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Journal of Biomedical Optics • Open Access

Scanning light-sheet microscopy in the whole mouse brain with HiLo background rejection
Author(s): Jerome Mertz; Jinhyun Kim

Paper Abstract

It is well known that light-sheet illumination can enable optically sectioned wide-field imaging of macroscopic samples. However, the optical sectioning capacity of a light-sheet macroscope is undermined by sample-induced scattering or aberrations that broaden the thickness of the sheet illumination. We present a technique to enhance the optical sectioning capacity of a scanning light-sheet microscope by out-of-focus background rejection. The technique, called HiLo microscopy, makes use of two images sequentially acquired with uniform and structured sheet illumination. An optically sectioned image is then synthesized by fusing high and low spatial frequency information from both images. The benefits of combining light-sheet macroscopy and HiLo background rejection are demonstrated in optically cleared whole mouse brain samples, using both green fluorescent protein (GFP)-fluorescence and dark-field scattered light contrast.

Paper Details

Date Published: 1 January 2010
PDF: 7 pages
J. Biomed. Opt. 15(1) 016027 doi: 10.1117/1.3324890
Published in: Journal of Biomedical Optics Volume 15, Issue 1
Show Author Affiliations
Jerome Mertz, Boston Univ. (United States)
Jinhyun Kim, Howard Hughes Medical Institute (United States)

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