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PMID: 2643118 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

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.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 86 ·No. 2 ·1989-01-00 ·Pages 582-6

Viegas-Pequignot E, Dutrillaux B, Magdelenat H, Coppey-Moisan M

Abstract

Two single-copy DNA segments of 6 kilobases (kb) and 2.3 kb were labeled with biotin-labeled dUTP (Bio11-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 (less than 0.6 micron), 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.

MeSH Terms

Base Sequence Biotin Chromosome Mapping Chromosomes, Human DNA/genetics DNA Probes Fluorescent Antibody Technique Humans Image Processing, Computer-Assisted Microscopy, Fluorescence Nucleic Acid Hybridization

Chemicals

DNA Probes Biotin DNA

Authors & Affiliations

4 authors, click to expand affiliations / ORCID

Viegas-Pequignot E

U.R. 620, Section de Biologie, Institut Curie, Paris, France.

Dutrillaux B

Magdelenat H

Coppey-Moisan M

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19 references, click to expand

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Article Info

Journal

Proceedings of the National Academy of Sciences of the United States of America

Abbr.

Proc Natl Acad Sci U S A

ISSN

0027-8424

DOI

10.1073/pnas.86.2.582

Published

1989-01-00

Pages

582-6

Language

English

Region

United States

NLM ID

7505876

PMCID

PMC286516

Subset

External Links

PubMed source DOI: 10.1073/pnas.86.2.582 PMC Full Text View journal details

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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.

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