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用DAPI进行细胞染色

发布时间: 2017-09-18  点击次数: 739次

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:


Separay 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