On the New Approach of Tannic Acid and Digitonine to the Biological Fixatives

Abstract
Radioactive labelled cholesterol is fixed in the tissue as well as labelled other 3β-OH group using saturated digitonine (Digt) in each of the fixative, washing solutions, dehydrating alcohole and propylenoxide. Free metabolites from the 3β-OH group which have already not had 3β-OH structure, can be any more fixed in the tissue. Some of such metabolites may be combined with proteins including lipo- or glyco-proteins. But low molecular proteins should not be fixed with common fixatives, such as aldehydes and OsO4. From such stand-point, we have examined to approach the tannic acid(Tann) for the purpose of protein-fixation, as follows: l) 0.5, 1, 2, 4, 8% Tann and 2.5% glutaraldehyde(Glut) in 0.05M phosphate buffer at pH 6.8, 0°C, for 1.5 to 2 hrs. 2) Fixative-l) containing saturated Digt; 3) After fixation, blocks are washed with phosphate buffer containing 8% sucrose for overnight. 4) Post-fixed in 1% OsO4,(Os) with 0.05M phosphate buffer containing 7% sucrose, at pH 6.8. The blocks stained to deep black(Tann-Protein-Os).

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