Unit

UNIT 7.15 Ultraviolet-Induced Detection of Halogenated Pyrimidines (UVID)

  1. Hans-Joerg Hammers,
  2. Peter Schlenke

Published Online: 1 MAY 2001

DOI: 10.1002/0471142956.cy0715s16

Current Protocols in Cytometry

Current Protocols in Cytometry

How to Cite

Hammers, H.-J. and Schlenke, P. 2001. Ultraviolet-Induced Detection of Halogenated Pyrimidines (UVID). Current Protocols in Cytometry. 16:7.15:7.15.1–7.15.6.

Author Information

  1. University of Luebeck, Leubeck, Germany

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: APR 2001

Abstract

When halogenated pyrimidines such as BrdU are used to identify DNA-synthesizing cells, their detection with monoclonal antibodies requires DNA denaturation, which can lead to loss of cells and/or cell markers or antigens. The authors present an alternative method employing ultraviolet light to partially photolyze BrdU and induce extensive damage in the nuclei which have incorporated the pyrimidine. Under appropriate conditions the BrdU in the unfolding chromatin is detected by certain monoclonal antibodies. Since no denaturation step or enzymatic treatment is required, this method preserves cellular markers and avoids enzyme-specific artifacts. Directions are given for both coagulative (ethanol) and crosslinking (formaldehyde) fixation. Keywords: flow cytometry; BrdU; UV; photolysis; hypotonic; cell cycle; proliferation When halogenated pyrimidines such as BrdU are used to identify DNA-synthesizing cells, their detection with monoclonal antibod