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Increasing sensitivity and accuracy of brain-wide quantitative studies in light-sheet microscopy
Increasing sensitivity and accuracy of brain-wide quantitative studies in light-sheet microscopy
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Increasing sensitivity and accuracy of brain-wide quantitative studies in light-sheet microscopy
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Increasing sensitivity and accuracy of brain-wide quantitative studies in light-sheet microscopy
Increasing sensitivity and accuracy of brain-wide quantitative studies in light-sheet microscopy
Paper

Increasing sensitivity and accuracy of brain-wide quantitative studies in light-sheet microscopy

2017
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Overview
Light-sheet microscopy (LSM) has proven a useful tool in neuroscience to image whole brains with high frame rates at cellular resolution. LSM is employed either in combination with tissue clearing to reconstruct the cyto-architecture over the entire mouse brain or with intrinsically transparent samples like zebrafish larvae for functional imaging. Inherently to LSM, however, residual opaque objects cause stripe artifacts, which obscure features of interest and, during functional imaging, modulate fluorescence variations related to neuronal activity. Here, we report how Bessel beams reduce streaking artifacts and produce high-fidelity structural data. Furthermore, using Bessel beams, we demonstrate a fivefold increase in sensitivity to calcium transients and a 20-fold increase in accuracy in the detection of activity correlations in functional imaging. Our results demonstrate the contamination of data by systematic and random errors through Gaussian illumination and furthermore quantify the increase in fidelity of such data when using Bessel beams.
Publisher
Cold Spring Harbor Laboratory Press,Cold Spring Harbor Laboratory