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Flow cytometric DNA‐histogram analysis: Non‐stoichiometric fluorochrome binding and pseudo‐aneuploidy
Author(s) -
Kubbies Manfred
Publication year - 1992
Publication title -
the journal of pathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.964
H-Index - 184
eISSN - 1096-9896
pISSN - 0022-3417
DOI - 10.1002/path.1711670411
Subject(s) - aneuploidy , dna , flow cytometry , histogram , flow (mathematics) , stoichiometry , chemistry , biology , microbiology and biotechnology , genetics , physics , computer science , mechanics , chromosome , gene , organic chemistry , artificial intelligence , image (mathematics)
Detection of aneuploid subpopulations using flow cytometry requires stoichiometric binding of nucleic acid‐specific fluorochromes onto DNA. It is shown that parameters like cell type specificity and differentiation stage, cell cycle stage, loss of DNA‐integrity, cell preparation, and cytochemistry affect fluorochrome binding to DNA and give rise to the appearance of pseudo‐aneuploid cell populations. Intercalating as well as non‐intercalating fluorochromes show non‐stoichiometric DNA‐labelling in cell populations with identical DNA content, and pseudo‐aneuplpidy was found in flow cytometers equipped with either arc lamps or argon lasers. Pseudo‐aneuploidy was never observed with intercalating and non‐intercalating fluorochromes within identical specimens, consisting of cells of various differentiation states (e.g., bone marrow) or containing large numbers of dead cells. Therefore, fluorochromes exhibiting different base‐pair specificities or steric binding modes should be applied to be sure of the correct interpretation of small levels of hypo‐ or hyper‐diploidy (±xs 20 per cent).