title: Multicolour interphase cytogenetics: 24 chromosome probes, 6 colours, 4 layers.
abstract: From the late 1980s onwards, the use of DNA probes to visualise sequences on individual chromosomes (fluorescent in-situ hybridisation - FISH) revolutionised the study of cytogenetics. Following single colour experiments, more fluorochromes were added, culminating in a 24 colour assay that could distinguish all human chromosomes. Interphase cytogenetics (the detection of chromosome copy number in interphase nuclei) soon followed, however 24 colour experiments are hampered for this application as mixing fluorochromes to produce secondary colours produces images that are not easily distinguishable from overlapping signals. This study reports the development and use of a novel protocol, new fast hybridising FISH probes, and a bespoke image capture system for the assessment of chromosome copy number in interphase nuclei. The multicolour probe sets can be used individually or in sequential hybridisation layers to assess ploidy of all 24 human chromosomes in the same nucleus. Applications of this technique are in the investigation of chromosome copy number and the assessment of nuclear organisation for a range of different cell types including human sperm, cancer cells and preimplantation embryos.
authors: School of Biosciences, University of Kent, Giles Lane, CT2 7NJ, Canterbury, UK.
authors: D
authors: D
authors: Ioannou
authors: E J
authors: EJ
authors: Meershoek
authors: A R
authors: AR
authors: Thornhill
authors: M
authors: M
authors: Ellis
authors: D K
authors: DK
authors: Griffin
mesh_terms: D001755:Blastocyst / Q000166:cytology; D002467:Cell Nucleus / Q000737:chemistry / Q000235:genetics; D002877:Chromosomes, Human / Q000737:chemistry* / Q000235:genetics; D003582:Cytogenetics / Q000295:instrumentation / Q000379:methods*; D004247:DNA / Q000032:analysis / Q000737:chemistry; D015342:DNA Probes / Q000032:analysis* / Q000138:chemical synthesis / Q000378:metabolism; D005307:Fertilization in Vitro; D005456:Fluorescent Dyes; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D007399:Interphase / Q000235:genetics; D008297:Male; D011003:Ploidies; D013094:Spermatozoa / Q000166:cytology; D014407:Tumor Cells, Cultured
chemical_list: D015342:DNA Probes; D005456:Fluorescent Dyes; D004247:DNA