Beta-Glactasidase Stain

### Solutions 1. 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 1. 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