Wouterlood F G
Stain Technol. 1986 Nov;61(6):337-52. doi: 10.3109/10520298609113582.
Deterioration of Golgi impregnation begins immediately after impregnated tissue blocks are sectioned with the Vibratome. The first signs of deterioration are fading of delicate impregnated processes, the disruption and fragmentation of dendrites, and, eventually, fading of entire neurons. These changes can be prevented by stabilization, i.e., by converting the water soluble silver chromate Golgi precipitate into metallic silver or by replacing the silver with some other dense, insoluble material. A technique is described using photographic developers to treat Vibratome sections containing Golgi-rapid or Golgi-Kopsch impregnated CNS neurons. In this way part of the silver chromate Golgi precipitate is reduced to metallic silver, and the remaining silver chromate is then removed with sodium thiosulfate. Of the various developers tested, Kodalith and Elon-ascorbic acid gave the best results, with excellent stabilization of the most delicate structures, such as the stalks of dendritic spines and finely woven axonal plexuses. Treatment with other developers (HC-110, Neutol, D-19, D-76, D-163, Kodak Universal, Rodinal, Atomal, Diafine, Eukobrom, Microdol-X) resulted in stabilization ranging from good to poor. Good stabilization of Golgi impregnation could also be achieved by first exposing the sections to sodium bromide (bromide substitution) followed by treatment with D-19, Kodalith, Elon-ascorbic acid or HC-110. After stabilization, the sections can be counterstained with aqueous cresyl violet or with alcoholic thionin without degradation of the stabilized Golgi image. The counterstain permits exact determination of the position of impregnated neurons in cortical layers or subcortical nuclei.
在用振动切片机对浸银组织块进行切片后,高尔基染色法的染色效果会立即开始变差。染色变差的最初迹象是精细的染色突起褪色、树突断裂和破碎,最终整个神经元褪色。这些变化可以通过稳定化处理来防止,即通过将水溶性的铬酸银高尔基沉淀物转化为金属银,或者用其他一些致密、不溶性物质替代银。本文描述了一种使用显影剂处理含有高尔基快速法或高尔基-科普施法染色的中枢神经系统神经元的振动切片机切片的技术。通过这种方法,部分铬酸银高尔基沉淀物被还原为金属银,然后用硫代硫酸钠去除剩余的铬酸银。在测试的各种显影剂中,柯达立石显影剂和埃隆-抗坏血酸组合效果最佳,能极好地稳定最精细的结构,如树突棘的柄和精细交织的轴突丛。用其他显影剂(HC-110、纽托尔、D-19、D-76、D-163、柯达通用显影剂、罗丹诺、阿托马尔、迪亚芬、优可溴、微粒显影剂-X)处理后,稳定化效果从好到差不等。通过先将切片暴露于溴化钠(溴替代),然后用D-19、柯达立石显影剂、埃隆-抗坏血酸或HC-110处理,也能实现高尔基染色的良好稳定化。稳定化处理后,切片可以用水溶性甲酚紫或酒精性硫堇进行复染,而不会破坏稳定化的高尔基染色图像。复染可以准确确定染色神经元在皮质层或皮质下核中的位置。