Measurement variability in DNA flow cytometry of replicate samples

Leon L. Wheeless, John S. Coon, Christopher Cox, Arline D. Deitch, Ralph W. Devere White, Leopold G. Koss, Myron R. Melamed, Mary J. O'Connell, Jay E. Reeder, Ronald S. Weinstein, Robert P. Wersto

Research output: Contribution to journalArticle

29 Scopus citations

Abstract

A Bladder Cancer Flow Cytometry Network study has been carried out aimed at identification of the sources of inter‐ and intralaboratory variability. Replicate “cocktail” samples containing a mixture of peripheral blood lymphocytes and an aneuploid cell line and samples of peripheral blood lymphocytes serving as a DNA reference standard were distributed to five network laboratories. The samples were stained for DNA using propidium iodide, with each laboratory using its own staining protocol. Sets of these samples were analyzed by flow cytometry to obtain cellular DNA distributions. DNA index and hyperdiploid fraction were calculated for each histogram using an automated technique. Results were evaluated by analysis of variance to identify sources of variability. Three important sources of variation were found that affect flow cytometry in general and‐the transportability of flow cytometry results to routine clinical use in particular. The significant variation among laboratories that is constant across time most probably represents stable differences in instrumentation, instrument set‐up, and laboratory techniques. This variation can be compensated for, if it is known and stable, to develop transportable classification criteria. The second type of variation, termed the interaction component, represents differences among laboratories that are not constant across time. Sources of this variation include inconsistency in sample preparation, staining, and analysis. The elimination of this type of variation is required for meaningful comparison of data within and among laboratories and the creation of interlaboratory databases. The third type of variation represents pure measurement variability and affects the sensitivity of the technique.

Original languageEnglish (US)
Pages (from-to)731-738
Number of pages8
JournalCytometry
Volume10
Issue number6
DOIs
StatePublished - Nov 1989
Externally publishedYes

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Keywords

  • Measurement variability
  • laboratory variability
  • quality control

ASJC Scopus subject areas

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

Cite this

Wheeless, L. L., Coon, J. S., Cox, C., Deitch, A. D., Devere White, R. W., Koss, L. G., Melamed, M. R., O'Connell, M. J., Reeder, J. E., Weinstein, R. S., & Wersto, R. P. (1989). Measurement variability in DNA flow cytometry of replicate samples. Cytometry, 10(6), 731-738. https://doi.org/10.1002/cyto.990100610