Artifacts associated with mithramycin fluorescence in the clinical detection and quantitation of aneuploidy by flow cytometry

Ethanol-fixed cells stored at 4 degrees C exhibit fixation time-dependent hyperchromatism in comparison with freshly fixed cells when stained with mithramycin and examined by flow cytometry. This hyperchromatism has been found to be temperature-dependent, developing fully within 72 hr at room temper...

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Bibliographic Details
Published in:The journal of histochemistry and cytochemistry 1982-04, Vol.30 (4), p.317-322
Main Authors: Cunningham, RE, Skramstad, KS, Newburger, AE, Shackney, SE
Format: Article
Language:English
Subjects:
Aneuploidy
Flow Cytometry - methods
Humans
Lymphocytes - cytology
Lymphoma - pathology
Neoplasms - pathology
Plicamycin
T-Lymphocytes - cytology
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Summary:Ethanol-fixed cells stored at 4 degrees C exhibit fixation time-dependent hyperchromatism in comparison with freshly fixed cells when stained with mithramycin and examined by flow cytometry. This hyperchromatism has been found to be temperature-dependent, developing fully within 72 hr at room temperature, and within 2 hr at 37 degrees C. Cells from normal donors that are stained with mithramycin exhibit spurious aneuploid peaks. These spurious aneuploid peaks can be eliminated by incubating ethanol-fixed cells at 37 degrees C for 2 hr prior to staining; true aneuploidy is not affected by this procedure. In rare instances, cytoplasmic fluorescence can be observed in mithramycin-stained cells. In addition, unexplained hypochromatism and hyperchromatism can be observed in some clinical samples, particularly in human melanoma. The effects of these unexplained staining artifacts can be minimized or eliminated by adopting strict criteria for the clinical detection of aneuploidy by flow cytometry.
ISSN:0022-1554
1551-5044
DOI:10.1177/30.4.6460801