Retrograde tracing of neural pathways with a protein gold complex. II. Electron microscopic demonstration of projections and collaterals.
Retrograde tracing of neural pathways with a protein gold complex. II. Electron microscopic demonstration of projections and collaterals.
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用蛋白金复合物逆行追踪神经通路。
DOI：
10.1007/bf00492455
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发表时间：
1985
期刊：
影响因子：
--
通讯作者：
Lee,CL
中科院分区：
文献类型：
--
作者：
Menétrey,D;Lee,CL
本文介绍了一种灵敏的方法，用于跟踪神经连接在电子显微镜（EM）水平上使用一种新的化合物，通过耦合的胶体金颗粒的麦胚凝集素辣根过氧化物酶结合物（WGA* HRP-金复合物）。通过金颗粒扫描或银增强后直接在EM水平实现逆行标记细胞的可视化。通过使用不同大小的金颗粒单独耦合到WGA*HRP和注射在不同的脑区EM检测多重逆行标记是可能的。因此，逆行标记的细胞首先通过HRP组织化学与四甲基联苯胺作为色原，然后在电子显微镜下观察渗透和包埋后，在光镜水平上进行鉴定。金颗粒很容易被识别为电子致密，在球形灰色囊泡圆点。不同大小的金颗粒往往定位在同一个囊泡中的识别，建立了蛋白质-金复合物可用于研究父母轴突的抵押。
This paper describes a sensitive method for tracing neural connections at the electron microscopic (EM) level using a new compound produced through the coupling of colloidal gold particles to a wheat germ agglutinin horseradish peroxidase conjugate (the WGA*HRP-gold complex). Visualization of retrogradely labeled cells at the EM level was achieved either directly by gold particles scanning or after silver enhancement. By using different sizes of gold particles individually coupled to WGA*HRP and injected in different brain areas EM detection of multiple retrograde labeling was possible. Thus retrogradely labeled cells were first identified at the light microscopic level through HRP histochemistry with tetramethylbenzidine as a chromogen and then examined under the electron microscope after osmication and embedding. Gold particles were readily identified as electron dense, round dots in spherical grey vesicles. Identification of different sizes of gold particles often localized in the same vesicle established that the protein-gold complex can be used to study collateralisation of parental axons.