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
Abstract 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.
Silvestri Ludovico、Gheisari Ali、Sacconi Leonardo、Pavone Francesco S.、Vanzi Francesco、Costantini Irene、Turrini Lapo、M¨1llenbroich Caroline M.、Alterini Tommaso、Di Giovanna Antonino P.
National Institute of Optics, National Research Council||European Laboratory for Non-Linear SpectroscopyEuropean Laboratory for Non-Linear SpectroscopyNational Institute of Optics, National Research Council||European Laboratory for Non-Linear SpectroscopyNational Institute of Optics, National Research Council||European Laboratory for Non-Linear Spectroscopy||Department of Physics and Astronomy, University of FlorenceEuropean Laboratory for Non-Linear Spectroscopy||Department of Biology, University of FlorenceEuropean Laboratory for Non-Linear Spectroscopy||University of FlorenceEuropean Laboratory for Non-Linear SpectroscopyNational Institute of Optics, National Research Council||European Laboratory for Non-Linear SpectroscopyEuropean Laboratory for Non-Linear SpectroscopyEuropean Laboratory for Non-Linear Spectroscopy||University of Florence
生物物理学光电子技术
Silvestri Ludovico,Gheisari Ali,Sacconi Leonardo,Pavone Francesco S.,Vanzi Francesco,Costantini Irene,Turrini Lapo,M¨1llenbroich Caroline M.,Alterini Tommaso,Di Giovanna Antonino P..Increasing sensitivity and accuracy of brain-wide quantitative studies in light-sheet microscopy[EB/OL].(2025-03-28)[2025-04-30].https://www.biorxiv.org/content/10.1101/230540.点此复制
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