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

John M. Girkin; J. Vijverberg; M. Orazio; S. Poland; A. J. Wright

doi:10.1117/12.696761

10 February 2007 Adaptive optics in confocal and two-photon microscopy of rat brain: a single correction per optical section

John M. Girkin, J. Vijverberg, M. Orazio, S. Poland, A. J. Wright

Author Affiliations +

Proceedings Volume 6442, Multiphoton Microscopy in the Biomedical Sciences VII; 64420T (2007) https://doi.org/10.1117/12.696761
Event: SPIE BiOS, 2007, San Jose, California, United States

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.

Citation Download Citation

John M. Girkin, J. Vijverberg, M. Orazio, S. Poland, and A. J. Wright "Adaptive optics in confocal and two-photon microscopy of rat brain: a single correction per optical section", Proc. SPIE 6442, Multiphoton Microscopy in the Biomedical Sciences VII, 64420T (10 February 2007); https://doi.org/10.1117/12.696761

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