A fluorescence in situ hybridization technique for retrospective cytogenetic analysis

Abstract

Fluorescence in situ hybridization (FISH) is being used increasingly in clinical practice; however, current FISH techniques require fresh material, and there is considerable variation in hybridization efficiency between laboratories. We have modified a FISH technique described by Pinkel et al. (1986) that works not only on freshly G-banded material but also on cytogenetic preparations ranging in age from 2 wk to 12yr. We have tested this technique on several centromeric alphoid satellite probes (D1Z5, D7Z1, D17Z1, DXZ1, and DYZ3) and one noncentromeric minisatellite probe (D1Z2). Our average hybridization efficiency on freshly banded preparations for these probes is consistently greater than 90 %. The combination of higher efficiency and the ability to perform hybridization on previously G-banded material makes this a valuable technique for retrospective analyses.

Clarification of subtle reciprocal rearrangements using fluorescence in situ hybridization

“…Centromere-specificDNA probes are most useful in interphase analysis of aneuploidy ; however, elucidation of structural abnormalities, such as translocations, can be better achieved by applying chromosome-specific DNA libraries [Pinkel et al, 1988;Tkachuk et al, 1991;Smit et al, 1991;Babu and Wiktor, 1991;Kuo et al, 19911. Such libraries are also useful for detecting aneuploidy and interpreting complex karyotypes in tumors [Cremer et al, 1988;Kibbelaar et al, 1991;Smit et al, 1990, 19911.…”

Induction of senescence and control of tumorigenicity in BK virus transformed mouse cells by human chromosome 6

“…Sequential GTG banding and genomic painting were performed according to Smit et al (1990), Babu and Wiktor (1991), and Klever et al (1991) with minor modifications. After trypsin treatment and staining with 5% Giemsa, GTG banded preparations were embedded in Eukitt and photographed.…”

Fluorescent in situ hybridization on archival G-banded slides

“…The procedure works well for fresh GTG-banded preparations with out mounting prior to CISS hybridization. Babu and Wiktor (1991) described a fluorescent in situ hybridization technique for use with archival slides which involved repeat-sequence and single-copy probes. Their article did not report whether this procedure also works for whole chromosome painting probes.…”