[以白色念珠菌表面抗原蛋白为例,比较扫描电镜下各种免疫表面标记方法]。

M Borg
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引用次数: 0

摘要

免疫复合物的扫描电子显微镜(SEM)标记是一项相当新的技术,各种程序,已提出,尚未比较。以念珠菌蛋白酶为靶抗原进行比较评价。这种条件酵母菌白色念珠菌的分泌酶在蛋白培养基中生长和感染小鼠腹膜巨噬细胞后,都能定位于真菌囊胚孔和菌丝表面。通过免疫荧光和免疫过氧化物酶光显微镜证实了蛋白酶抗原的存在。用免疫过氧化物酶反应产生的胶体、免疫金技术和与合成聚合物(聚苯乙烯、聚甲基丙烯酸酯、聚丙烯醛)连接的抗体,对蛋白酶-抗蛋白酶复合物的SEM进行了修饰。此外,还使用了灭活的金黄色葡萄球菌,它通过其蛋白a与抗体结合。表面结构的扫描电镜高分辨率是胶体修饰技术所不能比拟的。所有与小珠结合的偶联物都存在不一致的结合,这与表面抗原的分布不一致。用聚苯乙烯(乳胶)制备微球效果较好。此外,基于胶体的技术允许临界点干燥,这不能以通常的方式应用于合成珠。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
[Comparison of various immune surface labeling methods for scanning electron microscopy with the example of a surface antigen protein of the yeast Candida albicans].

The labeling of immunocomplexes for scanning electron microscopy (SEM) is a fairly new technique, and the various procedures, that have been proposed, have not yet been compared. Such comparative evaluation was performed with Candida protease as a target antigen. This secretory enzyme of the opportunistic yeast Candida albicans can be localized on the surface of fungal blastopores and mycelia, both after growth in proteinaceous medium and upon infection of murine peritoneal macrophages. The presence of the protease antigen was confirmed by immunofluorescence and by immunoperoxidase-light microscopy. The decoration of protease - anti protease complexes for SEM was attempted with colloids derived from the immunoperoxidase reaction, by the immunogold technique, and by antibodies linked to beads of synthetic polymers (polystyrene, polymethacrylate, polyacrolein). In addition, inactivated Staphylococcus aureus was used, which binds to antibodies through its protein-A. The high resolution by SEM of surface structures was matched only by the colloid based decoration techniques. All conjugates with beads suffered from inconsistent binding, which did not correspond with the distribution of the surface antigen. The comparatively best result with beads was obtained with polystyrene (Latex). Colloid based techniques in addition allow for critical point drying, which cannot be applied to synthetic beads in the usual manner.

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