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.
© 2011 Optical Society of America
Fourier Optics and Signal Processing
Original Manuscript: July 22, 2011
Manuscript Accepted: August 8, 2011
Published: September 1, 2011
Vol. 6, Iss. 10 Virtual Journal for Biomedical Optics
Marco Dal Maschio, Angela Michela De Stasi, Fabio Benfenati, and Tommaso Fellin, "Three-dimensional in vivo scanning microscopy with inertia-free focus control," Opt. Lett. 36, 3503-3505 (2011)