Analysis of epicardial genes in embryonic mouse hearts with flow cytometry

Highlights

• A protocol to characterize co-expression of genes in the epicardium and its derivatives

• Dissociation of embryonic mouse hearts into a reliable single-cell suspension

• Multiplexed RNA-ISH staining paired with cell lineage- and immuno-labeling

• Single-cell resolution analysis of epicardial marker expression by flow cytometry

Summary

Genetic markers used to define discrete cell populations are seldom expressed exclusively in the population of interest and are, thus, unsuitable when evaluated individually, especially in the absence of spatial and morphological information. Here, we present fluorescence in situ hybridization for flow cytometry to allow simultaneous analysis of multiple marker genes at the single whole-cell level, exemplified by application to the embryonic epicardium. The protocol facilitates multiplexed quantification of gene and protein expression and temporal changes across specific cell populations.

For complete details on the use and execution of this protocol, please refer to Lupu et al. (2020).