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{"title":"Immunoelectron Microscopy of Viral Antigens","authors":"Neetu M. Gulati, Udana Torian, John R. Gallagher, Audray K. Harris","doi":"10.1002/cpmc.86","DOIUrl":null,"url":null,"abstract":"<p>Immunoelectron microscopy is a powerful technique for identifying viral antigens and determining their structural localization and organization within vaccines and viruses. While traditional negative staining transmission electron microscopy provides structural information, identity of components within a sample may be confounding. Immunoelectron microscopy allows for identification and visualization of antigens and their relative positions within a particulate sample. This allows for simple qualitative analysis of samples including whole virus, viral components, and viral-like particles. This article describes methods for immunogold labeling of viral antigens in a liquid suspension, with examples of immunogold-labeled influenza virus glycoproteins, and also discusses the important considerations for sample preparation and determination of morphologies. Together, these methods allow for understanding the antigenic makeup of viral particulate samples, which have important implications for molecular virology and vaccine development. © 2019 The Authors. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.</p>","PeriodicalId":39967,"journal":{"name":"Current Protocols in Microbiology","volume":"53 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2019-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpmc.86","citationCount":"12","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols in Microbiology","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cpmc.86","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
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Abstract
Immunoelectron microscopy is a powerful technique for identifying viral antigens and determining their structural localization and organization within vaccines and viruses. While traditional negative staining transmission electron microscopy provides structural information, identity of components within a sample may be confounding. Immunoelectron microscopy allows for identification and visualization of antigens and their relative positions within a particulate sample. This allows for simple qualitative analysis of samples including whole virus, viral components, and viral-like particles. This article describes methods for immunogold labeling of viral antigens in a liquid suspension, with examples of immunogold-labeled influenza virus glycoproteins, and also discusses the important considerations for sample preparation and determination of morphologies. Together, these methods allow for understanding the antigenic makeup of viral particulate samples, which have important implications for molecular virology and vaccine development. © 2019 The Authors. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
病毒抗原的免疫电子显微镜
免疫电子显微镜是一种识别病毒抗原并确定其在疫苗和病毒中的结构定位和组织的强大技术。虽然传统的负染色透射电子显微镜提供了结构信息,但样品中成分的身份可能会混淆。免疫电子显微镜可以识别和可视化抗原及其在颗粒样品中的相对位置。这允许对样品进行简单的定性分析,包括整个病毒、病毒成分和病毒样颗粒。本文介绍了在液体悬浮液中对病毒抗原进行免疫金标记的方法,以免疫金标记流感病毒糖蛋白为例,并讨论了样品制备和形态学测定的重要注意事项。总之,这些方法允许了解病毒颗粒样品的抗原组成,这对分子病毒学和疫苗开发具有重要意义。©2019作者。这是一篇基于知识共享署名许可协议的开放获取文章,该协议允许在任何媒体上使用、分发和复制,前提是正确引用原始作品。
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