Light Sheet Microscopy-Assisted 3D Analysis of SARS-CoV-2 Infection in the Respiratory Tract of the Ferret Model
The visualization of viral pathogens in infected tissues is an invaluable tool to understand spatial virus distribution, localization, and cell tropism in vivo. Commonly, virus-infected tissues are analyzed using conventional immunohistochemistry in paraffin-embedded thin sections. Here, we demonstr...
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-03-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/13/3/529 |
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| author | Luca M. Zaeck David Scheibner Julia Sehl Martin Müller Donata Hoffmann Martin Beer Elsayed M. Abdelwhab Thomas C. Mettenleiter Angele Breithaupt Stefan Finke |
| author_facet | Luca M. Zaeck David Scheibner Julia Sehl Martin Müller Donata Hoffmann Martin Beer Elsayed M. Abdelwhab Thomas C. Mettenleiter Angele Breithaupt Stefan Finke |
| author_sort | Luca M. Zaeck |
| collection | DOAJ |
| description | The visualization of viral pathogens in infected tissues is an invaluable tool to understand spatial virus distribution, localization, and cell tropism in vivo. Commonly, virus-infected tissues are analyzed using conventional immunohistochemistry in paraffin-embedded thin sections. Here, we demonstrate the utility of volumetric three-dimensional (3D) immunofluorescence imaging using tissue optical clearing and light sheet microscopy to investigate host–pathogen interactions of pandemic SARS-CoV-2 in ferrets at a mesoscopic scale. The superior spatial context of large, intact samples (>150 mm<sup>3</sup>) allowed detailed quantification of interrelated parameters like focus-to-focus distance or SARS-CoV-2-infected area, facilitating an in-depth description of SARS-CoV-2 infection foci. Accordingly, we could confirm a preferential infection of the ferret upper respiratory tract by SARS-CoV-2 and suggest clustering of infection foci in close proximity. Conclusively, we present a proof-of-concept study for investigating critically important respiratory pathogens in their spatial tissue morphology and demonstrate the first specific 3D visualization of SARS-CoV-2 infection. |
| first_indexed | 2024-03-10T12:57:50Z |
| format | Article |
| id | doaj.art-2a981cd7d8914797bc7d0f1bad3f1350 |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-10T12:57:50Z |
| publishDate | 2021-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-2a981cd7d8914797bc7d0f1bad3f13502023-11-21T11:43:40ZengMDPI AGViruses1999-49152021-03-0113352910.3390/v13030529Light Sheet Microscopy-Assisted 3D Analysis of SARS-CoV-2 Infection in the Respiratory Tract of the Ferret ModelLuca M. Zaeck0David Scheibner1Julia Sehl2Martin Müller3Donata Hoffmann4Martin Beer5Elsayed M. Abdelwhab6Thomas C. Mettenleiter7Angele Breithaupt8Stefan Finke9Institute of Molecular Virology and Cell Biology, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, 17493 Greifswald-Insel Riems, GermanyInstitute of Molecular Virology and Cell Biology, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, 17493 Greifswald-Insel Riems, GermanyDepartment of Experimental Animal Facilities and Biorisk Management, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, 17493 Greifswald-Insel Riems, GermanyInstitute of Molecular Virology and Cell Biology, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, 17493 Greifswald-Insel Riems, GermanyInstitute of Diagnostic Virology, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, 17493 Greifswald-Insel Riems, GermanyInstitute of Diagnostic Virology, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, 17493 Greifswald-Insel Riems, GermanyInstitute of Molecular Virology and Cell Biology, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, 17493 Greifswald-Insel Riems, GermanyFriedrich-Loeffler-Institut, Federal Research Institute for Animal Health, 17493 Greifswald-Insel Riems, GermanyDepartment of Experimental Animal Facilities and Biorisk Management, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, 17493 Greifswald-Insel Riems, GermanyInstitute of Molecular Virology and Cell Biology, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, 17493 Greifswald-Insel Riems, GermanyThe visualization of viral pathogens in infected tissues is an invaluable tool to understand spatial virus distribution, localization, and cell tropism in vivo. Commonly, virus-infected tissues are analyzed using conventional immunohistochemistry in paraffin-embedded thin sections. Here, we demonstrate the utility of volumetric three-dimensional (3D) immunofluorescence imaging using tissue optical clearing and light sheet microscopy to investigate host–pathogen interactions of pandemic SARS-CoV-2 in ferrets at a mesoscopic scale. The superior spatial context of large, intact samples (>150 mm<sup>3</sup>) allowed detailed quantification of interrelated parameters like focus-to-focus distance or SARS-CoV-2-infected area, facilitating an in-depth description of SARS-CoV-2 infection foci. Accordingly, we could confirm a preferential infection of the ferret upper respiratory tract by SARS-CoV-2 and suggest clustering of infection foci in close proximity. Conclusively, we present a proof-of-concept study for investigating critically important respiratory pathogens in their spatial tissue morphology and demonstrate the first specific 3D visualization of SARS-CoV-2 infection.https://www.mdpi.com/1999-4915/13/3/529SARS-CoV-2COVID-193D immunofluorescencelight sheet microscopyconfocal laser scanning microscopytissue optical clearing |
| spellingShingle | Luca M. Zaeck David Scheibner Julia Sehl Martin Müller Donata Hoffmann Martin Beer Elsayed M. Abdelwhab Thomas C. Mettenleiter Angele Breithaupt Stefan Finke Light Sheet Microscopy-Assisted 3D Analysis of SARS-CoV-2 Infection in the Respiratory Tract of the Ferret Model Viruses SARS-CoV-2 COVID-19 3D immunofluorescence light sheet microscopy confocal laser scanning microscopy tissue optical clearing |
| title | Light Sheet Microscopy-Assisted 3D Analysis of SARS-CoV-2 Infection in the Respiratory Tract of the Ferret Model |
| title_full | Light Sheet Microscopy-Assisted 3D Analysis of SARS-CoV-2 Infection in the Respiratory Tract of the Ferret Model |
| title_fullStr | Light Sheet Microscopy-Assisted 3D Analysis of SARS-CoV-2 Infection in the Respiratory Tract of the Ferret Model |
| title_full_unstemmed | Light Sheet Microscopy-Assisted 3D Analysis of SARS-CoV-2 Infection in the Respiratory Tract of the Ferret Model |
| title_short | Light Sheet Microscopy-Assisted 3D Analysis of SARS-CoV-2 Infection in the Respiratory Tract of the Ferret Model |
| title_sort | light sheet microscopy assisted 3d analysis of sars cov 2 infection in the respiratory tract of the ferret model |
| topic | SARS-CoV-2 COVID-19 3D immunofluorescence light sheet microscopy confocal laser scanning microscopy tissue optical clearing |
| url | https://www.mdpi.com/1999-4915/13/3/529 |
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