A Technique for combining the Staining of Fixed Nuclear Material with the Subsequent Separation of Plant Tissue into Individual Cells

Abstract

FOLLOWING an earlier investigation¹, a further study has been made of the mode of action of iso-propyl-phenylcarbamate. In order that the observed inhibitory effects on the elongation of Zea mays roots could be interpreted in terms of cell division and cell extension, it was necessary to evolve a technique which coupled staining of the nuclear material with the separation of root tips into the individual component cells. The method, developed by Brown² for Pisum sativum, of gently bombarding carminestained root tips with glass beads, failed with maize roots to give adequate cell separation unless the period of bombardment was greatly prolonged, and then too many of the cells were damaged; also the differential staining of the nucleus and cytoplasm was not good enough to allow of the detection of the earliest stages of prophase. The alternative of using a pectinase³ was then investigated, and eventually the following procedure for the separation of previously stained tissue was found to give excellent results if ‘Pectinase’ (Nutritional Biochemicals Company, Ohio) was employed.