Mapping of single-copy DNA sequences on human chromosomes by in situ hybridization with biotinylated probes: Enhancement of detection sensitivity by intensified-fluorescence digital-imaging microscopy

Journal Article · Sun Jan 01 00:00:00 EST 1989 · Proceedings of the National Academy of Sciences of the United States of America; (USA)

DOI:https://doi.org/10.1073/pnas.86.2.582· OSTI ID:5032042

Viegas-Pequignot, E; Dutrillaux, B; Magdelenat, H ^[1]; Coppey-Moisan, M ^[2]

---

1. Institut Curie, Paris (France)
2. Institut Curie, Paris (France) Institut National de la Sante et de la Recherche Medicale, Orsay (France)

Two single-copy DNA segments of 6 kilobases (kb) and 2.3 kb were labeled with biotin-labeled dUTP (Bio-11-dUTP) and hybridized to human chromosomes. These probes were detected by immunofluorescence and directly mapped on chromosomes by using classical fluorescence microscopy and a microchannel-plate-intensified video camera. By a subsequent R-banding, the 6-kb and 2.3-kb fragments were precisely localized to the 18p11.3 band and to the 22q11.2 band, respectively, in agreement with previous results obtained with radioactive probes. The adaptation of fluorescence intensification and digital image processing (frame integration to enhance signal-to-noise ratio and linear contrast stretching) to microscopy makes it possible to detect very weak fluorescent spots on chromosomes. This system allows a high spatial resolution, even at very low fluorescence levels. The efficiency and the specificity of the hybridization and detection methodology give a direct and precise localization of the short single-copy sequences on human chromosomes.

• OSTI ID: 5032042
• Journal Information: Proceedings of the National Academy of Sciences of the United States of America; (USA), Vol. 86:2; ISSN 0027-8424
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
CHROMOSOMES
GENETIC MAPPING
DNA
MICROSCOPY
BIOTIN
DNA HYBRIDIZATION
DNA SEQUENCING
FLUORESCENCE
IMAGE PROCESSING
AZOLES
CARBOXYLIC ACIDS
HETEROCYCLIC ACIDS
HETEROCYCLIC COMPOUNDS
HYBRIDIZATION
IMIDAZOLES
LUMINESCENCE
MAPPING
NUCLEIC ACIDS
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
ORGANIC SULFUR COMPOUNDS
PROCESSING
STRUCTURAL CHEMICAL ANALYSIS
VITAMIN B GROUP
VITAMINS
550200* - Biochemistry