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Practical implementation of adaptive optics in multiphoton microscopy

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

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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.