Protocol for multiplex fluorescent immunohistochemistry in free-floating rodent brain tissues

Aubrey M Kelly; Brandon A Fricker; Kelly J Wallace

doi:10.1016/j.xpro.2022.101672

Protocol for multiplex fluorescent immunohistochemistry in free-floating rodent brain tissues

STAR Protoc. 2022 Dec 16;3(4):101672. doi: 10.1016/j.xpro.2022.101672. Epub 2022 Sep 14.

Authors

Aubrey M Kelly¹, Brandon A Fricker², Kelly J Wallace²

Affiliations

¹ Department of Psychology, Emory University, 36 Eagle Row, Atlanta, GA 30322, USA. Electronic address: aubrey.kelly@emory.edu.
² Department of Psychology, Emory University, 36 Eagle Row, Atlanta, GA 30322, USA.

Abstract

Identifying multiple proteins within the same tissue allows for assessing protein colocalization, is cost effective, and maximizes efficiency. Here, we describe a protocol for multiplex immunolabeling of proteins in free-floating rodent brain sections. As opposed to slide-mounted immunohistochemistry, the free-floating approach results in less tissue loss and greater antibody penetration. Using distinct fluorophores for individual proteins, this protocol allows for visualization of three or more proteins within tissue sections. The protocol can be applied to other tissue types. For complete details on the use and execution of this protocol, please refer to Gonzalez Abreu et al. (2022).

Keywords: Antibody; Microscopy; Molecular Biology; Neuroscience.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Antibodies*
Brain
Fluorescent Dyes*
Immunohistochemistry

Substances

Fluorescent Dyes
Antibodies