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Adaptive optics in confocal and two-photon microscopy of rat brain: a single correction per optical section

Girkin, J.M. and Vijverberg, J. and Orazio, M.A. and Poland, S.P. and Wright, A. and Cobb, S. (2007) Adaptive optics in confocal and two-photon microscopy of rat brain: a single correction per optical section. Proceedings of SPIE: The International Society for Optical Engineering, 6442. 64420T.

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Abstract

Sample induced optical aberrations in slices of rat brain tissue have been corrected with a deformable membrane mirror. The aberration correction required by the DMM was determined using a genetic algorithm with the intensity at a point in the sample as a fitness value. We show that by optimising on the intensity of a single point in the sample we are able to improve the axial resolution across the whole field of view of the image at a fixed sample depth. The ratio between the corrected axial resolution and the diffraction limited resolution is on average 2.7 for a 50 µm thick rat brain tissue sample and 12 for a 380 µm thick sample across the whole field of view. The uncorrected ratio being 4.1 and 15.5 respectively. Using a single aberration correction per depth, compared to a point-by-point aberration correction, will significantly decrease scan times and therefore reduce photobleaching and phototoxic effects enabling more rapid microscopy with active aberration correction.