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Application of Advanced Imaging to the Study of Virus Replication

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A special issue of Viruses (ISSN 1999-4915).

Deadline for manuscript submissions: closed (31 December 2017) | Viewed by 39079

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Special Issue Editors

Prof. Dr. Barbara Müller

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Guest Editor

Dept. of Infectious Diseases, Virology, University Hospital Heidelberg, Germany
Interests: Dynamics of HIV-cell interactions; post-entry events in HIV replication; Single Virus Tracing; minimal invasive fluorescent labeling of viral proteins; HIV assembly and maturation

Prof. Dr. Don C. Lamb

E-Mail Website
Guest Editor

Dept. of Physical Chemistry, Ludwig Maximilians University Munich, Munich, Germany
Interests: Ultra-sensitive fluorescence methods, Advanced Microscopy Methods, Live-Cell Imaging, Fluorescence Fluctuation Spectroscopies, Single Molecule Studies, Single Virus Tracing, Protein function and dynamics, Chaperon Assisted Protein Folding, DNA Nanodevices

Special Issue Information

Dear Colleagues,

For decades, electron microscopy has been the method of choice for the visualization of viruses. More recently, novel fluorescence imaging approaches, e.g. super-resolution microscopy, image correlation, and single-molecule techniques, combined with innovative labeling strategies and tracking algorithms, have opened new and exciting possibilities to study the dynamics of virus–host cell interactions and provide insights into the architecture of subviral structures in a complex cellular environment.

This Special Issue of Viruses covers recent advances in virology achieved by employing modern fluorescence imaging techniques. We would like to assemble a collection of primary research papers and reviews that focus on insights in the field of virus–cell or virus–host interaction obtained using advanced fluorescence microscopy or spectroscopy approaches. Topics may include virus imaging, single virus tracing or fluorescence analyses, as well as novel labeling strategies or image analysis methods used to elucidate quantitative and dynamic aspects of virus replication and spread.

Dr. Barbara Müller
Prof. Don C. Lamb
Guest Editors

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Viruses is an international peer-reviewed open access monthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2600 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

Fluorescence microscopy
live-cell imaging
FRET
super-resolution microscopy
fluorescence correlation spectroscopy
image correlation spectroscopy
single-molecule studies

Published Papers (5 papers)

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Research

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12 pages, 1690 KiB

Open AccessArticle

Lipid Composition but Not Curvature Is the Determinant Factor for the Low Molecular Mobility Observed on the Membrane of Virus-Like Vesicles

by Iztok Urbančič, Juliane Brun, Dilip Shrestha, Dominic Waithe, Christian Eggeling and Jakub Chojnacki

Viruses 2018, 10(8), 415; https://doi.org/10.3390/v10080415 - 08 Aug 2018

Cited by 9 | Viewed by 4135

Abstract

Human Immunodeficiency Virus type-1 (HIV-1) acquires its lipid membrane from the plasma membrane of the infected cell from which it buds out. Previous studies have shown that the HIV-1 envelope is an environment of very low mobility, with the diffusion of incorporated proteins two orders of magnitude slower than in the plasma membrane. One of the reasons for this difference is thought to be the HIV-1 membrane composition that is characterised by a high degree of rigidity and lipid packing, which has, until now, been difficult to assess experimentally. To further refine the model of the molecular mobility on the HIV-1 surface, we herein investigated the relative importance of membrane composition and curvature in simplified model membrane systems, large unilamellar vesicles (LUVs) of different lipid compositions and sizes (0.1–1 µm), using super-resolution stimulated emission depletion (STED) microscopy-based fluorescence correlation spectroscopy (STED-FCS). Establishing an approach that is also applicable to measurements of molecule dynamics in virus-sized particles, we found, at least for the 0.1–1 µm sized vesicles, that the lipid composition and thus membrane rigidity, but not the curvature, play an important role in the decreased molecular mobility on the vesicles’ surface. This observation suggests that the composition of the envelope rather than the particle geometry contributes to the previously described low mobility of proteins on the HIV-1 surface. Our vesicle-based study thus provides further insight into the dynamic properties of the surface of individual HIV-1 particles, as well as paves the methodological way towards better characterisation of the properties and function of viral lipid envelopes in general. Full article

(This article belongs to the Special Issue Application of Advanced Imaging to the Study of Virus Replication)

