Journal
JOURNAL OF BIOMEDICAL OPTICS
Volume 15, Issue 1, Pages -Publisher
SPIE-SOC PHOTO-OPTICAL INSTRUMENTATION ENGINEERS
DOI: 10.1117/1.3324890
Keywords
structural imaging; 3-D imaging; macroscopy; light-sheet illumination
Funding
- NATIONAL INSTITUTE OF BIOMEDICAL IMAGING AND BIOENGINEERING [R21EB007338, R01EB010059] Funding Source: NIH RePORTER
- NIBIB NIH HHS [R01 EB010059-01, R21 EB007338, R01 EB010059, R21 EB007338-02] Funding Source: Medline
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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. (C) 2010 Society of Photo-Optical Instrumentation Engineers. [DOI: 10.1117/1.3324890]
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