A novel Ly6C/Ly6G-based strategy to analyze the mouse splenic myeloid compartment

Shawn Rose, Alexander Misharin, Harris Perlman*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

264 Scopus citations

Abstract

Currently, there is no standardized panel for immunophenotyping myeloid cells in mouse spleen using flow cytometry. Markers such as CD11b, CD11c, F4/80, Gr-1, Ly6C, and Ly6G have long been used to identify various splenic cell myeloid populations. Flow cytometry and fluorescence-activated cell sorting (FACS) analysis demonstrated that Ly6G/Ly6C markers are superior to Gr-1 for identifying splenic neutrophils, eosinophils, and subsets of monocytes/macrophages. Moreover, these experiments showed that F4/80 is not required for identifying these myeloid subsets and that many of the commercially available preparations of anti-F4/80 antibodies stain poorly for this antigen in spleen. Taken together, we have now developed an informative flow cytometry panel that can be combined with other cell markers to further delineate subpopulations of mouse splenic myeloid cells. This panel will be highly useful to investigators in the flow cytometry field, as there is a critical need to standardize the analysis of myeloid cell subsets.

Original languageEnglish (US)
Pages (from-to)343-350
Number of pages8
JournalCytometry Part A
Volume81 A
Issue number4
DOIs
StatePublished - Apr 2012

Keywords

  • F4/80
  • Immunophenotyping
  • Macrophage
  • Monocyte

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Cell Biology
  • Histology

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