Troubleshooting Sections That Crumble and a Specimen That Shatters When Cutting

Sometimes a specimen being cut on a microtome may shatter or have sections that crumble. The problem can be resolved if the cause is identified.

Situation: sections crumble, specimen tears out, specimen shatters

Differential diagnosis:

(1) The specimen did not undergo adequate dehydration.

(2) The clearing step was not done properly

(3) There were problems with the paraffin bath.

(4) Paraffin is not the best choice as the embedding material.

Cause	Fiinding	Solution
incomplete dehydration	tissue soft and mushy	reprocess the specimen
alcohol not completely removed by the clearing fluid		make sure that the alcohol is cleared before embedding
choice of clearing fluid	specimen hard and brittle when xylene used	try toluene or a mixture of toluene plus cedar oil
too much time spent in paraffin bath		limit time spent in the paraffin bath
paraffin bath too hot		lower the temperature
specimen too hard for paraffin	specimen shatters and falls out of the paraffin	embed in a harder wax
specimen fragile	specimen shatters	embed in celloidin or other material

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