Crystal Violet staining stains nuclei a deep purple color, aiding in their visualization. It can also be used to visualize colonies of cells. The entire staining protocol takes less than an hour.
- Place cells on ice and wash 2X with cold PBS (keep in refrigerator).
- Fix for 10 minutes with ice-cold 100% methanol (keep in freezer).
- Aspirate methanol from plates. If you want to stop here, cover the cells with 50% glycerol in PBS, wrap the plate with parafilm or plastic wrap, and keep in the refrigerator. They will be find for at least a few weeks.
- Move the cells off ice to room temperature and add cover them with 0.5% crystal violet solution in 25% methanol. Incubate for 10 minutes. Remove the crystal violet---it can be reused or disposed of as toxic waste.
- Wash the cells in water several times, until the dye stops coming off. A quick way to do this is to submerge the entire dish in a container full of water, then dump the water out and refill it a few times.
- Allow the cells to dry at room temperature (maybe overnight). They can be stored for a long time at room temperature.
- Always pipette on the sides of the dish to avoid detaching cells
- Crystal violet is toxic and should be handled carefully.
- 500 mg Crystal Violet
- Store at room temperature
