A protocol for extracting immunolabeled murine cardiomyocytes of high-quality RNA by laser capture microdissection

Abeer K. Shaalan, Georgina M. Ellison-Hughes*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

1 Citation (Scopus)

Abstract

We developed a highly efficient, ultrashort immunohistochemistry-laser capture microdissection (IHC-LMD) protocol, which allows microdissection of up to 250 single cardiomyocytes. Before LMD, murine hearts are excised, snap-frozen, and cryosectioned. RNA isolated from LMD material is of high RNA quality, making it usable for gene expression analysis and RNA sequencing. Challenges and limitations of this protocol include visualization of the immunostaining and nuclei DAPI dye on the PEN slides, and timing and speed to limit RNA degradation as much as possible.

Original languageEnglish
Article number101231
JournalSTAR Protocols
Volume3
Issue number1
Early online date8 Mar 2022
DOIs
Publication statusPublished - 18 Mar 2022

Keywords

  • Cell isolation
  • Gene Expression
  • Microscopy
  • Molecular Biology

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