Crystalline violet staining

Summary

This lab was derived from: Animal Cell Culture - A Guide to Basic Techniques (5th Edition)

Operation method

Scheme 16.3 Crystalline Violet Staining

Materials and Instruments

D-PBSA D-PBSA:Methanol (1:1)
Methanol Crystalline violet
Filtering funnel, filter paper, bottle for recovering the dye solution.

Move

1. Aspirate and discard the medium.

2. Rinse the monolayer with D-PBSA and discard the rinse solution.

3. Add 5 ml of D-PBSA/Methanol per 25 ^cm2, let stand for 2 min, discard D-PBSA/Methanol.

4. Add 5 ml of fresh methanol and leave for 10 min.

5. Discard the methanol, empty the petri dish and let it dry.

6. Add 5 ml of crystal violet per 25 ^cm2 and let stand for 10 min.

7. Place a filtering funnel (sized to hold 5 ml of dye solution per petri dish) on the bottle.

8. Pour the solution into the funnel.

9. Rinse the petri dishes with tap water, then with deionized water and dry the petri dishes.

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