Preparation of a Single Cell Suspension from the Murine Iridocorneal Angle

Benjamin Robert Thomson*, Susan E. Quaggin

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

Single cell RNA sequencing is a powerful tool that can be used to identify distinct cell types and transcriptomic differences within complex tissues. It has proven to be especially useful in tissues of the eye, where investigators have identified novel cell types within the retina, anterior chamber, and iridocorneal angle and explored transcriptomic contribution to disease phenotypes in age-related macular degeneration. However, to obtain high quality results, the technique requires isolation of healthy single cells from the tissue of interest, seeking complete tissue digestion while minimizing stress and transcriptomic changes in the isolated cells prior to library preparation. Here, we present a protocol developed in our laboratory for isolation of live single cells from the murine iridocorneal angle, which includes Schlemm’s canal and the trabecular meshwork, suitable for single cell RNA sequencing, flow cytometry, or other downstream analysis.

Original languageEnglish (US)
Article numbere4426
JournalBio-protocol
Volume12
Issue number10
DOIs
StatePublished - May 20 2022

Keywords

  • Dissociation
  • Eye
  • Glaucoma
  • Iridocorneal angle
  • Limbus
  • Mouse
  • Schlemm’s canal
  • Single cell
  • Single cell RNA sequencing

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)
  • Neuroscience(all)
  • Plant Science

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