Analysis and interpretation of DNA distributions measured by flow cytometry

Journal Article · Fri Jan 01 04:00:00 EST 1982 · Cytometry (Baltimore); (United States)

DOI:https://doi.org/10.1002/cyto.990030309· OSTI ID:5025863

Dean, P N; Gray, J W; Dolbeare, F A

A principal use of flow cytometers is for the measurement of fluorescence distributions of cells stained with DNA specific dyes. A large amount of effort has been and is being expended currently in the analysis of these distributions for the fractions of cells in the G/sub 1/, S, and G/sub 2/ + M phases of the cell cycle. Several methods of analysis have been proposed and are being used; new methods continue to be introduced. Many, if not most, of these methods differ only in the mathematical function used to represent the phases of the cell cycle and represent attempts to fit exactly distributions with known phase fractions or unusual shapes. In this paper we show that these refinements probably are not necessary because of cell staining and sampling variability. This hypothesis was tested by measuring fluorescence distributions for Chinese hamster ovary and KHT mouse sarcoma cells stained with Hoechst-33258, chromomycin A3, propidium iodide, and acriflavine. Our results show that: a) single measurements can result in phase fraction estimates that are in error by as much as 40% for G/sub 2/ + M phase and 15 to 20% for G/sub 1/ and S phases; b) different dyes can yield phase fraction estimates that differ by as much as 40% due to differences in DNA specificity; c) the shapes of fluorescence distributions and their interpretation are very dependent on the dye being used and on its binding mechanism. 7 figures, 2 tables.

• Research Organization: Lawrence Livermore National Lab., CA
• DOE Contract Number: W-7405-ENG-48
• OSTI ID: 5025863
• Journal Information: Cytometry (Baltimore); (United States), Journal Name: Cytometry (Baltimore); (United States) Vol. 3:3; ISSN CYTOD
• Country of Publication: United States
• Language: English

Related Subjects

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY
400500 -- Photochemistry
550300* -- Cytology
59 BASIC BIOLOGICAL SCIENCES
ACCURACY
ANIMAL CELLS
ANIMALS
BODY
CELL FLOW SYSTEMS
CHEMICAL ANALYSIS
COMPUTER CALCULATIONS
DATA
DATA PROCESSING
DISEASES
DNA
DYES
ELECTROMAGNETIC RADIATION
ENERGY-LEVEL TRANSITIONS
EXCITATION
EXPERIMENTAL DATA
FEMALE GENITALS
FLUORESCENCE
GONADS
HAMSTERS
INFORMATION
LASER RADIATION
LUMINESCENCE
MAMMALS
MICE
MONITORING
NEOPLASMS
NUCLEIC ACIDS
NUMERICAL DATA
ORGANIC COMPOUNDS
ORGANS
OVARIES
PROCESSING
QUANTITATIVE CHEMICAL ANALYSIS
RADIATIONS
RODENTS
SARCOMAS
ULTRAVIOLET RADIATION
VERTEBRATES