Unit

UNIT 7.31 Assessment of Cell Proliferation by 5-Bromodeoxyuridine (BrdU) Labeling for Multicolor Flow Cytometry

  1. Kristina Rothaeusler,
  2. Nicole Baumgarth

Published Online: 1 APR 2007

DOI: 10.1002/0471142956.cy0731s40

Current Protocols in Cytometry

Current Protocols in Cytometry

How to Cite

Rothaeusler, K. and Baumgarth, N. 2007. Assessment of Cell Proliferation by 5-Bromodeoxyuridine (BrdU) Labeling for Multicolor Flow Cytometry. Current Protocols in Cytometry. 40:7.31:7.31.1–7.31.13.

Author Information

  1. University of California, Davis, Davis, California

Publication History

  1. Published Online: 1 APR 2007
  2. Published Print: APR 2007

Abstract

Cell proliferation assays are used for a large variety of applications in the life sciences. This unit describes a flow-cytometry-based method that uses BrdU labeling in conjunction with multiple fluorescently labeled cell surface markers, allowing extensive phenotypic characterization of dividing cells in addition to assessment of their frequency. BrdU labeling has the advantage of constituting a nonradioactive technique that, when combined with additional fluorescent-based reagents, avoids time-consuming and often costly cell isolation procedures. Moreover, because BrdU is stably integrated into the DNA, it can be measured in a cell for many months. The method presented in this unit is based on paraformaldehyde fixation and reversible saponin-based cell membrane permeabilization, which maintains cell integrity as well as fluorescent labeling with a large number of different fluorochromes, allowing 10- to 12-color flow cytometric analysis of proliferating cells.

Keywords:

  • DNA labeling;
  • intracellular staining;
  • FACS;
  • turnover;
  • proliferation