Selective extraction of specific cell components by enzyme or acid hydrolysis is possible from ultrathin sections for electron microscopy and parallel 2 µ sections for light microscopy of tissues fixed in formalin and embedded in a water-soluble polyepoxide, product X133/2097. Normal rat tissues fixed 15 minutes in formalin at 3°C are more rapidly digested by proteinases than those fixed for the same length of time at 20°C. Trypsin selectively attacks the nuclear chromatin and the ribonucleoprotein particles of the ergastroplasm, whereas mitochondria and zymogen granules resist tryptic digestion. Pepsin rapidly attacks the mitochondria and zymogen granules. The ergastoplasm and nucleus at first resist peptic digestion, but in time the entire cytoplasm and interchromatinic portion of the nucleus are attacked. Ribonuclease abolishes cytoplasmic basophilia in 2 µ sections, but parallel ultra-thin sections, stained with uranyl acetate and examined in the electron microscope, show no change in the ribonucleoprotein particles of the ergastoplasm. Desoxyribonuclease alone had no effect, but after pretreatment of the sections with pepsin or hydrochloric acid, desoxyribonuclease specifically attacked the nuclear chromatin. Nucleic acid-containing structures in the sections are gradually disintegrated by perchloric acid or hydrochloric acid.
Article|
July 01 1961
ULTRASTRUCTURAL CYTOCHEMISTRY : Enzyme and Acid Hydrolysis of Nucleic Acids and Protein
Elizabeth H. Leduc,
Elizabeth H. Leduc
From the Institute of Cancer Research, Villejuif, France.
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Wilhelm Bernhard
Wilhelm Bernhard
From the Institute of Cancer Research, Villejuif, France.
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Elizabeth H. Leduc
From the Institute of Cancer Research, Villejuif, France.
Wilhelm Bernhard
From the Institute of Cancer Research, Villejuif, France.
Dr. Leduc's present address is Department of Biology, Brown University, Providence
Received:
January 27 1961
Copyright, 1961, by The Rockefeller Institute Press
1961
J Biophys and Biochem Cytol (1961) 10 (3): 437–455.
Article history
Received:
January 27 1961
Citation
Elizabeth H. Leduc, Wilhelm Bernhard; ULTRASTRUCTURAL CYTOCHEMISTRY : Enzyme and Acid Hydrolysis of Nucleic Acids and Protein . J Biophys and Biochem Cytol 1 July 1961; 10 (3): 437–455. doi: https://doi.org/10.1083/jcb.10.3.437
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