Measurement of mammalian sperm deoxyribonucleic acid by flow cytometry. Problems and approaches

Journal Article · Sat Jan 01 04:00:00 EST 1977 · J. Histochem. Cytochem.; (United States)

DOI:https://doi.org/10.1177/25.7.70455· OSTI ID:6543080

Van Dilla, M A; Gledhill, B L; Lake, S; Dean, P N; Gray, J W; Kachel, V; Barlogie, B; Goehde, W

Measurement of mammalian sperm deoxyribonucleic acid content is of importance in several areas of biomedical research. When measured in flow systems with orthogonal axes of illumination, flow and detection, an unexpected, distorted distribution consisting of a narrow peak with a lateral extension to the right is observed. Several lines of evidence lead to the conclusion that this effect is an optical-geometric artifact attributable to the flat shape and high index of refraction of mammalian sperm heads. This artifact disappears when an epi-illumination flow system is used in which the optic axes for illumination and detection and the flow axis are all coincident. Other approaches also eliminate the artifact. The resulting coefficients of variation observed after acriflavine-Feulgen staining are 4-5%, short of the goal of 1.5% required to distinguish between human sperm bearing X and Y chromosomes and to develop a mutagen test system using mice.

• Research Organization: Univ. of California, Livermore
• OSTI ID: 6543080
• Journal Information: J. Histochem. Cytochem.; (United States), Journal Name: J. Histochem. Cytochem.; (United States) Vol. 25:7; ISSN JHCYA
• Country of Publication: United States
• Language: English

Related Subjects

550300 -- Cytology
550400* -- Genetics
550600 -- Medicine
59 BASIC BIOLOGICAL SCIENCES
62 RADIOLOGY AND NUCLEAR MEDICINE
ANIMALS
BIOMEDICAL RADIOGRAPHY
CELL FLOW SYSTEMS
DIAGNOSIS
DIAGNOSTIC TECHNIQUES
DNA
FLUOROSCOPY
GAMETES
GERM CELLS
MAMMALS
MEDICINE
MICE
MUTAGENESIS
MUTATIONS
NUCLEAR MEDICINE
NUCLEIC ACIDS
ORGANIC COMPOUNDS
RADIOLOGY
RODENTS
SPERMATOZOA
VERTEBRATES