Analysis of meiotic segregation in human nondecondensed interphase spermatozoa by triple colour rapid direct fluorescent in situ hybridization.
Meiotic segregation of chromosomes X,Y and 1 was analyzed by triple colour rapid fluorescent in situ hybridization (FISH) with directly labelled probes on 4506 non-decondensed and non-cleaned interphase spermatozoa from four healthy male donors to test the possibility of rapid sperm FISH by omitting the conventional sperm decondensation and cleaning steps. Only 0.15 per cent of sperms were without any signals which suggested high hybridization success. An abnormal number of signals was seen in 1.6 per cent of sperms. Chromosome specific as well as donor specific segregation error was seen similar to previous reports. There was a wide variation in the ratio of normal X and Y bearing sperm from donor to donor. This study indicated that for segregation studies sperm FISH can be carried out in three hours with directly labelled probes without the steps for separation of sperm from somatic cells (cleaning), sperm swelling and sperm DNA decondensation.
Halder
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Galton Laboratory, Department of Human Genetics, UCL, London, UK.
Tutscheck
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B
D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D007399:Interphase; D008297:Male; D008540:Meiosis; D012680:Sensitivity and Specificity; D013094:Spermatozoa / Q000648:ultrastructure*