1. Fix tissue in 4% paraformaldehyde in PBS at 4°C overnight (or 4 h at room temperature).
2. Successively replace solution with the following (at least 30 min each with occasional agitation, at room temperature except where stated).
c. 70% Ethanol twice.
f. 100% Ethanol twice.
g. Histolene twice.
h. 1:1 Histolene:melted paraffin wax for 20 min at 60°C.
3. Transfer tissues to a mould at 60°C, orientate and allow wax to set.
4. Cut ribbons of 6 |im sections on a microtome. Float the ribbons of sections on a bath of distilled water at 40°C until free from creases. Collect on subbed slides. Dry slides at 37°C overnight.
Was this article helpful?