Hoechst 33342 nuclei staining
Last updated: 1/12/12 By: Tim Starr
Stain nuclei with a DNA binding dye that changes fluorescence, and score for pyknotic looking nuclei.
- Hoechst 33342 (Invitrogen Cat# H3570) at 10 mg/ml in H2O
- Cells
- Start with plated cells that have not reached confluence.
- Prepare enough media with Hoechst dye (stock = 10 mg/ml) at the desired concentration
- For 6 well plate, add 6 µl 10mg/ml Hoechst dye to 12 ml media (final 5 µg/ml). Note: Can use from 0.2 to 5 µg/ml Hoechst dye concentration
- Aspirate media from cells
- Wash with PBS
- Add 2 ml media/Hoechst to each well, incubate 20 to 30 minutes at 37C 5% CO2
- Count pyknotic cells and normal cells