Practical implementation of adaptive optics in multiphoton microscopy

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Marsh, P.N. and Burns, D. and Girkin, J.M. (2003) Practical implementation of adaptive optics in multiphoton microscopy. Optics Express, 11 (10). pp. 1123-1130. ISSN 1094-4087 (http://www.opticsinfobase.org/abstract.cfm?URI=oe-...)

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Abstract

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 µm with a resolution defined by a FWHM axial point-spread function of 1.25 µm.

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