Search Thermo Fisher Scientific
Search Thermo Fisher Scientific
Hoechst 33342 nucleic acid stain is a popular cell-permeant nuclear counterstain that emits blue fluorescence when bound to dsDNA. It is also available as a solution.
1. Prepare the Hoechst dye stock solution by dissolving the contents of one vial (100 mg) in 10 mL of deionized water (diH2O) to create a 10 mg/mL (16.23 mM) solution. Note: Hoechst dye has poor solubility in water, so sonicate as necessary to dissolve. The 10 mg/mL Hoechst stock solution may be stored at 2–6°C for up to 6 months or at ≤–20°C for longer periods. |
1. Culture cells in an appropriate medium and vessel for HCA. |
2. Optional: Add a test compound or drug to the cells, and incubate as desired. |
3. Prepare the Hoechst staining solution by adding 5 μL Hoechst stock solution to 10 mL PBS. |
4. Mix well. |
5. Remove the medium. |
6. Add 100 μL staining solution to each well. |
7. Incubate for 5–15 minutes, protected from light. |
8. Optional: You may image directly in the staining solution, if you wish. |
9. Remove the staining solution. |
10. Wash the cells 2–3 times in PBS. |
11. Image the cells. |
Hoechst 33342 | |
---|---|
Excitation/Emission (nm) | 350/461 |
Standard filter set | DAPI |
EVOS Light Cube | DAPI |
Storage conditions | 2–6°C or ≤–20°C |
Cells stained with Hoechst 33342 and imaged with the Thermo Scientific CellInsight High-Content System.