Isopropyl-Alcohol-Paraffin Methods for Plant Tissues

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Abstract
Two methods using isopropyl alcohol for dehydration prior to paraffin infiltration of plant tissues are reported: (1) a modified Rawlins-Takahashi (1947) schedule in which the preliminary dehydration is effected by concentrating glycerol and (2) isopropyl alcohol as the sole agent for dehydration. With the latter method the fixed tissues are dehydrated successfully in 60%, 85%, and 99% isopropyl alcohol. Paraffin infiltration is accomplished by placing the tissues in isopropyl alcohol over solid paraffin in a vial and heating to 56[degree]-58[degree]C. The tissues settle into the melted paraffin as infiltration progresses. Several changes of pure paraffin are then made, with the last change under reduced pressure. The embedded tissues are trimmed and soaked 2-4 hours at 40[degree]C in either water or a glycerol, acetic acid, 70% alcohol mixture (10:15:75) to reduce static and insure uniform ribboning during microtomy.