Measurement and characterization of micronuclei in exfoliated human cells by fluorescence in situ hybridization with a centromeric probe.
Titenko-Holland-N; Moore-LE; Smith-MT
Mutat Res 1994 Feb; 312(1):39-50
A novel method for the determination of micronuclei in human exfoliated cells using fluorescent propidium-iodide (25535164) staining with fluorescence in-situ hybridization (FISH) with a centromeric probe was described. Exfoliated buccal and urothelial cells collected from five healthy unexposed female volunteers were examined for micronuclei using the Feulgen-Fast-Green staining technique as well as the new fluorescent technique. Comparative results were seen using both techniques. The mean percentage of cells with micronuclei was found to be 0.145 for buccal cells and 0.083 for bladder cells. A statistically significant interindividual variability in the frequency of micronuclei was seen. The use of FISH with a centromeric probe allowed the differentiation of micronuclei containing whole chromosomes with centromeres from those containing only chromosome fragments. Buccal and urothelial cells were found to contain 44% and 43% centromere negative micronuclei, respectively. Examination of other cytological abnormalities indicated that fluorescent staining gave more accurate scoring compared with Feulgen-Fast-Green staining. Fewer than 2% of the cells were found to be pyknotic, "broken eggs" or contain fragmented nuclei while karyorrhexis and karyolysis were seen in up to 21%. The authors conclude that the technique described is useful for scoring micronuclei, for the determination of the mechanism of micronucleus formation, and for the identification of abnormal nuclear events in exfoliated human cells.
NIOSH-Publication; NIOSH-Grant; Training; Laboratory-techniques; Analytical-methods; DNA-damage; Chromosome-damage; Cytopathology; Genotoxic-effects;
Author Keywords: Biomarkers; Buccal cells; Urothelial cells; (Humans); Centrometric probe; Micronuclei; Variability
Biomedical & Environ Hlth Scis University of California 322 Warren Hall Berkeley, Calif 94720
University of California Berkeley, Berkeley, California