Preparation and bivariate analysis of suspensions of human chromosomes

Journal Article · Tue Jan 01 00:00:00 EST 1985 · Cytometry (Baltimore); (United States)

DOI:https://doi.org/10.1002/cyto.990060203· OSTI ID:5887117

van den Engh, G J; Trask, B J; Gray, J W; Langlois, R G; Yu, L C

Chromosomes were isolated from a variety of human cell types using a HEPES-buffered hypotonic solution (pH 8.0) containing KCl, MgSO/sub 4/ dithioerythritol, and RNase. The chromosomes isolated by this procedure could be stained with a variety of fluorescent stains including propidium iodide, chromomycin A3, and Hoeschst 33258. Addition of sodium citrate to the stained chromosomes was found to improve the total fluorescence resolution. High-quality bivariate Hoeschst vs. chromomycin fluorescence distributions were obtained for chromosomes isolated from a human fibroblast cell strain, a human colon carcinoma cell line, and human peripheral blood lymphocyte cultures. Good flow karyotypes were also obtained from primary amniotic cell cultures. The Hoeschst vs. chromomycin flow karyotypes of a given cell line, made at different times and at dye concentrations varying over fourfold ranges, show little variation in the relative peak positions of the chromosomes. The size of the DNA in chromosomes isolated using this procedure ranges from 20 to 50 kilobases. The described isolation procedure is simple, it yields high-quality flow karyotypes, and it can be used to prepare chromosomes from clinical samples. 22 references, 7 figures, 1 table.

• Research Organization: Erasmus Univ., Rotterdam, Netherlands
• DOE Contract Number: W-7405-ENG-48
• OSTI ID: 5887117
• Journal Information: Cytometry (Baltimore); (United States), Vol. 6
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
CHROMOSOMES
MULTIVARIATE ANALYSIS
SAMPLE PREPARATION
DNA
EXPERIMENTAL DATA
FIBROBLASTS
FLUORESCENCE
KARYOTYPE
LYMPHOCYTES
STAINS
TUMOR CELLS
ANIMAL CELLS
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY FLUIDS
CONNECTIVE TISSUE CELLS
DATA
INFORMATION
LEUKOCYTES
LUMINESCENCE
MATERIALS
MATHEMATICS
NUCLEIC ACIDS
NUMERICAL DATA
ORGANIC COMPOUNDS
SOMATIC CELLS
STATISTICS
550300* - Cytology