title: Assessment of beta cell viability.
abstract: This unit contains detailed protocols for the simultaneous identification of the human pancreatic β cells and determination of their viability by flow cytometry. The enumeration of β cells is based on the ability of the cell-permeable form of the zinc-selective dye, FluoZin-3-AM, to bind intracellular labile zinc stored at higher levels in these cells than any other types of cells in the body. Although staining of intracellular labile zinc by FluoZin-3-AM is dependent on the metabolic activity of β cells, co-staining with a mitochondrial transmembrane potential indicator allows the accurate determination of viability. Simultaneous measurement of intracellular antioxidant thiols is also compatible with the detection of β cells containing metabolically active mitochondria. The method for assessing the mitochondrial functionality by flow cytometry described herein is simple to perform and sufficient to detect the viability of β cells in human islet preparations.
authors.affiliation: University of Illinois at Chicago, Chicago, Illinois.
authors.forename: Sundararajan
authors.initials: S
authors.lastname: Jayaraman
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D002470:Cell Survival; D005434:Flow Cytometry / Q000295:instrumentation / Q000379:methods*; D005456:Fluorescent Dyes / Q000494:pharmacology; D006801:Humans; D050417:Insulin-Secreting Cells / Q000166:cytology* / Q000502:physiology*; D008954:Models, Biological; D013194:Staining and Labeling / Q000379:methods
chemical_list: D005456:Fluorescent Dyes