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Two-photon excitation selective plane illumination microscopy (2PE-SPIM) of highly scattering samples: characterization and application

Two-photon excitation selective plane illumination microscopy (2PE-SPIM) of highly scattering samples: characterization and application

Optics Express 21(5): 5998-6008

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.

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Accession: 056723615

Download citation: RISBibTeXText

PMID: 23482168

DOI: 10.1364/oe.21.005998

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