A custom two-photon and second-harmonic microscope

The introduction of two-photon microscopy has revolutionized life sciences by enabling long-term imaging of living preparations in highly scattering tissue while minimizing photodamage. At the same time, commercial two-photon microscopes are expensive and this has prevented the widespread distribution of this technique to the biological community. As an alternative to commercial systems, we provide an update of our efforts designing custom-built two-photon instruments by modifying the Olympus Fluoview laser scanning confocal microscope. With the newer version of our instrument we modulate the intensity of the laser beam using a Pockel's cell in arbitrary spatiotemporal patterns, perform simultaneous optical imaging and optical stimulation experiments and also can combine them with second harmonic generation measurements.

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