Procedures for tissue preparation embedding and

sectioning 6.1.1 Fixation

Tissues should be quickly fixed with acidic based solutions such as Bouin's solution (Sigma Chemicals Inc.; cat. no. HT10-1-128) by immersion or if possible from animals, by perfusion with 2% paraformaldehyde in PBS or 4% neutral-buffered formalin and subsequent post-fixation in Bouin's solution. (Bouin's is an acidic fixative and thus will cause DNA nicking. Do not use Bouin's if planning to do in situ detection of DNA fragmentation.)

However, very satisfactory immunostaining can often be achieved using routine archival material that was fixed in 10% buffered formalin and then embedded in paraffin. Thus, if such material is available, it is worth a try.

0 0

Post a comment