Prestaining of membrane markers to identify specific areas for Feulgen cytophotometric determinations in a single section

In cytospectrophotometric determinations of the nuclear DNA content in tissues, two consecutive sections are commonly employed: one stoichiometrically stained (as with the Feulgen reaction) for the actual measurements and a second routinely stained (as with hematoxylin and eosin) to define the limits of abnormal areas. This paper proposes the use of stainable cell membrane markers to identify the boundaries of such areas in only the one section in which DNA measurements are to be performed. The use of this procedure for the analysis of enzyme-altered foci and preneoplastic nodules in the rat liver is described. The membrane marker staining, which does not affect the nucleus or cytoplasm, does not interfere with the nuclear DNA determinations.