Unit

UNIT 9.26 Immunophenotyping and DNA Content Analysis of Acetone-Fixed Cells

  1. Debora Mancaniello,
  2. Maurizio Carbonari

Published Online: 1 OCT 2008

DOI: 10.1002/0471142956.cy0926s46

Current Protocols in Cytometry

Current Protocols in Cytometry

How to Cite

Mancaniello, D. and Carbonari, M. 2008. Immunophenotyping and DNA Content Analysis of Acetone-Fixed Cells. Current Protocols in Cytometry. 46:9.26:9.26.1–9.26.11.

Author Information

  1. Laboratory of Immunology, Clinical Medicine Department, University of Rome “La Sapienza,”, Rome, Italy

Publication History

  1. Published Online: 1 OCT 2008
  2. Published Print: OCT 2008

Abstract

The flow acetone staining technique (FAST) allows one to concurrently study physical cell features revealed by light-scatter analysis, surface/nuclear phenotypes, and cellular DNA content. Thus, diverse subpopulations of proliferating cells can be identified in heterogeneous populations by their immunophenotype and their cell cycle status, and DNA ploidy can be assessed. Acetone, a coagulant (precipitating) fixative that also has the ability to permeabilize cell membranes, is widely used in static cytometry, but rarely in flow cytometry because of its undesirable effects, namely causing cell shrinkage. Nevertheless, when employed under proper temperature conditions (∼8°C), it preserves cellular physical features and immunophenotype well, and is compatible with stoichiometric DNA staining and accurate measurement of DNA content. Due to these virtues of FAST, the method provides useful approaches for cell biology and hematology/oncology studies. Curr. Protocol. Cytom. 46:9.26.1-9.26.11. © 2008 by John Wiley & Sons, Inc.

Keywords:

  • acetone;
  • multiparametric flow cytometry;
  • DNA content;
  • conservative fixation;
  • proliferating cells