Unit

UNIT 6.27 Assessment of Beta Cell Viability

  1. Sundararajan Jayaraman

Published Online: 1 JAN 2011

DOI: 10.1002/0471142956.cy0627s55

Current Protocols in Cytometry

Current Protocols in Cytometry

How to Cite

Jayaraman, S. 2011. Assessment of Beta Cell Viability. Current Protocols in Cytometry. 55:6.27:6.27.1–6.27.16.

Author Information

  1. University of Illinois at Chicago, Chicago, Illinois

Publication History

  1. Published Online: 1 JAN 2011
  2. Published Print: JAN 2011

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. Curr. Protoc. Cytom. 55:6.27.1-6.27.16. © 2010 by John Wiley & Sons, Inc.

Keywords:

  • beta cells;
  • flow cytometry;
  • glutathione;
  • labile zinc;
  • mitochondrial function;
  • thiols