Immunoelectron Microscopy of Viral Antigens

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 wi...

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Autores principales: Gulati, Neetu M., Torian, Udana, Gallagher, John R., Harris, Audray K.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6588173/
https://www.ncbi.nlm.nih.gov/pubmed/31219685
http://dx.doi.org/10.1002/cpmc.86
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author Gulati, Neetu M.
Torian, Udana
Gallagher, John R.
Harris, Audray K.
author_facet Gulati, Neetu M.
Torian, Udana
Gallagher, John R.
Harris, Audray K.
author_sort Gulati, Neetu M.
collection PubMed
description 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.
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spelling pubmed-65881732020-06-01 Immunoelectron Microscopy of Viral Antigens Gulati, Neetu M. Torian, Udana Gallagher, John R. Harris, Audray K. Curr Protoc Microbiol Protocol 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. John Wiley and Sons Inc. 2019-06-20 2019-06 /pmc/articles/PMC6588173/ /pubmed/31219685 http://dx.doi.org/10.1002/cpmc.86 Text en © 2019 The Authors. https://creativecommons.org/licenses/by/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Protocol
Gulati, Neetu M.
Torian, Udana
Gallagher, John R.
Harris, Audray K.
Immunoelectron Microscopy of Viral Antigens
title Immunoelectron Microscopy of Viral Antigens
title_full Immunoelectron Microscopy of Viral Antigens
title_fullStr Immunoelectron Microscopy of Viral Antigens
title_full_unstemmed Immunoelectron Microscopy of Viral Antigens
title_short Immunoelectron Microscopy of Viral Antigens
title_sort immunoelectron microscopy of viral antigens
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6588173/
https://www.ncbi.nlm.nih.gov/pubmed/31219685
http://dx.doi.org/10.1002/cpmc.86
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