Three-dimensional in vivo scanning microscopy with inertia-free focus control.

Opt Lett 2011 Sep;36(17):3503-5

Department of Neuroscience and Brain Technologies, Istituto Italiano di Tecnologia, Genova, Italy.

The acquisition of high-resolution images in three dimensions is of utmost importance for the morphological and functional investigation of biological tissues. Here, we present a laser scanning two-photon microscope with remote and motionless control of the focus position. The movement of the excitation spot along the propagation direction is achieved by shaping the laser wavefront with a spatial light modulator. Depending on the optical properties of the objective in use, this approach allows z movements in a range of tens to hundreds of micrometers with small changes of the point spread function. We applied this technique for the three-dimensional (3D) imaging of fluorescent cells in the mouse neocortex in vivo. The presented system bypasses the limitations of microscopes based on moving objectives, enabling high-resolution inertia-free 3D imaging.

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http://dx.doi.org/10.1364/OL.36.003503DOI Listing
September 2011
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