Label-Free Digital Holo-tomographic Microscopy Reveals Virus-Induced Cytopathic Effects in Live Cells

ABSTRACT Cytopathic effects (CPEs) are a hallmark of infections. CPEs are difficult to observe due to phototoxicity from classical light microscopy. We report distinct patterns of virus infections in live cells using digital holo-tomographic microscopy (DHTM). DHTM is label-free and records the phas...

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Autores principales:	Artur Yakimovich, Robert Witte, Vardan Andriasyan, Fanny Georgi, Urs F. Greber
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2018
Materias:	apoptosis cell contraction cell volume herpes simplex virus label-free microscopy live-cell microscopy Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/d4895b3db8f344e0ae31a6d90c5e5692
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spelling	oai:doaj.org-article:d4895b3db8f344e0ae31a6d90c5e56922021-11-15T15:22:21ZLabel-Free Digital Holo-tomographic Microscopy Reveals Virus-Induced Cytopathic Effects in Live Cells10.1128/mSphereDirect.00599-182379-5042https://doaj.org/article/d4895b3db8f344e0ae31a6d90c5e56922018-12-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphereDirect.00599-18https://doaj.org/toc/2379-5042ABSTRACT Cytopathic effects (CPEs) are a hallmark of infections. CPEs are difficult to observe due to phototoxicity from classical light microscopy. We report distinct patterns of virus infections in live cells using digital holo-tomographic microscopy (DHTM). DHTM is label-free and records the phase shift of low-energy light passing through the specimen on a transparent surface with minimal perturbation. DHTM measures the refractive index (RI) and computes the refractive index gradient (RIG), unveiling optical heterogeneity in cells. We find that vaccinia virus (VACV), herpes simplex virus (HSV), and rhinovirus (RV) infections progressively and distinctly increased RIG. VACV infection, but not HSV and RV infections, induced oscillations of cell volume, while all three viruses altered cytoplasmic membrane dynamics and induced apoptotic features akin to those caused by the chemical compound staurosporine. In sum, we introduce DHTM for quantitative label-free microscopy in infection research and uncover virus type-specific changes and CPE in living cells with minimal interference. IMPORTANCE This study introduces label-free digital holo-tomographic microscopy (DHTM) and refractive index gradient (RIG) measurements of live, virus-infected cells. We use DHTM to describe virus type-specific cytopathic effects, including cyclic volume changes of vaccinia virus infections, and cytoplasmic condensations in herpesvirus and rhinovirus infections, distinct from apoptotic cells. This work shows for the first time that DHTM is suitable to observe virus-infected cells and distinguishes virus type-specific signatures under noninvasive conditions. It provides a basis for future studies, where correlative fluorescence microscopy of cell and virus structures annotate distinct RIG values derived from DHTM.Artur YakimovichRobert WitteVardan AndriasyanFanny GeorgiUrs F. GreberAmerican Society for Microbiologyarticleapoptosiscell contractioncell volumeherpes simplex viruslabel-free microscopylive-cell microscopyMicrobiologyQR1-502ENmSphere, Vol 3, Iss 6 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	apoptosis cell contraction cell volume herpes simplex virus label-free microscopy live-cell microscopy Microbiology QR1-502
spellingShingle	apoptosis cell contraction cell volume herpes simplex virus label-free microscopy live-cell microscopy Microbiology QR1-502 Artur Yakimovich Robert Witte Vardan Andriasyan Fanny Georgi Urs F. Greber Label-Free Digital Holo-tomographic Microscopy Reveals Virus-Induced Cytopathic Effects in Live Cells
description	ABSTRACT Cytopathic effects (CPEs) are a hallmark of infections. CPEs are difficult to observe due to phototoxicity from classical light microscopy. We report distinct patterns of virus infections in live cells using digital holo-tomographic microscopy (DHTM). DHTM is label-free and records the phase shift of low-energy light passing through the specimen on a transparent surface with minimal perturbation. DHTM measures the refractive index (RI) and computes the refractive index gradient (RIG), unveiling optical heterogeneity in cells. We find that vaccinia virus (VACV), herpes simplex virus (HSV), and rhinovirus (RV) infections progressively and distinctly increased RIG. VACV infection, but not HSV and RV infections, induced oscillations of cell volume, while all three viruses altered cytoplasmic membrane dynamics and induced apoptotic features akin to those caused by the chemical compound staurosporine. In sum, we introduce DHTM for quantitative label-free microscopy in infection research and uncover virus type-specific changes and CPE in living cells with minimal interference. IMPORTANCE This study introduces label-free digital holo-tomographic microscopy (DHTM) and refractive index gradient (RIG) measurements of live, virus-infected cells. We use DHTM to describe virus type-specific cytopathic effects, including cyclic volume changes of vaccinia virus infections, and cytoplasmic condensations in herpesvirus and rhinovirus infections, distinct from apoptotic cells. This work shows for the first time that DHTM is suitable to observe virus-infected cells and distinguishes virus type-specific signatures under noninvasive conditions. It provides a basis for future studies, where correlative fluorescence microscopy of cell and virus structures annotate distinct RIG values derived from DHTM.
format	article
author	Artur Yakimovich Robert Witte Vardan Andriasyan Fanny Georgi Urs F. Greber
author_facet	Artur Yakimovich Robert Witte Vardan Andriasyan Fanny Georgi Urs F. Greber
author_sort	Artur Yakimovich
title	Label-Free Digital Holo-tomographic Microscopy Reveals Virus-Induced Cytopathic Effects in Live Cells
title_short	Label-Free Digital Holo-tomographic Microscopy Reveals Virus-Induced Cytopathic Effects in Live Cells
title_full	Label-Free Digital Holo-tomographic Microscopy Reveals Virus-Induced Cytopathic Effects in Live Cells
title_fullStr	Label-Free Digital Holo-tomographic Microscopy Reveals Virus-Induced Cytopathic Effects in Live Cells
title_full_unstemmed	Label-Free Digital Holo-tomographic Microscopy Reveals Virus-Induced Cytopathic Effects in Live Cells
title_sort	label-free digital holo-tomographic microscopy reveals virus-induced cytopathic effects in live cells
publisher	American Society for Microbiology
publishDate	2018
url	https://doaj.org/article/d4895b3db8f344e0ae31a6d90c5e5692
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Label-Free Digital Holo-tomographic Microscopy Reveals Virus-Induced Cytopathic Effects in Live Cells

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