The alkaline hydrolysis of nucleic acid for removal of RNA associated fluorescence in phenantridium related flow through cytofluorometry

Abstract

Flow through microfluorometry of phenantridium and fluoresceinisothiocyanate stained HeLa cells showed the absence of extranuclear RNA related fluorescence after prolonged hydrolysis of cells in ethanolic barium hydroxide. DNA distribution were as good as in RNAse treated samples. Distribution pattern of cellular protein/DNA ratios yielded greater resolution power than those after RNAse digestion. The cellular protein content has not been affected. The ethanolic hydrolysis has the advantage to be of less expense and to allow cell conservations for further measurements with other well preserving fixatives. Thus, application of glutaraldehyde has been proved to be valuable.