Computerized measurement of the DNA content, areas, and autoradiographic grains of the same nuclei: demonstration that lightly (/sup 3/H)thymidine-labeled bone marrow cells are predominantly in G0/G1 and G2

Journal Article · Wed Oct 31 23:00:00 EST 1984 · J. Histochem. Cytochem.; (United States)

DOI:https://doi.org/10.1177/32.11.6386975· OSTI ID:5709854

Allison, D C; Meyne, J; Ridolpho, P F; Bose, K; Chakerian, M; Robertson, J

A computerized flying spot microdensitometer and scanning stage have been combined to measure the cellular DNA content, nuclear areas, and autoradiographic grain areas of the same cells. The slide positions of the Feulgen-stained, (/sup 3/H)thymidine-labeled cells are mapped with the computerized stage, and nuclear DNA content and areas are then determined by integral absorbance measurements at 588 nm. Following autoradiograph preparation, the cells are relocated and the areas of the autoradiographic grains over each nucleus are measured at a light wavelength (625 nm) and an optical density setting (greater than 0.10) that do not detect the Feulgen stain. The microcomputer calculates the portion of each nucleus covered with autoradiographic grains (grain area proportion, GAP), and it links the GAP value to the DNA content of each nucleus in the computer file for subsequent sorting and analysis. By using this system in a study of mouse bone marrow cells labeled in vivo with (/sup 3/H)thymidine, it was found that all S-phase cells were clearly labeled after 8 or more days of autoradiographic exposure. Prolonged exposures (up to 64 days) led to detection of lightly labeled cells (0.1 less than GAP less than 0.8) with G1/G0 and G2 DNA content.

• Research Organization: Albuquerque Veterans Administration Medical Center, NM
• OSTI ID: 5709854
• Journal Information: J. Histochem. Cytochem.; (United States), Journal Name: J. Histochem. Cytochem.; (United States) Vol. 32:11; ISSN JHCYA
• Country of Publication: United States
• Language: English

Related Subjects

550301* -- Cytology-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ANIMAL CELLS
ANIMALS
AUTORADIOGRAPHY
AZINES
BIOCHEMISTRY
BONE MARROW CELLS
CELL CONSTITUENTS
CELL CYCLE
CELL NUCLEI
CHEMISTRY
COMPUTER CALCULATIONS
CONNECTIVE TISSUE CELLS
CYTOCHEMISTRY
CYTOLOGICAL TECHNIQUES
DENSITOMETERS
DNA
HETEROCYCLIC COMPOUNDS
IN VIVO
LABELLED COMPOUNDS
LABELLING
MAMMALS
MEASURING INSTRUMENTS
MICE
NUCLEIC ACIDS
NUCLEOSIDES
NUCLEOTIDES
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PHOTOMETERS
PYRIMIDINES
RIBOSIDES
RODENTS
SOMATIC CELLS
THYMIDINE
TRITIUM COMPOUNDS
VERTEBRATES