Marfurt C F, Turner D F, Adams C E
Department of Anatomy, Northwest Center for Medical Education, Indiana University School of Medicine, Gary 46408.
J Neurosci Methods. 1988 Oct;25(3):215-23. doi: 10.1016/0165-0270(88)90136-7.
The ability to use the tetramethylbenzidine (TMB) method for studying neuronal connections at the electron microscopic level is often difficult because the conditions of osmification and dehydration used in processing the tissue may result in significant loss and/or decreased electron density of the reaction product. In the present study, we report that stabilization of TMB reaction product with 5% ammonium molybdate (AM) prior to osmificating the tissue results in the formation of TMB-AM crystals that are many times more electron dense and resistant to ethanol extraction than non-stabilized TMB crystals. The nature of the chemical interaction that underlies the stabilization of TMB by AM is uncertain, but it may involve the formation of an insoluble salt between molybdic ions and the TMB polymer. The use of this simple procedure increases the sensitivity of the TMB procedure at the electron microscopic level and may be used to label neuronal pathways in the peripheral and central nervous systems with equal success.
在电子显微镜水平上使用四甲基联苯胺（TMB）方法研究神经元连接的能力往往存在困难，因为处理组织时所采用的锇化和脱水条件可能会导致反应产物显著损失和/或电子密度降低。在本研究中，我们报告称，在组织锇化之前用5%钼酸铵（AM）稳定TMB反应产物会导致形成TMB-AM晶体，其电子密度比未稳定的TMB晶体高许多倍，并且对乙醇提取具有抗性。AM稳定TMB背后的化学相互作用的本质尚不确定，但可能涉及钼离子与TMB聚合物之间形成不溶性盐。使用这个简单的程序可提高TMB程序在电子显微镜水平上的灵敏度，并且可同等成功地用于标记外周和中枢神经系统中的神经元通路。
