Comprehensive optical and data management infrastructure for high-throughput light-sheet microscopy of whole mouse brains

M. Caroline Müllenbroich; Ludovico Silvestri; Leonardo Onofri; Irene Costantini; Marcel van’t Hoff; Leonardo Sacconi; Giulio Iannello; Francesco S. Pavone

doi:10.1117/1.NPh.2.4.041404

29 June 2015 Comprehensive optical and data management infrastructure for high-throughput light-sheet microscopy of whole mouse brains

M. Caroline Müllenbroich, Ludovico Silvestri, Leonardo Onofri, Irene Costantini, Marcel van’t Hoff, Leonardo Sacconi, Giulio Iannello, Francesco S. Pavone

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Neurophotonics, Vol. 2, Issue 4, 041404 (June 2015). https://doi.org/10.1117/1.NPh.2.4.041404

Abstract

Comprehensive mapping and quantification of neuronal projections in the central nervous system requires high-throughput imaging of large volumes with microscopic resolution. To this end, we have developed a confocal light-sheet microscope that has been optimized for three-dimensional (3-D) imaging of structurally intact clarified whole-mount mouse brains. We describe the optical and electromechanical arrangement of the microscope and give details on the organization of the microscope management software. The software orchestrates all components of the microscope, coordinates critical timing and synchronization, and has been written in a versatile and modular structure using the LabVIEW language. It can easily be adapted and integrated to other microscope systems and has been made freely available to the light-sheet community. The tremendous amount of data routinely generated by light-sheet microscopy further requires novel strategies for data handling and storage. To complete the full imaging pipeline of our high-throughput microscope, we further elaborate on big data management from streaming of raw images up to stitching of 3-D datasets. The mesoscale neuroanatomy imaged at micron-scale resolution in those datasets allows characterization and quantification of neuronal projections in unsectioned mouse brains.

CC BY: © The Authors. Published by SPIE under a Creative Commons Attribution 4.0 Unported License. Distribution or reproduction of this work in whole or in part requires full attribution of the original publication, including its DOI.

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M. Caroline Müllenbroich, Ludovico Silvestri, Leonardo Onofri, Irene Costantini, Marcel van’t Hoff, Leonardo Sacconi, Giulio Iannello, and Francesco S. Pavone "Comprehensive optical and data management infrastructure for high-throughput light-sheet microscopy of whole mouse brains," Neurophotonics 2(4), 041404 (29 June 2015). https://doi.org/10.1117/1.NPh.2.4.041404

Published: 29 June 2015

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