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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...

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Detalles Bibliográficos
Autores principales: Pedersen, Meike, Jamali, Shirin, Saha, Ipsita, Daum, Rainer, Bendjennat, Mourad, Saffarian, Saveez
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer International Publishing 2018
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.
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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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