Simultaneous two-photon imaging and photostimulation with structured light illumination.

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  • 04 April 2012
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Holographic microscopy is increasingly recognized as a promising tool for the study of the central nervous system. Here we present a “holographic module”, a simple optical path that can be combined  with commercial scanheads for simultaneous imaging and uncaging with structured two-photon light. The present microscope is coupled to two independently tunable lasers and has two principal configurations: holographic imaging combined with galvo-steered uncaging and holographic uncaging combined with conventional scanning imaging. We applied this flexible system for simultaneous two-photon imaging and photostimulation of neuronal cells with complex light patterns, opening new perspectives for the study of brain function in situ and in vivo.