Multiple colors by fluorescence in situ hybridization using ratio-labelled DNA probes create a molecular karyotype

Abstract

Fluorescence in situ hybridization (FISH) is now widely used for the localization of genomic DNA fragments, and the identification of chromosomes by painting. We now show that half of the chromosomal complement can be painted in twelve different colors by using human chromosome specific libraries carrying three distinct labels mixed in multiple ratios. The photographs are in 'real' color rather than 'colorized'. The painting technique described here can be used for the identification of small or complex chromosomal rearrangements and marker chromosomes in humans or in any other species for which well defined chromosome specific libraries exist in a laboratory equipped with a conventional fluorescence microscope. The versatility of this novel cytogenetic technology may well constitute an advancement comparable to the introduction of chromosome banding and high resolution analysis of chromosomes in prometaphase.

Chromosomal bar codes produced by multicolor fluorescence in situ hybridization with multiple YAC clones and whole chromosome painting probes

“…As a result, it can be anticipated that multiple color FISH should become a routine procedure in cytogenetic laboratories. While microphotographs showing 'real color pictures' (25) can be taken directly on color slides as demonstrated by Dauwersee et al (20), approaches for the quantitative evaluation of multicolor FISH by digital fluorescence microscopy and image analysis have also been developed (22,26) and will become indispensable for the automated evaluation of CBCs (see below). A number of Alu-PCR amplified YACs used for the present experiments yielded chromosomal bars on target chromosome bands of both homologs and both chromatids in all evaluated metaphase spreads.…”

“…These authors realized the simultaneous visualization of four chromosome-specific repetitive DNA probes with different colors. More recently, Ried et al have applied combinatorial FISH and digital fluorescence microscopy for the multicolor painting of six chromosomes (19), while Dauwerse et al achieved the simultaneous visualisation of twelve different chromosome pairs using chromosome specific DNA-libraries labeled with different ratios of three haptens (20).…”

Human Synaptonemal Complex Protein 1 (SCP1): Isolation and Characterization of the cDNA and Chromosomal Localization of the Gene

“…The metaphases were accumulated in EBV transformed human lymphocytes by the thymidine synchronization frame and is preceded by a consensus sequence for eumethod (Viegas-Péquignot, 1993) and spread onto slides as described karyotic translation initiation (Kozak, 1986). Furtherpreviously (Dauwerse et al, 1992). In situ hybridization was permore, in the amino acid sequence alignment of hsSCP1 formed as described by Dauwerse et al (1992).…”

“…Furtherpreviously (Dauwerse et al, 1992). In situ hybridization was permore, in the amino acid sequence alignment of hsSCP1 formed as described by Dauwerse et al (1992). Briefly, the procedure and three rodent SCP1 proteins (Fig.…”

A strategy for the characterization of minute chromosome rearrangements using multiple color fluorescence in situ hybridization with chromosome-specific DNA libraries and YAC clones

“…Here, we describe a strategy to define the origin of genetic material in marker chromosomes even in cases where this origin cannot be resolved by using conventional chromosome analyses. This strategy is based on recent advances in multicolor chromosome painting (Nederlof et al 1990(Nederlof et al , 1992Ried et al 1992a, b;Dauwerse et al 1992;Lengauer et al 1993).…”