Counterstaining human chromosomes for flow karyology.
Isolated human metaphase chromosomes stained with the fluorochromes 4'-6-diamidino-2-phenylindole (DAPI) and chromomycin A3(CA3), and counterstained with nonfluorescent netropsin (NTR), have been analyzed by dual-laser flow cytometry. Counterstaining with NTR reduces DAPI fluorescence except at regions on chromosomes 1,9,15,16, and Y, corresponding to C-band heterochromatin. Bivariate flow karyology of human chromosomes treated with this triple-stain combination resolves chromosomes 1,9, and Y distinctly from the remaining chromosomes and resolves variations between chromosome homologues not detected by staining with propidium iodide (PI) or with the double stain combination Hoechst 33258(HO) and CA3.
D002478:Cells, Cultured; D002871:Chromosome Banding; D002877:Chromosomes, Human / Q000648:ultrastructure*; D005347:Fibroblasts / Q000166:cytology; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D007211:Indoles; D007621:Karyotyping; D008297:Male; D008677:Metaphase; D012867:Skin / Q000166:cytology; D013194:Staining and Labeling
D007211:Indoles; C007293:DAPI