Check samples for laboratory self-assessment in DNA flow cytometry. The National Cancer Institute's Flow Cytometry Network experience

J. S. Coon, A. D. Deitch, R. W. De Vere White, L. G. Koss, M. R. Melamed, J. E. Reeder, Ronald S Weinstein, R. P. Wersto, L. L. Wheeless

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

The National Cancer Institute's Flow Cytometry Network (NCI-FCN) is attempting to facilitate the transfer of flow cytometry (FCM) of exfoliated bladder cells from the research laboratory to the clinical laboratory. Demonstrating interinstitutional consistency in FCM analysis of replicate specimens simulating clinical barbotage specimens, fixed to allow easy transportation and storage at room temperature was one specific objective. Simulated barbotage specimens were prepared by mixing cultured aneuploid bladder carcinoma cells with normal or mitogen-stimulated peripheral blood mononuclear cells in different ratios. The samples were fixed in 10% formalin for 30 minutes, stored in buffer, and enucleated with pepsin, pH 1.5, before staining with propidium iodide for FCM DNA analysis. Preservation in ethanol or other common DNA cytochemical reagents was found to be unsatisfactory. In contrast, the formalin-fixed samples showed excellent preservation of quantitative DNA fluorescence and coefficient of variation of histogram peaks for over 2 weeks. Exchange of eight fixed specimens among five network laboratories that analyzed them as 'unknowns' showed good overall agreement on histogram data and interpretation, although some noteworthy interlaboratory differences were found. This technique could be used for self-assessment surveys of clinical laboratory performance in DNA FCM of bladder barbotage specimens.

Original languageEnglish (US)
Pages (from-to)1592-1599
Number of pages8
JournalCancer
Volume63
Issue number8
StatePublished - 1989
Externally publishedYes

Fingerprint

National Cancer Institute (U.S.)
Flow Cytometry
DNA
Urinary Bladder
Formaldehyde
Propidium
Pepsin A
Aneuploidy
Mitogens
Blood Cells
Buffers
Ethanol
Fluorescence
Self-Assessment
Staining and Labeling
Carcinoma
Temperature
Research

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Coon, J. S., Deitch, A. D., De Vere White, R. W., Koss, L. G., Melamed, M. R., Reeder, J. E., ... Wheeless, L. L. (1989). Check samples for laboratory self-assessment in DNA flow cytometry. The National Cancer Institute's Flow Cytometry Network experience. Cancer, 63(8), 1592-1599.

Check samples for laboratory self-assessment in DNA flow cytometry. The National Cancer Institute's Flow Cytometry Network experience. / Coon, J. S.; Deitch, A. D.; De Vere White, R. W.; Koss, L. G.; Melamed, M. R.; Reeder, J. E.; Weinstein, Ronald S; Wersto, R. P.; Wheeless, L. L.

In: Cancer, Vol. 63, No. 8, 1989, p. 1592-1599.

Research output: Contribution to journalArticle

Coon, JS, Deitch, AD, De Vere White, RW, Koss, LG, Melamed, MR, Reeder, JE, Weinstein, RS, Wersto, RP & Wheeless, LL 1989, 'Check samples for laboratory self-assessment in DNA flow cytometry. The National Cancer Institute's Flow Cytometry Network experience', Cancer, vol. 63, no. 8, pp. 1592-1599.
Coon JS, Deitch AD, De Vere White RW, Koss LG, Melamed MR, Reeder JE et al. Check samples for laboratory self-assessment in DNA flow cytometry. The National Cancer Institute's Flow Cytometry Network experience. Cancer. 1989;63(8):1592-1599.
Coon, J. S. ; Deitch, A. D. ; De Vere White, R. W. ; Koss, L. G. ; Melamed, M. R. ; Reeder, J. E. ; Weinstein, Ronald S ; Wersto, R. P. ; Wheeless, L. L. / Check samples for laboratory self-assessment in DNA flow cytometry. The National Cancer Institute's Flow Cytometry Network experience. In: Cancer. 1989 ; Vol. 63, No. 8. pp. 1592-1599.
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