In-vitro decondensation of human spermatozoa for fluorescence in-situ hybridization.
In order to enhance the efficiency of fluorescence in-situ hybridization (FISH) on human interphase spermatozoa, a simple method for partial decondensation of human spermatozoa chromatin is described. The spermatozoa were washed once in 0.01 M Tris and decondensed using 10 mM dithiothreitol in 0.05 M Tris for 10-50 min, before being spread onto clean slides. Sperm samples were observed every 10 min under a phase-contrast microscope in order to modify the duration of the decondensation process. Using several centromeric or YAC probes in multicolour FISH, high hybridization rates were obtained.
Rousseaux
S
S
Reproductive Biology Unit, DyOGen, Albert Bonniot Institute, Grenoble University Medical School, La Tronche, France.
Chevret
E
E
D000328:Adult; D002843:Chromatin / Q000187:drug effects*; D018244:Chromosomes, Artificial, Yeast; D015342:DNA Probes; D005069:Evaluation Studies as Topic; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D066298:In Vitro Techniques; D008297:Male; D013094:Spermatozoa / Q000187:drug effects*
D002843:Chromatin; D015342:DNA Probes