Flow cytometric analysis of DNA and nuclear protein in paraffin‐embedded tissue

Gaetano Ciancio, Alan Pollack, Norman L. Block

Research output: Contribution to journalArticle

11 Scopus citations

Abstract

Previously we described the simultaneous quantification of DNA and nuclear protein in unfixed tissue from solid tumors. The resultant 2 parameter flow cytometric analysis has several advantages over that of DNA alone. In this report, we describe a modification of the technique for the analysis of formalin-fixed paraffin-embedded tissue. Paraffin-embedded material was prepared by hydrating sections, incubating in 0.5% pepsin solution, washing, and resuspending in buffer containing nonionic detergent. The nuclei were then stained with fluorescein isothiocyanate and propidium iodide in the presence of ribonuclease. Several solid tumor tissue types have been analyzed, including breast, colon, kidney, and thymus. The best results were obtained when the initial pepsin treatment was for 1.5 h, instead of 0.5 h. Pepsin treatment for 1.5 h improved the CVs of both the DNA and nuclear protein parameters, and did not appear to reduce the nuclear protein levels or to cause significant disintegration of nuclei. The DNA/nuclear protein histograms of unfixed and fixed, paraffin-embedded tissue was similar. Since tumor nuclei typically have higher protein levels than DNA-diploid nuclei, the technique reduces population overlapping and permits less subjective identification of DNA aneuploidy.

Original languageEnglish (US)
Pages (from-to)205-209
Number of pages5
JournalCytometry
Volume14
Issue number2
DOIs
StatePublished - 1993

Keywords

  • DNA
  • Flow cytometry
  • Nuclear protein
  • Paraffin-embedded

ASJC Scopus subject areas

  • Hematology
  • Cell Biology
  • Pathology and Forensic Medicine
  • Biophysics
  • Endocrinology

Fingerprint Dive into the research topics of 'Flow cytometric analysis of DNA and nuclear protein in paraffin‐embedded tissue'. Together they form a unique fingerprint.

  • Cite this