用DAPI进行细胞染色

用DAPI进行细胞染色

DAPI的配置

Stock solution of 1mg/ml in ddH2O; working solution of 0.25mg/ml to 50mg/ml in ddH2O or PGM.

1.Place sample on slide.

2.Add a few drops working solution.

3.Squash and view.

Staining � DAPI Staining for Cells:

Separately DAPI staining:

• Wash the cells twice with PBS

• Incubate with 1: 750 of stock DAPI for 5 min in dark

• Wash 2 x 5 min with PBS in dark

• Rinse with water for a few seconds

• Air dry at RT in dark

• Mount with antifade solution (Vectashield, Co#: H-1000, Vector Lab)

5 µl for each d12 mm coverslip, 25 µl for square coverslip and 50µl for full size coverslip

• Seal the coverslip with nail polish

• Store in 4 C up to 2 weeks

DAPI stock

• 4’, 6-Diamidino-2-phenylindole dihydrochloride hydrate (DAPI) from Sigma (Cat# D-9542), 1 mg 1mg/0.5 ml in H2O and stocked at �20 in the dark

Staining with antifade mounting solution:

• Wash the cells twice with PBS

• Mount with antifade solution with DAPI (Vectashield, Co#: H-1200，Vector Lab, 1.5 µg/ml) at 1:3 dilution with Vectashield H-1000 Volume used as above

• Store in 4 C up to 2 weeks