A method for staining 3T3 cell nuclei with propidium iodide in hypotonic solution.
A modification of the propidium-iodide hypotonic sodium citrate method has been developed specifically for high-resolution staining of mouse 3T3 cell nuclei for analysis by flow cytometry. The method employs a brief treatment of cells at 37 degrees C with Triton X-100 and RNAse in the presence of propidium iodide in hypotonic sodium citrate, followed by restoration to isotonicity with NaCl. The average CV obtained for the G1 peak was 3.5%, and the samples were stable for 1-2 weeks at 4 degrees C. Compared to this technique, previously described propidium iodide-staining methods gave poor resolution with 3T3 cells.
D000818:Animals; D002467:Cell Nucleus / Q000032:analysis*; D002478:Cells, Cultured; D004247:DNA / Q000032:analysis*; D005453:Fluorescence; D007038:Hypotonic Solutions; D051379:Mice; D008807:Mice, Inbred BALB C; D009994:Osmolar Concentration; D010617:Phenanthridines; D011419:Propidium; D012260:Ribonucleases / Q000494:pharmacology; D013194:Staining and Labeling / Q000379:methods*
D007038:Hypotonic Solutions; D010617:Phenanthridines; D011419:Propidium; D004247:DNA; D012260:Ribonucleases