Mapping nonisotopically labeled DNA probes to human chromosome bands by confocal microscopy

Donna Albertson, Paul Sherrington, Mark Vaudin

Research output: Contribution to journalArticle

Abstract

A method for mapping nonisotopically labeled probes to human metaphase chromosomes that can be used with laser scanning confocal microscopy has been developed. Only a limited number of wavelengths are available from the argon ion lasers used in most commercial instruments and therefore a method that allowed the visualization of bands on human chromosomes stained with propidium iodide and, simultaneously, the detection of hybridization signals using FITC-labeled antibodies was developed. The confocal microscope was used to map single-copy probes to chromosome bands and the positions of the probes on the R-banded chromosomes corresponded to map positions previously determined on Hoechst 33258-stained chromosomes (G-banded). A comparison of confocal imaging of single-copy hybridization signals with conventional fluorescence microscopy and high-sensitivity video cameras revealed little difference in sensitivity but greater resolution of chromosome bands with the confocal microscope. The polymerase chain reaction was used to prepare nonisotopically labeled probes for in situ hybridization and to amplify Alu and KpnI family repeats from cloned DNA to be used to suppress hybridization of these repeat sequences so that a cosmid probe could be mapped to a chromosome band.

Original languageEnglish (US)
Pages (from-to)143-150
Number of pages8
JournalGenomics
Volume10
Issue number1
DOIs
StatePublished - 1991

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DNA Probes
Human Chromosomes
Confocal Microscopy
Chromosomes
Chromosomes, Human, 21-22 and Y
Alu Elements
Bisbenzimidazole
Cosmids
Gas Lasers
Propidium
Fluorescein-5-isothiocyanate
Metaphase
Fluorescence Microscopy
In Situ Hybridization
Polymerase Chain Reaction
Antibodies
DNA

ASJC Scopus subject areas

  • Genetics

Cite this

Mapping nonisotopically labeled DNA probes to human chromosome bands by confocal microscopy. / Albertson, Donna; Sherrington, Paul; Vaudin, Mark.

In: Genomics, Vol. 10, No. 1, 1991, p. 143-150.

Research output: Contribution to journalArticle

Albertson, Donna ; Sherrington, Paul ; Vaudin, Mark. / Mapping nonisotopically labeled DNA probes to human chromosome bands by confocal microscopy. In: Genomics. 1991 ; Vol. 10, No. 1. pp. 143-150.
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