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Check samples for laboratory self‐assessment in DNA flow cytometry. The national cancer institute's flow cytometry network experience
Author(s) -
Coon John S.,
Deitch Arline D.,
de Vere White Ralph W.,
Koss Leopold G.,
Melamed Myron R.,
Reeder Jay E.,
Weinstein Ronald S.,
Wersto Robert P.,
Wheeless Leon L.
Publication year - 1989
Publication title -
cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.052
H-Index - 304
eISSN - 1097-0142
pISSN - 0008-543X
DOI - 10.1002/1097-0142(19890415)63:8<1592::aid-cncr2820630825>3.0.co;2-g
Subject(s) - flow cytometry , propidium iodide , pathology , coefficient of variation , staining , medicine , microbiology and biotechnology , cytometry , nuclear medicine , chromatography , biomedical engineering , chemistry , biology , biochemistry , apoptosis , programmed cell death
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 transporation 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 cytochemcal 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.