Open AccessPractical implementation of adaptive optics in multiphoton microscopy
Author(s) -
Paul N. Marsh,
D. Burns,
John M. Girkin
Publication year - 2003
Publication title -
optics express
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.394
H-Index - 271
ISSN - 1094-4087
DOI - 10.1364/oe.11.001123
Subject(s) - optics , adaptive optics , wavefront , deformable mirror , point spread function , refractive index , microscopy , materials science , microscope , laser , physics
A dedicated two-photon microscope incorporating adaptive-optic correction of specimen-induced aberrations is presented. Wavefront alteration of the scanning laser beam was achieved by use of a micromachined deformable mirror. Post scan head implementation produces a compact module compatible with the Bio-Rad MRC-600 scan head. Automatic aberration correction using feedback from the multiphoton fluorescence intensity allowed the adaptive optic to extend the imaging depth attainable in both artificial and biological refractive-index mismatched samples. With a 1.3-NA, x40, Nikon oil immersion objective, the imaging depth in water was extended from approximately 3.4 to 46.2 microm with a resolution defined by a FWHM axial point-spread function of 1.25 microm.