Two-Photon Excitation for Optical Nanoscopy and Light-Sheet Illumination Microscopy
We report about coupling two-photon excitation (2PE) with two, comparatively young and successful optical methods specially realized for the improvement of resolution and 3D imaging of large samples, i.e. STED
(stimulated emission depletion) and SPI (selective plane illumination) microscopy. In the former case we aimed to get a better resolution for 2PE microscopy while for the latter we aimed to obtain better penetration in scattering samples by shifting the light sheet wavelengths to the red in the non-linear domain. We show results obtained by adapting a commercial STED-CW microscope to 2PE and by implementing a 2PE-SPIM homemade system. The implementation of these systems is demonstrated by the imaging of living Saccharomyces cervisiae cells.
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