Fluorescence in situ hybridization of metaphase chromosomes in suspension

Citation
H. He et al., Fluorescence in situ hybridization of metaphase chromosomes in suspension, INT J RAD B, 77(7), 2001, pp. 787-795
Citations number
16
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Experimental Biology
Journal title
INTERNATIONAL JOURNAL OF RADIATION BIOLOGY
ISSN journal
0955-3002 → ACNP
Volume
77
Issue
7
Year of publication
2001
Pages
787 - 795
Database
ISI
SICI code
0955-3002(200107)77:7<787:FISHOM>2.0.ZU;2-3
Abstract
Purpose: To describe a new method for FISH analysis of metaphase chromosome sin suspension. Materials and methods: Metaphase chromosomes in suspension were isolated fr om a Chinese hamster x human hybrid cell line. 314-2(1) Y, and a human cell line, GM 130B. During suspension hybridization, specific chromosomes were labeled from these two cell lines using either biotin-labeled human genomic DNA, a directly labeled human pancentromere DNA probe or a chromosome #1 l ocus-specific probe. Results The method allows, for the first time, recover of large numbers of isolated individual hybridized chromosomes with good morphology for both hu man x hamster hybrid and human cell lines. The results showed that 46 73% o f the starting number of total chromosomes can be recovered after a FISH in suspension procedure. The well-preserved morphology of hybridized metaphas e chromosomes allowed (1) rapid detection of individual human and hamster c hromosome aberrations, (2) rapid counting of the painted human chromosomes and (3) fast, clear detection of chromosome region-specific probes. This me thod offers a new tool to assay chromosomes and DNA: it offers the possibil ity to develop new techniques for sorting chromosomes based on FISH signals , for early detection and screening of genetic diseases and for bulk measur ement of both balanced or unbalanced chromosomal exchanges and rearrangemen ts. Conclusion: The potential of the method described should facilitate fast, s ensitive population monitoring, and increase sensitivity of the measurement s in chromosome-based biodosimetry.