A method for staining 3T3 cell nuclei with propidium iodide in hypotonic solution

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Abstract
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°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°C. Compared to this technique, previously described propidium iodide‐staining methods gave poor resolution with 3T3 cells.