Stabilization of TMB Reaction Product for electron microscopic retrograde and anterograde fiber tracing
Brain Research BulletinPublished 1 March 1985
Walter Lemann, Clifford B. Saper, David B. Rye, Bruce H. Wainer
Citations73
SJR quartileQ2
SJR score1.07
SNIP0.82
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TL;DR
It is found that following the TMB reaction with a diaminobenzidine-cobalt (DAB-Co) step causes the T MB crystals to become coated with DAB-Co, and the resultant reaction complex is insoluble, and easily localized using electron microscopy.
Abstract
Use of the highly sensitive tetramethylbenzidine (TMB) method of horseradish peroxidase histochemistry for electron microscopy has been limited by the solubility of the reaction product in aqueous and alcoholic solutions. We have found that following the TMB reaction with a diaminobenzidine-cobalt (DAB-Co) step causes the TMB crystals to become coated with DAB-Co. The resultant reaction complex is insoluble, and easily localized using electron microscopy. By systematically varying the pH at which the TMB reaction is run, the size and shape of the reaction complex can be controlled. The pH 4.0 reaction complex was the most suitable for electron microscopic identification of labeled structures less than 1.0 micron in diameter (e.g., axon terminals).
Keywords
Biochemistry, Genetics and Molecular Biology
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