Technical Advance: Autofluorescence as a tool for myeloid cell analysis

Journal of Leukocyte Biology

Nicholas H. Hunt²  Georges E. Grau²  Nico Van Rooijen¹  Giovanna Chimini³  Lionel Chasson³  Andrew J. Mitchell³  Lydie C. Pradel³ 

[1] Department of Molecular Cell Biology, Vrije Universiteit, VUMC, Amsterdam, The Netherlands Department of Molecular Cell Biology, Vrije Universiteit, VUMC, Amsterdam, The Netherlands Department of Molecular Cell Biology, Vrije Universiteit, VUMC, Amsterdam, The Netherlands;Bosch Institute and School of Medical Sciences, University of Sydney, New South Wales, Australia; and Bosch Institute and School of Medical Sciences, University of Sydney, New South Wales, Australia; and Bosch Institute and School of Medical Sciences, University of Sydney, New South Wales, Australia; andCentre d’Immunologie de Marseille-Luminy, INSERM-CNRS-Université de La Méditerranée, Marseille, France; Centre d’Immunologie de Marseille-Luminy, INSERM-CNRS-Université de La Méditerranée, Marseille, France; Centre d’Immunologie de Marseille-Luminy, INSERM-CNRS-Université de La Méditerranée, Marseille, France;

关键词: macrophage;    monocyte;    flow cytometry;    spleen;   

DOI  :  10.1189/jlb.0310184

学科分类：生理学

来源: Federation of American Societies for Experimental Biology

【 摘 要 】

Cellular AF is usually considered a hindrance to flow cytometric analysis. Here, we incorporate AF into analysis of complex mixtures of leukocytes. Using a mouse model, we examined cellular AF at multiple excitation and emission wavelengths, and populations with discrete patterns were gated and examined for surface marker expression. In the spleen, all major myeloid populations were identified. In particular, the approach allowed simultaneous characterization of RPM and resident monocytes. When monocytes and RPM were compared, RPM exhibited a phenotype that was consistent with involvement in physiological processes, including expression of genes involved in lipid and iron metabolism. The presence of large amounts of stored ferric iron within RPM enabled purification of these cells using a magnetic-based approach. When adapted for use on leukocytes isolated from a range of other organs, incorporation of AF into analysis allowed identification and isolation of biologically important myeloid populations, including subsets that were not readily identifiable by conventional cytometric analysis.

【 授权许可】

Unknown