Analysis of epicardial genes in embryonic mouse hearts with flow cytometry

STAR Protoc. 2021 Feb 24;2(1):100359. doi: 10.1016/j.xpro.2021.100359. eCollection 2021 Mar 19.

Abstract

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).

Keywords: Antibody; Cell isolation; Flow cytometry/mass cytometry; Gene expression; In situ hybridization.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Embryo, Mammalian / cytology
  • Embryo, Mammalian / metabolism*
  • Flow Cytometry*
  • Gene Expression Regulation, Developmental*
  • Mice
  • Pericardium / cytology
  • Pericardium / metabolism*