1998 Volume 61 Issue 1 Pages 14-20
We used the poly T probe in rat intestinum tenue tissues to elucidate optimal conditions for the rapid in situ hybridization (ISH) method. Following fixation in either 20% neutral buffered formalin solution or Mildform 20 N, the tissues were immersed in either 10% EDTA or K-CX for 1, 6 or 12 days. All laboratory materials including glassware and reagents were treated with diethyl pyrocarbonate. We found when rapid ISH is applied, that 1) tissue fixed with Mildform 20 N should not be immersed in the decalcifying solution. 2) A 10% EDTA solution should be used as the decalcifying solution, and the shorter the decalcification time, the better. 3) When K-CX solution is used, the specimen should be cut into peices as small as possible and immersed in the solution within one day. These results indicate that the rapid ISH method is very effective when used under optimal conditions for the immersion time in the fixative and decalcifying solutions.
