Author(s): F. Cella Zanacchi; Z. Lavagnino; M. Pesce; F. Difato; E. Ronzitti; A. Diaspro.
Light-sheet microscopy, such as ultramicroscopy, single plane illumination microscopy (SPIM)  and digital scanned laser microscopy (DSLM) , represents a useful tool for biological investigations of thick samples. Such techniques have been found particularly useful in developmental biology applications since they provide the capability to perform fast imaging of living samples reducing photobleaching effects. The high signal to noise ratio and the intrinsic optical sectioning capability provided by SPIM suggest this technique as the best choice for imaging of thick scattering samples. Nevertheless, imaging in depth of large samples suffers from a decreasing in the image quality due to scattering effects. Two photon excitation microscopy  became a popular tool to perform imaging in turbid media since it improves the penetration depth capability and it reduces the image quality degradation due to scattering  and light matter interactions. Therefore, two photon excitation within the light sheet illumination scheme has been exploited in order to reduce scattering effects due to light-sample interactions. In this work two photon excitation imaging in SPIM scheme has been performed in order to achieve an improvement in the penetration depth while imaging living biological samples.