Alkaline hydrolysis/methylation-acetylation: a new technique for ultrastructural DNA cytochemistry

Abstract

A new technique for the visualization of DNA-containing structures in electron microscopy is described. Samples of glutaraldehyde-fixed bone marrow from rats were subjected to alkaline hydrolysis to remove RNA and the phosphate of phosphoproteins, followed by a combined blockage of protein carboxyl and amino groups through methylation-acetylation. After uranyl acetate staining of epoxy-embedded ultrathin sections, chromatin from all cell types showed a highly selective and intense electron opacity. Staining methods for DNA were also positive in semithin sections. This simple procedure could be very useful in ultrastructural cytochemistry of DNA and chromatin.