Open AccessTwo-photon excitation selective plane illumination microscopy (2PE-SPIM) of highly scattering samples: characterization and application
Author(s) -
Zeno Lavagnino,
Francesca Cella Zanacchi,
Emiliano Ronzitti,
Alberto Diaspro
Publication year - 2013
Publication title -
optics express
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.394
H-Index - 271
ISSN - 1094-4087
DOI - 10.1364/oe.21.005998
Subject(s) - optics , imaging phantom , excitation , scattering , materials science , microscopy , two photon excitation microscopy , light scattering , photon , optoelectronics , physics , fluorescence , quantum mechanics
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.