title: Three-dimensional in vivo scanning microscopy with inertia-free focus control.
abstract: 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.
authors: Department of Neuroscience and Brain Technologies, Istituto Italiano di Tecnologia, Genova, Italy.
authors: Marco
authors: M
authors: Dal Maschio
authors: Angela Michela
authors: AM
authors: De Stasi
authors: Fabio
authors: F
authors: Benfenati
authors: Tommaso
authors: T
authors: Fellin
mesh_terms: D000818:Animals; D001921:Brain / Q000166:cytology; D021621:Imaging, Three-Dimensional / Q000379:methods*; D008027:Light; D051379:Mice; D008853:Microscopy / Q000379:methods*; D055620:Optical Phenomena; D017785:Photons