A demonstration of the effectiveness of a single aberration correction per optical slice in beam scanned optically sectioning microscopes

In this paper we report the use of adaptive optics to correct for sample induced aberrations in optical microscopy, crucially comparing individual pixel-by-pixel correction against a single correction for an entire optical section. Sample induced optical aberrations in slices of rat brain tissue were corrected with a deformable membrane mirror. Using axial resolution measurements, we demonstrate that a single aberration correction per optical slice achieves around 80% of the maximum possible improvement compared to individual pixel-by-pixel correction in both confocal and multiphoton microscopy. A single aberration correction per depth, compared to pixel-by-pixel aberration correction, significantly decreases scan times and therefore reduces photobleaching and phototoxic effects enabling more rapid microscopy with active aberration correction. The results confirm that the use of a “look-up” table, based upon sample type and depth, may be the most practical way of implementing adaptive optic aberration correction in beam scanning optical sectioning microscopy.