Alkaline hydrolysis/methylation-acetylation: a new technique for ultrastructural DNA cytochemistry.
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.
Romero
J B
JB
Departamento de Biología Animal y Biología Celular, Facultad de Biología, Universidad de Valencia, Spain.
Ferrer
J M
JM
Tato
A
A
Tandler
C J
CJ
Llorente
A R
AR
Del Castillo
P
P
Stockert
J C
JC
D000107:Acetylation; D000818:Animals; D001853:Bone Marrow / Q000378:metabolism / Q000648:ultrastructure*; D002843:Chromatin / Q000648:ultrastructure*; D004247:DNA / Q000648:ultrastructure*; D006868:Hydrolysis; D008745:Methylation; D008854:Microscopy, Electron / Q000379:methods*; D051381:Rats; D011919:Rats, Inbred Strains
D002843:Chromatin; D004247:DNA