Lemann W, Saper C B, Rye D B, Wainer B H
Brain Res Bull. 1985 Mar;14(3):277-81. doi: 10.1016/0361-9230(85)90093-0.
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).
用于电子显微镜的辣根过氧化物酶组织化学的高灵敏度四甲基联苯胺（TMB）方法，一直受到反应产物在水溶液和醇溶液中溶解度的限制。我们发现，在TMB反应后进行二氨基联苯胺-钴（DAB-Co）步骤会使TMB晶体被DAB-Co包裹。所得的反应复合物不溶，并且使用电子显微镜很容易定位。通过系统地改变进行TMB反应时的pH值，可以控制反应复合物的大小和形状。pH 4.0的反应复合物最适合用于电子显微镜鉴定直径小于1.0微米的标记结构（例如轴突终末）。
