RT Journal Article
SR Electronic
T1 High-Fidelity Imaging in Brain-Wide Structural Studies Using Light-Sheet Microscopy
JF eneuro
JO eNeuro
FD Society for Neuroscience
SP ENEURO.0124-18.2018
DO 10.1523/ENEURO.0124-18.2018
VO 5
IS 6
A1 Müllenbroich, M. Caroline
A1 Silvestri, Ludovico
A1 Di Giovanna, Antonino P.
A1 Mazzamuto, Giacomo
A1 Costantini, Irene
A1 Sacconi, Leonardo
A1 Pavone, Francesco S.
YR 2018
UL http://www.eneuro.org/content/5/6/ENEURO.0124-18.2018.abstract
AB 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.