Unit

UNIT 6.27 High-Resolution Multiparameter DNA Flow Cytometry for the Detection and Sorting of Tumor and Stromal Subpopulations from Paraffin-Embedded Tissues

  1. Willem E. Corver,
  2. Natalja T. ter Haar

Published Online: 1 OCT 2009

DOI: 10.1002/0471142956.cy0627s50

Current Protocols in Cytometry

Current Protocols in Cytometry

How to Cite

Corver, W. E. and ter Haar, N. T. 2009. High-Resolution Multiparameter DNA Flow Cytometry for the Detection and Sorting of Tumor and Stromal Subpopulations from Paraffin-Embedded Tissues. Current Protocols in Cytometry. 50:6.27:6.27.1–6.27.21.

Author Information

  1. Leiden University Medical Centre, Leiden, The Netherlands

Publication History

  1. Published Online: 1 OCT 2009
  2. Published Print: OCT 2009

This is not the most recent version of the article. View current version (1 JAN 2011)

Abstract

This unit contains a detailed protocol for the simultaneous flow cytometric measurement of tumor cells, stromal cells, and DNA content of formalin-fixed, paraffin-embedded (FFPE) tissues. The vimentin-positive stromal cell fraction can be used as an internal reference for DNA content assessments. This allows clear detection of keratin-positive tumor cells with a DNA index lower than 1.0 and of keratin-positive tumor cells with a DNA close to 1.0 in overall DNA aneuploid samples, thus improved DNA ploidy assessment in FFPE carcinomas. Furthermore, the protocol is useful for studying molecular genetic alterations and intratumor heterogeneity in archival FFPE samples. Keratin-positive tumor cell fractions can be flow-sorted for further molecular genetic analysis, while DNA from the sorted vimentin-positive stromal cells can serve as a reference when normal tissue of the patient is not available. Curr. Protoc. Cytom. 50:6.27.1-6.27.21. © 2009 by John Wiley & Sons, Inc.

Keywords:

  • flow cytometry;
  • FFPE tissues;
  • DNA content;
  • stromal cells;
  • carcinoma cells;
  • vimentin;
  • keratin;
  • sorting;
  • genomic alterations