Beta-Glactasidase Stain
Histology
scientificprotocols authored over 8 years ago
Solutions
- Solution A
- 5mM potassium ferricyanide, crystalline
- 5mM potassium ferrocyanide, trihydrate
- X-gal Stock Solution
- 40mg/mL in DMSO (100mg in 2.5ml DMSO)
- Final X-gal Solution
- Dilute X-gal Stock Solution 1:30 in Solution A
- (warm Solution A to 37°C to prevent precipitation of X-gal)
Staining Protocol
Start with fixed frozen 10μm tissue sections on pap-penned slides
- Allow slides to completely dry for 1 hour
- Fix tissue with 2% formaldehyde + 0.2% glutaraldehyde for 5 min.
- Rinse with 1xPBS
- Incubate slides in x-gal solution in humidity chamber at 37°C overnight
- Rinse with 1xPBS
- Immerse slides in 100% ethanol until all precipitate dissolves (1-3 hours)
- Place slides in:
- a. Eosin Y 25 seconds
- b. 95% Ethanol 1 minute
- c. 100% Ethanol 1 minute
- d. 100% Ethanol 1 minute
- e. 100% Ethanol 1 minute
- f. Xylene 2 minutes
- g. Xylene 10 dips
- Coverslip slides using Cytoseal 60
