A New and Rapid Method for Making Permanent Aceto-Carmin Smears

Abstract

Smear the pollen mother cells of a single anther from each bud on a clean dry slide, using a small scalpel. Flood the slide with Belling''s acetocarmin and heat for a second over an alcohol flame. Examine under the microscope to determine stage of microsporogenesis. Smear the remaining anthers in the same manner, but fix and stain them by immediate immersion, face downward, in a Petri dish full of hot (steaming) acetocarmin for 1-10 min. Rapldly transfer through the following mixtures: 2 parts 99% (glacial) acetic acid plus 1 part absolute ethyl alcohol, 1 part acetic acid plus 2 parts absolute alcohol, and, finally, 1 part acetic acid plus 9 parts absolute alcohol. Dehydrate completely by 1-2 minutes'' immersion in pure absolute alcohol, and clear 2-3 minutes in equal parts of xylene and absolute alcohol. Mount in balsam. The whole process takes 5-15 min. and is particularly recommended for chromosome counts.