免疫细胞化学中的胶体金探针。通过使用银强化程序优化其在光学显微镜中的应用。

Medical biology Pub Date : 1986-01-01
L Scopsi, L I Larsson
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引用次数: 0

摘要

胶体金颗粒是抗原超微结构定位的首选标记物。通过用单宁酸和柠檬酸三钠还原氯化金,可以获得范围广泛的窄定粒度。这样的颗粒可以很容易地偶联到许多蛋白质上,并且所得的偶联物可以方便地在凝胶层析柱上纯化。它们在光学显微镜中的应用需要用银物理显影剂进行放大步骤。银强化胶体金探针可有利地用于低温恒温器、石蜡和塑料切片的免疫染色。此外,渗透培养细胞和全载制剂也可以用金银技术染色。银强化不影响许多组织抗原的反应性,因此允许与免疫过氧化物酶或免疫荧光方法联合进行双重染色。
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Colloidal gold probes in immunocytochemistry. An optimization of their application in light microscopy by use of silver intensification procedures.

Colloidal gold particles are the markers of choice for ultrastructural localization of antigens. By reducing gold chloride with tannic acid and trisodium citrate, a broad range of narrowly determined particle sizes can be obtained. Such particles can easily be coupled to a number of proteins and the resulting conjugates are conveniently purified on a gel-chromatography column. Their application in light microscopy requires an amplification step with a silver physical developer. Silver-intensified colloidal gold probes can advantageously be used for immunostaining of cryostat, paraffin and plastic sections. Moreover, permeabilized cultured cells and whole-mount preparations can also be stained with gold-silver techniques. Silver intensification does not affect reactivity of a number of tissue antigens, thus permitting double staining combinations with immunoperoxidase or immunofluorescence methods.

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