Protocol for RNA fluorescence in situ hybridization in mouse meningeal whole mounts

Olof Rickard Nilsson; Laszlo Kari; Rebecca Rosenke; Olivia Steele-Mortimer

doi:10.1016/j.xpro.2022.101256

Protocol for RNA fluorescence in situ hybridization in mouse meningeal whole mounts

STAR Protoc. 2022 Mar 22;3(2):101256. doi: 10.1016/j.xpro.2022.101256. eCollection 2022 Jun 17.

Authors

Olof Rickard Nilsson¹, Laszlo Kari¹, Rebecca Rosenke², Olivia Steele-Mortimer¹

Affiliations

¹ Laboratory of Bacteriology, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, MT 59840-2932, USA.
² Rocky Mountain Veterinary Branch, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, MT 59840-2932, USA.

Abstract

The multilayered meninges surrounding the brain and spinal cord harbor distinct immune cell populations with prominent roles in health and diseases. Here we present an optimized protocol for RNA fluorescence in situ hybridization (RNA FISH) in meningeal whole mounts, allowing the visualization of gene expression. We also describe the combination of this protocol with immunohistochemistry for simultaneous visualization of mRNA and proteins. This protocol can be used for assessing spatial gene expression within the meninges.

Keywords: Antibody; Immunology; In Situ Hybridization; Microscopy; Neuroscience.

Publication types

Research Support, N.I.H., Intramural

MeSH terms

Animals
Brain*
Immunohistochemistry
In Situ Hybridization, Fluorescence / methods
Mice
RNA*
RNA, Messenger / genetics

Substances

RNA, Messenger
RNA