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Journal of Biomedical Optics • Open Access

Quantification of light attenuation in optically cleared mouse brains
Author(s): Angela d’Esposito; Daniil Nikitichev; Adrien Desjardins; Simon Walker-Samuel; Mark F. Lythgoe

Paper Abstract

Optical clearing, in combination with recently developed optical imaging techniques, enables visualization and acquisition of high-resolution, three-dimensional images of biological structures deep within the tissue. Many different approaches can be used to reduce light absorption and scattering within the tissue, but there is a paucity of research on the quantification of clearing efficacy. With the use of a custom-made spectroscopy system, we developed a way to quantify the quality of clearing in biological tissue and applied it to the mouse brain. Three clearing techniques were compared: BABB (1:2 mixture of benzyl alcohol and benzyl benzoate, also known as Murray’s clear), pBABB (peroxide BABB, a modification of BABB which includes the use of hydrogen peroxide), and passive CLARITY. We found that BABB and pBABB produced the highest degree of optical clearing. Furthermore, the approach allows regional measurement of light attenuation to be performed, and our results show that light is most attenuated in regions with high lipid content. We provide a way to choose between the multiple clearing protocols available, and it could prove useful for evaluating images that are acquired with cleared tissues.

Paper Details

Date Published: 17 August 2015
PDF: 3 pages
J. Biomed. Opt. 20(8) 080503 doi: 10.1117/1.JBO.20.8.080503
Published in: Journal of Biomedical Optics Volume 20, Issue 8
Show Author Affiliations
Angela d’Esposito, Univ. College London (United Kingdom)
Daniil Nikitichev, Univ. College London (United Kingdom)
Adrien Desjardins, Univ. College London (United Kingdom)
Simon Walker-Samuel, Univ. College London (United Kingdom)
Mark F. Lythgoe, Univ. College London (United Kingdom)

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