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

Viruses
Nagma ParveenJelle Hendrix

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 microsc...Continue Reading

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Citations

Jan 30, 2021·Wiley Interdisciplinary Reviews. Nanomedicine and Nanobiotechnology·Jun KangJin Chang
Jul 3, 2021·Infectious Disease Reports·Peuli NathAniruddha Ray
Oct 9, 2020·Current Medical Imaging·Alaa A A AljabaliMurtaza M Tambuwala
Jan 18, 2020·Chemical Reviews·Shu-Lin LiuDai-Wen Pang
Nov 2, 2018·Analytical Chemistry·Panagiotis KondylisStephen C Jacobson
Sep 20, 2019·Journal of the American Chemical Society·Nagma ParveenStephan Block

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Methods Mentioned

BETA
fluorescence imaging
fluorescence
confocal microscopy
FRET
protein folding
imaging techniques
super-resolution microscopy
fluorescence correlation spectroscopy
FCS
fluorescence microscopy

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