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Low cost light-sheet microscopy for whole brain imaging
Author(s): Manish Kumar; Jordan Nasenbeny ; Yevgenia Kozorovitskiy
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Paper Abstract

Light-sheet microscopy has evolved as an indispensable tool in imaging biological samples. It can image 3D samples at fast speed, with high-resolution optical sectioning, and with reduced photobleaching effects. These properties make light-sheet microscopy ideal for imaging fluorophores in a variety of biological samples and organisms, e.g. zebrafish, drosophila, cleared mouse brains, etc. While most commercial turnkey light-sheet systems are expensive, the existing lower cost implementations, e.g. OpenSPIM, are focused on achieving high-resolution imaging of small samples or organisms like zebrafish. In this work, we substantially reduce the cost of light-sheet microscope system while targeting to image much larger samples, i.e. cleared mouse brains, at single-cell resolution. The expensive components of a lightsheet system – excitation laser, water-immersion objectives, and translation stage – are replaced with an incoherent laser diode, dry objectives, and a custom-built Arduino-controlled translation stage. A low-cost CUBIC protocol is used to clear fixed mouse brain samples. The open-source platforms of μManager and Fiji support image acquisition, processing, and visualization. Our system can easily be extended to multi-color light-sheet microscopy.

Paper Details

Date Published: 23 February 2018
PDF: 6 pages
Proc. SPIE 10499, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXV, 104991I (23 February 2018); doi: 10.1117/12.2288497
Show Author Affiliations
Manish Kumar, Northwestern Univ. (United States)
Jordan Nasenbeny , Northwestern Univ. (United States)
Yevgenia Kozorovitskiy, Northwestern Univ. (United States)

Published in SPIE Proceedings Vol. 10499:
Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXV
Thomas G. Brown; Carol J. Cogswell; Tony Wilson, Editor(s)

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