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Correlative iPALM and SEM resolves virus cavity and Gag lattice defects in HIV virions
Interferometric Photo-Activation-Localization-Microscopy (iPALM) localizes single fluorescent molecules with 20 nm lateral and 10 nm axial resolution. We present a method utilizing glass coverslip lithography for correlative imaging between iPALM and scanning electron microscopy (SEM). Using iPALM o...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer International Publishing
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6330563/ https://www.ncbi.nlm.nih.gov/pubmed/30043246 http://dx.doi.org/10.1007/s00249-018-1324-0 |
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author | Pedersen, Meike Jamali, Shirin Saha, Ipsita Daum, Rainer Bendjennat, Mourad Saffarian, Saveez |
author_facet | Pedersen, Meike Jamali, Shirin Saha, Ipsita Daum, Rainer Bendjennat, Mourad Saffarian, Saveez |
author_sort | Pedersen, Meike |
collection | PubMed |
description | Interferometric Photo-Activation-Localization-Microscopy (iPALM) localizes single fluorescent molecules with 20 nm lateral and 10 nm axial resolution. We present a method utilizing glass coverslip lithography for correlative imaging between iPALM and scanning electron microscopy (SEM). Using iPALM on HIV Gag-Dendra virus-like particles (VLPs) we localized the position of HIV Gag proteins. Based on these localizations we reconstructed the central cavity of the VLPs along with imperfections within the HIV Gag lattice. The SEM images and iPALM images overlap and show imaging from single VLPs immobilized on glass coverslips. The localization of many HIV proteins including accessory proteins and Gag-Pol remains unknown, we discuss how the specificity of iPALM coupled with SEM has the potential for resolving more of HIV proteins. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s00249-018-1324-0) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-6330563 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Springer International Publishing |
record_format | MEDLINE/PubMed |
spelling | pubmed-63305632019-01-27 Correlative iPALM and SEM resolves virus cavity and Gag lattice defects in HIV virions Pedersen, Meike Jamali, Shirin Saha, Ipsita Daum, Rainer Bendjennat, Mourad Saffarian, Saveez Eur Biophys J Original Article Interferometric Photo-Activation-Localization-Microscopy (iPALM) localizes single fluorescent molecules with 20 nm lateral and 10 nm axial resolution. We present a method utilizing glass coverslip lithography for correlative imaging between iPALM and scanning electron microscopy (SEM). Using iPALM on HIV Gag-Dendra virus-like particles (VLPs) we localized the position of HIV Gag proteins. Based on these localizations we reconstructed the central cavity of the VLPs along with imperfections within the HIV Gag lattice. The SEM images and iPALM images overlap and show imaging from single VLPs immobilized on glass coverslips. The localization of many HIV proteins including accessory proteins and Gag-Pol remains unknown, we discuss how the specificity of iPALM coupled with SEM has the potential for resolving more of HIV proteins. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s00249-018-1324-0) contains supplementary material, which is available to authorized users. Springer International Publishing 2018-07-24 2019 /pmc/articles/PMC6330563/ /pubmed/30043246 http://dx.doi.org/10.1007/s00249-018-1324-0 Text en © The Author(s) 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. |
spellingShingle | Original Article Pedersen, Meike Jamali, Shirin Saha, Ipsita Daum, Rainer Bendjennat, Mourad Saffarian, Saveez Correlative iPALM and SEM resolves virus cavity and Gag lattice defects in HIV virions |
title | Correlative iPALM and SEM resolves virus cavity and Gag lattice defects in HIV virions |
title_full | Correlative iPALM and SEM resolves virus cavity and Gag lattice defects in HIV virions |
title_fullStr | Correlative iPALM and SEM resolves virus cavity and Gag lattice defects in HIV virions |
title_full_unstemmed | Correlative iPALM and SEM resolves virus cavity and Gag lattice defects in HIV virions |
title_short | Correlative iPALM and SEM resolves virus cavity and Gag lattice defects in HIV virions |
title_sort | correlative ipalm and sem resolves virus cavity and gag lattice defects in hiv virions |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6330563/ https://www.ncbi.nlm.nih.gov/pubmed/30043246 http://dx.doi.org/10.1007/s00249-018-1324-0 |
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