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Small Methods, First published: 05 November 2021 | https://doi.org/10.1002/smtd.202100943 Sodium Cholate-Based Active Delipidation for Rapid and Efficient Clearing and Immunostaining of Deep Biological Samples

Myeongsu Na, Kitae Kim, Kyoungjoon Oh, Hyung Jin Choi, Chang Man Ha, and Sunghoe Chang^*

M. Na, K. Kim, K. Oh, S. Chang
Department of Physiology and Biomedical Sciences Seoul National University College of Medicine Seoul 03080, South Korea

H. J. Choi
Department of Anatomy and Biomedical Sciences Seoul National University College of Medicine Seoul 03080, South Korea

C. M. Ha
Research Division and Brain Research Core Facility Korea Brain Research Institute Daegu 41068, South Korea

S. Chang
Neuroscience Research Institute Seoul National University College of Medicine Seoul 03080, South Korea

^*Corresponding author.

Abstract

Recent surges of optical clearing provided anatomical maps to understand structure-function relationships at organ scale. Detergent-mediated lipid removal enhances optical clearing and allows efficient penetration of antibodies inside tissues, and sodium dodecyl sulfate (SDS) is the most common choice for this purpose. SDS, however, forms large micelles and has a low critical micelle concentration (CMC). Theoretically, detergents that form smaller micelles and higher CMC should perform better but these have remained mostly unexplored. Here, SCARF, a sodium cholate (SC)-based active delipidation method, is developed for better clearing and immunolabeling of thick tissues or whole organs. It is found that SC has superior properties to SDS as a detergent but has serious problems; precipitation and browning. These limitations are overcome by using the ion-conductive film to confine SC while enabling high conductivity. SCARF renders orders of magnitude faster tissue transparency than the SDS-based method, while excellently preserving the endogenous fluorescence, and enables much efficient penetration of a range of antibodies, thus revealing structural details of various organs including sturdy post-mortem human brain tissues at the cellular resolution. Thus, SCARF represents a robust and superior alternative to the SDS-based clearing methods and is expected to facilitate the 3D morphological mapping of various organs.

논문정보

형식Research article
게재일2021년 11월 (BRIC 등록일 2021-11-11)
연구진국내 연구진
분야 의약학 > 신경의학

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