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Alkaline hydrolysis/methylation‐acetylation: a new technique for ultrastructural DNA cytochemistry
Author(s) -
Romero J. B.,
Ferrer J. M.,
Tato A.,
Tandler C. J.,
Llorente A. R.,
Castillo P.,
Stockert J. C.
Publication year - 1991
Publication title -
journal of microscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0022-2720
DOI - 10.1111/j.1365-2818.1991.tb03139.x
Subject(s) - cytochemistry , uranyl acetate , chromatin , ultrastructure , staining , dna , glutaraldehyde , methylation , chemistry , alkaline phosphatase , biochemistry , acetylation , electron microscope , microbiology and biotechnology , dna methylation , biology , chromatography , enzyme , gene expression , anatomy , genetics , physics , gene , optics
SUMMARY 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 phospho‐proteins, 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.