Work

Spatial analysis of transcript and protein levels in skeletal muscle

Year: 2024

Type: article

Abstract: Skeletal muscle spatial analyses have revealed unexpected regionalized gene expression patterns challenging the understanding of muscle as a homogeneous tissue. Here, we present a protocol for the spatial analysis of transcript and protein levels in murine skeletal muscle. We describe steps for tibialis anterior dissection, formaldehyde fixation, tissue chopper cutting, and hybridization chain reaction (HCR) detection and amplification. We then detail procedures for immunostaining, tissue clearing, and imaging. This protocol is easily adaptable to other tissues. • Detailed protocol for spatial analysis of transcripts and proteins in murine TA muscle • Steps for obtaining optimal thick sections of fixed TA muscle • Guide for hybridization chain reaction (HCR) detection and amplification • Steps for immunostaining, clearing, and confocal imaging of thick muscle sections Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Skeletal muscle spatial analyses have revealed unexpected regionalized gene expression patterns challenging the understanding of muscle as a homogeneous tissue. Here, we present a protocol for the spatial analysis of transcript and protein levels in murine skeletal muscle. We describe steps for tibialis anterior dissection, formaldehyde fixation, tissue chopper cutting, and hybridization chain reaction (HCR) detection and amplification. We then detail procedures for immunostaining, tissue clearing, and imaging. This protocol is easily adaptable to other tissues.

Source: STAR Protocols

DOI: 10.1016/j.xpro.2024.103378