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

B. Trask, G. Van den Engh, R. Nussbaum, C. Schwartz, Joe Gray

Research output: Contribution to journalArticle

31 Citations (Scopus)

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 meausred 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 predicted from cytogenetics and/or probe analyses. Deletions of 3-5 Mb, which were at and below the detection limits of conventional cytogenetics, could be quantified by flow karyotyping in individuals with X-linked diseases such as Duchenne muscular dystrophy, choroideremia, and ocular albinism/ichthyosis. The results show that the use of flow karyotyping to quantify the size of restricted regions of the genome can complement conventional cytogenetics and other physical mapping techniques in the study of genetic disorders.

Original languageEnglish (US)
Pages (from-to)184-195
Number of pages12
JournalCytometry
Volume11
Issue number1
StatePublished - 1990
Externally publishedYes

Fingerprint

Karyotyping
X Chromosome
Aneuploidy
Cytogenetics
Karyotype
Chromosomes
Y Chromosome
DNA
Ocular Albinism
Flow Cytometry
Chromomycins
Choroideremia
Chromomycin A3
Photometry
Ichthyosis
Cell Line
Inborn Genetic Diseases
Duchenne Muscular Dystrophy
Propidium
Base Composition

Keywords

  • aneuploidy
  • chromosome abnormalities
  • deletion
  • Duchenne muscular dystrophy
  • Flow cytometry
  • flow karyotyping
  • human chromosomes
  • ichthyosis
  • ocular albinism
  • steroid sulfatase deficiency

ASJC Scopus subject areas

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

Cite this

Quantification of the DNA content of structurally abnormal X chromosomes and X chromosome aneuploidy using high resolution bivariate flow karyotyping. / Trask, B.; Van den Engh, G.; Nussbaum, R.; Schwartz, C.; Gray, Joe.

In: Cytometry, Vol. 11, No. 1, 1990, p. 184-195.

Research output: Contribution to journalArticle

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