Effect of sonication on paraffin-embedded tissue preparation for DNA flow cytometry

N. J. Gonchoroff, J. J. Ryan, T. K. Kimlinger, Thomas Elmer Witzig, P. R. Greipp, J. S. Meyer, J. A. Katzmann

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

Since the publication of paraffin block extraction procedures, flow cytometric analysis of DNA ploidy and S-phase of tumor specimens has been widely applied. DNA aneuploidy, DNA tetraploid (elevated G2/M), and elevated S-phase are clinically significant in some tumor systems. True DNA tetraploid cell lines will contain a large 4c population and perhaps an 8c population; samples with cell aggregates will also contain a 6c population. Microscopic examination of samples having a 6c peak revealed nuclei with adhering debris and doublets, triplets, and larger nuclear aggregates. After sonication, a uniform suspension of single nuclei without adherent debris was seen. In addition to reducing the percent of G2/M cells, sonication also reduced S-phase percent such that it was closer to the bromodeoxyuridine labeling index. The DNA ploidy classification of specimens was also compared pre- and post-sonication. Four of 96 breast cancer samples changed classification; all were specimens in which the histogram became cleaner and small DNA aneuploid peak became apparent after sonication.

Original languageEnglish (US)
Pages (from-to)642-646
Number of pages5
JournalCytometry
Volume11
Issue number5
DOIs
StatePublished - 1990

Fingerprint

Sonication
Paraffin
Flow Cytometry
DNA
S Phase
Tetraploidy
Ploidies
Aneuploidy
Population
Bromodeoxyuridine
Publications
Neoplasms
Suspensions
Breast Neoplasms
Cell Line

Keywords

  • Breast cancer S-phase measurements
  • Cell clumping
  • DNA aneuploidy
  • Propidium iodide
  • Sample aggregation

ASJC Scopus subject areas

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

Cite this

Gonchoroff, N. J., Ryan, J. J., Kimlinger, T. K., Witzig, T. E., Greipp, P. R., Meyer, J. S., & Katzmann, J. A. (1990). Effect of sonication on paraffin-embedded tissue preparation for DNA flow cytometry. Cytometry, 11(5), 642-646. https://doi.org/10.1002/cyto.990110513

Effect of sonication on paraffin-embedded tissue preparation for DNA flow cytometry. / Gonchoroff, N. J.; Ryan, J. J.; Kimlinger, T. K.; Witzig, Thomas Elmer; Greipp, P. R.; Meyer, J. S.; Katzmann, J. A.

In: Cytometry, Vol. 11, No. 5, 1990, p. 642-646.

Research output: Contribution to journalArticle

Gonchoroff, NJ, Ryan, JJ, Kimlinger, TK, Witzig, TE, Greipp, PR, Meyer, JS & Katzmann, JA 1990, 'Effect of sonication on paraffin-embedded tissue preparation for DNA flow cytometry', Cytometry, vol. 11, no. 5, pp. 642-646. https://doi.org/10.1002/cyto.990110513
Gonchoroff, N. J. ; Ryan, J. J. ; Kimlinger, T. K. ; Witzig, Thomas Elmer ; Greipp, P. R. ; Meyer, J. S. ; Katzmann, J. A. / Effect of sonication on paraffin-embedded tissue preparation for DNA flow cytometry. In: Cytometry. 1990 ; Vol. 11, No. 5. pp. 642-646.
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