Scientific Results

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

Year: 2018

Authors: Müllenbroich M.C., Silvestri L., Di Giovanna A.P., Mazzamuto G., Costantini I., Sacconi L., Pavone F.S.

Autors Affiliation: National Institute of Optics, National Research Council, Sesto Fiorentino, 50019, Italy; European Laboratory for Non-Linear Spectroscopy, LENS, Sesto Fiorentino, 50019, Italy; University of Florence, Italy; Department of Physics and Astronomy, University of Florence, Sesto Fiorentino, 50019, Italy

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.

Journal/Review: ENEURO

Volume: 5 (6)      Pages from: e0124-1  to: e0124 -13

More Information: This project was supported by European Union’s H2020 Research and Innovation Programme Grants 720270 (Human Brain Project) and 654148 (Laserlab-Europe) and by the European Union’s Programme H2020 EXCEL-LENT SCIENCE-European Research Council (ERC) Grant 692943 (BrainBIT). The project has also been supported by the Italian Ministry for Education, University, and Research in the framework of the Flagship Project NanoMAX of Eurobioimaging Italian Nodes (ESFRI research infrastructure), and by “Ente Cassa di Risparmio di Firenze” (private foundation). Müllenbroich’s present address: School of Physics, Kelvin Building, University of Glasgow, G12 8QQ Glasgow, Scotland, United Kingdom. Acknowledgements: We thank Riccardo Ballerini and Ahmed Hajeb from the mechanical workshop at LENS for the production of custom pieces and advise on structural stability in sample mounting.
KeyWords: Animal experiment; article; artifact; brain; illumination; microscopy; morphology; mouse; nerve cell network; nonhuman
DOI: 10.1523/ENEURO.0124-18.2018

Citations: 5
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