Subbing Slides

Pretreatment

• Several methods to wash the slides:

1. Wash with soap (Alquinox) for an hour, shaking
2. Wash with 0.5% SDS for an hour, shaking
3. Soak O/N in ethanol

• Rinse slides in distilled water 4 times shaking for 15 minutes each time.

Subbing

Gelatin/Chrome Alum

• Heat 300 ml of water to 60°C. Add 1.2g gelatin (Porcine Skin type A, bloom 250 or 300, Sigma G2500). After the gelatin is in solution, cool to 40°C. Add 0.12g chromium potassium sulfate (CrK(SO₄)₂·12H₂O). Stir into solution.
• Dip slides into solution 3 times, waiting 1 min in between. Allow slides to dry. Dip again at the end of the day and dry slides overnight. Keep slides covered during drying so dust does not settle onto them.
• Store the slides at -20°C.

Poly-L-Lysine

• Dip slides into 0.3% poly-L-lysine in water for 30 minutes
• Let slides dry on bench or in 60°C oven (make sure they are covered so nothing sticks to them)