Elimination of Material that Obscures Stained Chromosomes in Squashes of Vicia faba Root Tips

Abstract

A procedure for elimination of cytoplasmic debris from Vicia faba root tip cells is: (1) a root tip previously fixed in 3:1 absolute alcohol-acetic acid and stained by the Feulgen method is placed on a slide and squashed in a small drop of water, (2) a cover slip is applied and the cells are flattened with a hand-operated lever device supplying 35 pounds pressure onto a 22 × 22 mm cover glass, (3) the slide is quick-frozen, the cover slip is removed, and the slide is dropped immediately into water, (4) the slide is cleared through an alcohol-xylene dehydration series and permanently mounted. The significant result of this procedure is the consistent presence of clear, flat cells showing excellent definition of chromosomes.