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.

© 2011 Optical Society of America

Corkscrew point spread function for far-field three-dimensional nanoscale localization of pointlike objects

Matthew D. Lew, Steven F. Lee, Majid Badieirostami, and W. E. Moerner
Opt. Lett. 36(2) 202-204 (2011)

Simple way of pinpointing the three-dimensional position of biomarkers in fluorescence microscopy using a through-focus exposure method

Koichiro Kishima
Appl. Opt. 50(25) 4989-4997 (2011)

Two-photon fluorescence isotropic-single-objective microscopy

Eric Le Moal, Emeric Mudry, Patrick C. Chaumet, Patrick Ferrand, and Anne Sentenac
Opt. Lett. 37(1) 85-87 (2012)

Three-photon fluorescence microscopy with an axially elongated Bessel focus

Cristina Rodríguez, Yajie Liang, Rongwen Lu, and Na Ji
Opt. Lett. 43(8) 1914-1917 (2018)

Cellular resolution optical coherence microscopy with high acquisition speed for in-vivo human skin volumetric imaging

Kye-Sung Lee, Kevin P. Thompson, Panomsak Meemon, and Jannick P. Rolland
Opt. Lett. 36(12) 2221-2223 (2011)