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. 2020 Dec 14;10(72):44079-44086.
doi: 10.1039/d0ra09083b. eCollection 2020 Dec 9.

Interpretation of SARS-CoV-2 behaviour on different substrates and denaturation of virions using ethanol: an atomic force microscopy study

Umit Celik  1 , Kubra Celik  1 , Suleyman Celik  2 , Hasan Abayli  3 , Kezban Can Sahna  3 , Şükrü Tonbak  3 , Zulal Asci Toraman  4 , Ahmet Oral  5   6
Affiliations

Interpretation of SARS-CoV-2 behaviour on different substrates and denaturation of virions using ethanol: an atomic force microscopy study

Umit Celik et al. RSC Adv. .

Abstract

Coronavirus (SARS-CoV-2) is a respiratory infection virus that was first detected in Wuhan, China. The virus causes COVID-19 disease and the outbreak was recognised as a pandemic by the World Health Organization (WHO) in March 2020. SARS-CoV-2 virion was first imaged using cryo-electron microscopy by the Chinese Center for Disease Control and Prevention (CDC). Atomic Force Microscopy is a unique technique that can allow imaging of biomolecules under different conditions. In this work, we used Atomic Force Microscopy to characterize SARS-CoV-2 on tissue culture polystyrene (TCPS) and glass coverslip surfaces. We isolated SARS-CoV-2 and drop casted it on coverslip glass and tissue culture polystyrene surfaces. We analyzed height profiles, density, and aggregation behavior of the virion on glass and polystyrene surfaces. We observed the coffee ring effect on the drop casted samples and close packing of virions near the coffee rings on both surfaces with relatively higher virion distribution on the tissue culture polystyrene (TCPS) substrates. We compare virion agglomeration on the two types of surfaces. Finally, we applied ethanol disinfectant to virions on the surface to visualize the effect of ethanol and image the ultrastructure of SARS-CoV-2.

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Conflict of interest statement

There are no conflicts to declare.

Figures

Fig. 1
Fig. 1. Simplified colour coded illustration model for SARS-CoV-2 virus. The gray surface is a spherical bilayer lipid envelope. The more abundant membrane or M proteins are shown in orange. The envelope or E membranes are shown in yellow and the spikey S proteins are shown in red. Image courtesy CDC/Alisa Eckert and Dan Higgins.
Fig. 2
Fig. 2. AFM images of SARS-CoV-2. (A) Topography image, (B) corresponding phase image.
Fig. 3
Fig. 3. (A) Illustration of nanoshaving, (B) SARS-CoV-2 AFM topography image after nanoshaving on the glass substrate.
Fig. 4
Fig. 4. SARS-CoV-2 AFM topography images. (A) Imaged on TCPS substrate, (B) imaged on glass substrate.
Fig. 5
Fig. 5. Distribution of the maximal central height of SARS-CoV-2 virions histograms on TCPS and glass substrates obtained from inset AFM topography images.
Fig. 6
Fig. 6. Agglomeration behaviour of SARS-CoV-2. AFM topography images, (A) on glass substrate, (B) on TCPS substrate.
Fig. 7
Fig. 7. TCPS substrate AFM topography image, (A) before deposition of virion solution, (B) after deposition of virion solution.
Fig. 8
Fig. 8. An illustration of distribution of virions on the substrates.
Fig. 9
Fig. 9. AFM images closed packed SARS-CoV-2 on glass substrate, (A) topography image, 5 × 5 μm. Inset shows the optical image of coffee ring and scanned area marked as in red. (B) Zoom in topography image of (A). Virion diameters measured as 107 ± 15 nm.
Fig. 10
Fig. 10. AFM topography images of SARS-CoV-2 on TCPS substrate, (A) before exposing to 80% ethyl alcohol, 5 × 5 μm2. Inset shows zoom in image of virus multimers. (B) Surface after exposing the specimen to 80% ethyl alcohol.
Fig. 11
Fig. 11. AFM images of SARS-CoV-2 on TCPS substrate, (A) topography image after exposing to 80% ethyl alcohol, (B) corresponding phase image of (A), (C) illustration of 80% ethyl alcohol effect on SARS-CoV-2.
Fig. 12
Fig. 12. AFM Topography image of SARS-CoV-2 on glass substrate after exposing to 80% ethyl alcohol. (A) AFM topography image (B) 3D topography image of (A).
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