De-paraffinizing for staining

Materials:

Xylene

Ethanol series (100%, 95%, 85%, 70%, 50%, 30%)

PBS

4% Formaldehyde

0.1M Triethanolamine

Acetic anhydride

Procedure:

1) Dewax slides in xylene for 10 min

2) Remove xylene with 100% ethanol for 2 min

3) Rehydrate by putting slides quickly through 100%, 95%, 85%, 70%, 50%, then 30% ethanol

4) Remove ethanol

5) Rinse in PBS for 5 min

6) Post-fix sample in 4% formaldehyde diluted in PBS for 20 min

7) Wash in PBS twice for 5 min

8) To 250 mL 0.1M triethanolamine add 0.625 mL acetic anhydride, mix well on stirrer

9) Dip slides in water and then immerse in acetic anhydride solution for 10 min

Wash with PBS twice for 5 min