Light-sheet microscopy (LSM) has proven a useful tool in neuroscience to image whole brains with high frame rates at cellular resolution and, in combination with tissue clearing methods, is often employed to reconstruct the cyto-architecture over the intact mouse brain. Inherently to LSM, however, residual opaque objects, always present to some extent even in extremely well optically cleared samples, cause stripe artifacts, which, in the best case, severely affect image homogeneity and, in the worst case, completely obscure features of interest. Here, demonstrating two example applications in intact optically cleared mouse brains, we report how Bessel beams reduce streaking artifacts and produce high-fidelity structural data for the brain-wide morphology of neuronal and vascular networks. We found that a third of the imaged volume of the brain was affected by strong striated image intensity inhomogeneity and, furthermore, a significant amount of information content lost with Gaussian illumination was accessible when interrogated with Bessel beams. In conclusion, Bessel beams produce high-fidelity structural data of improved image homogeneity and might significantly relax demands placed on the automated tools to count, trace, or segment fluorescent features of interest

High-fidelity imaging in brain-wide structural studies using light-sheet microscopy / Müllenbroich, M. Caroline; Silvestri, Ludovico; Di Giovanna, Antonino P.; Mazzamuto, Giacomo; Costantini, Irene; Sacconi, Leonardo; Pavone, Francesco S.. - In: ENEURO. - ISSN 2373-2822. - STAMPA. - 5:(2018), pp. ENEURO.0124-18.2018-ENEURO.0124-18.2018. [10.1523/ENEURO.0124-18.2018]

High-fidelity imaging in brain-wide structural studies using light-sheet microscopy

Silvestri, Ludovico;DI GIOVANNA, ANTONINO PAOLO;Mazzamuto, Giacomo;Costantini, Irene;Sacconi, Leonardo;Pavone, Francesco S.
2018

Abstract

Light-sheet microscopy (LSM) has proven a useful tool in neuroscience to image whole brains with high frame rates at cellular resolution and, in combination with tissue clearing methods, is often employed to reconstruct the cyto-architecture over the intact mouse brain. Inherently to LSM, however, residual opaque objects, always present to some extent even in extremely well optically cleared samples, cause stripe artifacts, which, in the best case, severely affect image homogeneity and, in the worst case, completely obscure features of interest. Here, demonstrating two example applications in intact optically cleared mouse brains, we report how Bessel beams reduce streaking artifacts and produce high-fidelity structural data for the brain-wide morphology of neuronal and vascular networks. We found that a third of the imaged volume of the brain was affected by strong striated image intensity inhomogeneity and, furthermore, a significant amount of information content lost with Gaussian illumination was accessible when interrogated with Bessel beams. In conclusion, Bessel beams produce high-fidelity structural data of improved image homogeneity and might significantly relax demands placed on the automated tools to count, trace, or segment fluorescent features of interest
2018
5
ENEURO.0124-18.2018
ENEURO.0124-18.2018
Goal 3: Good health and well-being for people
Goal 9: Industry, Innovation, and Infrastructure
Müllenbroich, M. Caroline; Silvestri, Ludovico; Di Giovanna, Antonino P.; Mazzamuto, Giacomo; Costantini, Irene; Sacconi, Leonardo; Pavone, Francesco ...espandi
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Utilizza questo identificatore per citare o creare un link a questa risorsa: https://hdl.handle.net/2158/1152070
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