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Review

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22 pages, 856 KiB

Open AccessReview

Visualizing Viral Infection In Vivo by Multi-Photon Intravital Microscopy

by Xaver Sewald

Viruses 2018, 10(6), 337; https://doi.org/10.3390/v10060337 - 20 Jun 2018

Cited by 13 | Viewed by 5902

Abstract

Viral pathogens have adapted to the host organism to exploit the cellular machinery for virus replication and to modulate the host cells for efficient systemic dissemination and immune evasion. Much of our knowledge of the effects that virus infections have on cells originates from in vitro imaging studies using experimental culture systems consisting of cell lines and primary cells. Recently, intravital microscopy using multi-photon excitation of fluorophores has been applied to observe virus dissemination and pathogenesis in real-time under physiological conditions in living organisms. Critical steps during viral infection and pathogenesis could be studied by direct visualization of fluorescent virus particles, virus-infected cells, and the immune response to viral infection. In this review, I summarize the latest research on in vivo studies of viral infections using multi-photon intravital microscopy (MP-IVM). Initially, the underlying principle of multi-photon microscopy is introduced and experimental challenges during microsurgical animal preparation and fluorescent labeling strategies for intravital imaging are discussed. I will further highlight recent studies that combine MP-IVM with optogenetic tools and transcriptional analysis as a powerful approach to extend the significance of in vivo imaging studies of viral pathogens. Full article

(This article belongs to the Special Issue Application of Advanced Imaging to the Study of Virus Replication)

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15 pages, 1720 KiB

Open AccessReview

Live-Cell Imaging of Early Steps of Single HIV-1 Infection

by Ashwanth C. Francis and Gregory B. Melikyan

Viruses 2018, 10(5), 275; https://doi.org/10.3390/v10050275 - 19 May 2018

Cited by 26 | Viewed by 6546

Abstract

Live-cell imaging of single HIV-1 entry offers a unique opportunity to delineate the spatio-temporal regulation of infection. Novel virus labeling and imaging approaches enable the visualization of key steps of HIV-1 entry leading to nuclear import, integration into the host genome, and viral protein expression. Here, we discuss single virus imaging strategies, focusing on live-cell imaging of single virus fusion and productive uncoating that culminates in HIV-1 infection. Full article

(This article belongs to the Special Issue Application of Advanced Imaging to the Study of Virus Replication)

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21 pages, 2798 KiB

Open AccessReview

Single Viruses on the Fluorescence Microscope: Imaging Molecular Mobility, Interactions and Structure Sheds New Light on Viral Replication

by Nagma Parveen, Doortje Borrenberghs, Susana Rocha and Jelle Hendrix

Viruses 2018, 10(5), 250; https://doi.org/10.3390/v10050250 - 10 May 2018

Cited by 20 | Viewed by 8011

Abstract

Viruses are simple agents exhibiting complex reproductive mechanisms. Decades of research have provided crucial basic insights, antiviral medication and moderately successful gene therapy trials. The most infectious viral particle is, however, not always the most abundant one in a population, questioning the utility of classic ensemble-averaging virology. Indeed, viral replication is often not particularly efficient, prone to errors or containing parallel routes. Here, we review different single-molecule sensitive fluorescence methods that we employ routinely to investigate viruses. We provide a brief overview of the microscopy hardware needed and discuss the different methods and their application. In particular, we review how we applied (i) single-molecule Förster resonance energy transfer (smFRET) to probe the subviral human immunodeficiency virus (HIV-1) integrase (IN) quaternary structure; (ii) single particle tracking to study interactions of the simian virus 40 with membranes; (iii) 3D confocal microscopy and smFRET to quantify the HIV-1 pre-integration complex content and quaternary structure; (iv) image correlation spectroscopy to quantify the cytosolic HIV-1 Gag assembly, and finally; (v) super-resolution microscopy to characterize the interaction of HIV-1 with tetherin during assembly. We hope this review is an incentive for setting up and applying similar single-virus imaging studies in daily virology practice. Full article

(This article belongs to the Special Issue Application of Advanced Imaging to the Study of Virus Replication)

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31 pages, 936 KiB

Open AccessReview

Concepts in Light Microscopy of Viruses

by Robert Witte, Vardan Andriasyan, Fanny Georgi, Artur Yakimovich and Urs F. Greber

Viruses 2018, 10(4), 202; https://doi.org/10.3390/v10040202 - 18 Apr 2018

Cited by 36 | Viewed by 13549

Abstract

Viruses threaten humans, livestock, and plants, and are difficult to combat. Imaging of viruses by light microscopy is key to uncover the nature of known and emerging viruses in the quest for finding new ways to treat viral disease and deepening the understanding of virus–host interactions. Here, we provide an overview of recent technology for imaging cells and viruses by light microscopy, in particular fluorescence microscopy in static and live-cell modes. The review lays out guidelines for how novel fluorescent chemical probes and proteins can be used in light microscopy to illuminate cells, and how they can be used to study virus infections. We discuss advantages and opportunities of confocal and multi-photon microscopy, selective plane illumination microscopy, and super-resolution microscopy. We emphasize the prevalent concepts in image processing and data analyses, and provide an outlook into label-free digital holographic microscopy for virus research. Full article

(This article belongs to the Special Issue Application of Advanced Imaging to the Study of Virus Replication)

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