Quantification of the DNA content of structurally abnormal X chromosomes and X chromosome aneuploidy using high resolution bivariate flow karyotyping

Cytometry
B J TraskJ W Gray

Abstract

Quantification of the Hoechst and chromomycin A3 fluorescence intensities of mitotic human chromosomes isolated from karyotypically normal and abnormal cells was performed with a dual beam flow cytometer. The resultant flow karyotypes contain information about the relative DNA content and base composition of chromosomes and their relative frequencies in the mitotic cell sample. The relative copy number of X and Y chromosomes was determined for 38 normal males and females and 6 cell lines with X or Y chromosome aneuploidy. Flow karyotype diagnoses corresponded with conventional cytogenetic results in all cases. We show that chromosome DNA content can be derived from peak position in Hoechst vs. chromomycin flow karyotypes. These values are linearly related to propidium iodide staining intensity as measured with flow cytometry and to the binding of gallocyanin chrome alum to phosphate groups as measured with slide-based scanning photometry. Cell lines with deleted or dicentric X chromosomes ranging in length from 0.53 to 1.95 times normal were analyzed by using flow cytometry. The measured difference in DNA content between a normal X and each of the structurally abnormal chromosomes was linearly correlated to the difference predi...Continue Reading

References

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Citations

Jan 1, 1990·Cytometry·G van den EnghB Trask
May 1, 1995·TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik·J Doležel, S Lucretti
Feb 1, 1993·TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik·S LucrettiJ Fuchs
Jan 1, 1997·TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik·T SchwarzacherN Miller
Mar 1, 1993·Nature Genetics·F GurrieriS W Scherer
Jun 29, 2007·BMC Genomics·Jennifer L FreemanCharles Lee
Nov 27, 2015·The Journal of Molecular Diagnostics : JMD·Yu WuMin Fang
Oct 3, 2002·Nature Reviews. Genetics·Barbara J Trask
Sep 5, 2008·Current Protocols in Cytometry·Z Darzynkiewicz, G Juan

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