Unit

UNIT 7.40 Zinc Fixation for Flow Cytometry Analysis of Intracellular and Surface Epitopes, DNA Content, and Cell Proliferation

  1. Rikke Christensen1,
  2. David M. Owens2,
  3. Anette Thomsen3,
  4. Søren Pedersen1,
  5. Uffe Birk Jensen1,3

Published Online: 1 JUL 2011

DOI: 10.1002/0471142956.cy0740s57

Current Protocols in Cytometry

Current Protocols in Cytometry

How to Cite

Christensen, R., Owens, D. M., Thomsen, A., Pedersen, S. and Jensen, U. B. 2011. Zinc Fixation for Flow Cytometry Analysis of Intracellular and Surface Epitopes, DNA Content, and Cell Proliferation. Current Protocols in Cytometry. 57:7.40:7.40.1–7.40.9.

Author Information

  1. 1

    Department of Clinical Genetics, Aarhus University Hospital, Aarhus, Denmark

  2. 2

    Departments of Dermatology and Pathology, College of Physicians and Surgeons, Columbia University, New York, New York

  3. 3

    Department of Human Genetics, Aarhus University, Aarhus, Denmark

Publication History

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

Abstract

Zinc salt-based fixation (ZBF) is a simple, cost-effective, and nonhazardous fixation method for cell suspensions that preserves all cellular structures and enables flow cytometric analysis of both surface and intracellular proteins, DNA content profiles, and pulse-labeling using the thymidine analog EdU in the same cell sample. ZBF performs equally well to formaldehyde in the preservation of surface epitope labeling and forward and side light scatter parameters, as measured by flow cytometry. DNA is maintained at high molecular weight, improving the quantification and allowing subsequent quantitative PCR analysis. Finally, ZBF treatment allows for long-term storage of labeled cells with little change in these parameters. Curr. Protoc. Cytom. 57:7.40.1-7.40.9. © 2011 by John Wiley & Sons, Inc.

Keywords:

  • DNA content;
  • DNA synthesis;
  • Click-iT assay;
  • surface and intracellular markers