A simplified procedure for the physical development of the sulphide silver method to reveal synaptic zinc in combination with immunocytochemistry at light and electron microscopy
A simplified procedure for the physical development of the sulphide silver method to reveal synaptic zinc in combination with immunocytochemistry at light and electron microscopy
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DOI：
10.1016/s0165-0270(97)00169-6
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发表时间：
1998-01-31
影响因子：
3
通讯作者：
Bendotti, C
中科院分区：
文献类型：
--
作者：
De Biasi, S;Bendotti, C
正常和病理条件下（例如红藻氨酸诱导的癫痫持续状态）中兴奋性突触末梢中存在的锌池可以通过硫化银方法染色，然后物理形成不溶性的硫化锌复合物。在这项研究中，我们将先前描述的揭示突触锌的简单且快速的开发程序应用于正常和病理海马的研究，并将其与嵌入前和嵌入后免疫细胞化学方法相结合以检测不同的抗原。用含有硫化钠的固定液灌注正常大鼠和红藻氨酸处理的大鼠，并将50μm厚的海马振动切片在商业显影液(IntenSE M，Amersham)中孵育。然后，将开发的振动切片机切片（1）安装用于光学显微镜检查或锇化和 epon 嵌入用于电子显微镜检查； (2) 进行处理，用于通过光学和电子显微镜对各种抗原（GABA、小白蛋白、钙结合蛋白）进行预包埋免疫酶检测。来自 epon 包埋样品的薄切片也经过处理，用于谷氨酸的包埋后免疫金定位。这种非常简单和快速的过程产生了锌特异性染色，与经典显色方法获得的染色相当，并且抗原性和超微结构都得到了良好的保存。因此可以在相同的厚切片或薄切片中检测锌反应产物和不同的抗原。 (C) 1998 Elsevier Science B.V.
The pool of zinc present in excitatory synaptic terminals in normal and pathological conditions (for instance the status epilepticus induced by kainic acid) can be stained by a silver sulphide method followed by physical development of the insoluble zinc-sulphide complexes. In this study we applied a previously described simple and rapid developing procedure that reveals synaptic zinc, to the study of normal and pathological hippocampi and combined it with pre and postembedding immunocytochemical methods to detect different antigens. Normal and kainic acid-treated rats were perfused with fixative solutions containing sodium sulphide and 50 mu m-thick vibratome sections of the hippocampi were incubated in a commercial developing solution (IntenSE M, Amersham). The developed vibratome sections were then (1) mounted for light microscopy or osmicated and epon-embedded for electron microscopy; or (2) processed for the preembedding immunoenzymatic detection of various antigens (GABA, parvalbumin, calbindin) with light and electron microscopy. Thin sections from epon-embedded samples were also processed for the postembedding immunogold localization of glutamate. This very simple and rapid procedure gives rise to zinc-specific staining, comparable to that obtained with classical developing methods and good preservation of both antigenicity and ultrastructure. It is therefore possible to detect, in the same thick or thin section, zinc reaction product and different antigens. (C) 1998 Elsevier Science B.V.