:Epithelial Cells / Q000166:cytology; D006801:Humans; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods*; D019047:Phantoms, Imaging; D012542:Scattering, Radiation