Atomic force microscopy analysis of native infectious and inactivated SARS-CoV-2 virions

AbstractSARS-CoV-2 is an enveloped virus responsible for the Coronavirus Disease 2019 (COVID-19) pandemic. Here, single viruses were analyzed by atomic force microscopy (AFM) operating directly in a level 3 biosafety (BSL3) facility, which appeared as a fast and powerful method to assess at the nanoscale level and in 3D infectious virus morphology in its native conformation, or upon inactivation treatments. AFM imaging reveals structurally intact infectious and inactivated SARS-CoV-2 upon low concentration of formaldehyde treatment. This protocol combining AFM and plaque assays allows the preparation of intact inactivated SARS-CoV-2 particles for safe use of samples out of level 3 laboratory to accelerate researches against the COVID-19 pandemic. Overall, we illustrate how adapted BSL3-AFM is a remarkable toolbox for rapid and direct virus analysis based on nanoscale morphology.

Download Full-text

Direct visualization of native infectious SARS-CoV-2 and its inactivation forms using high resolution Atomic Force Microscopy

10.1101/2020.10.23.351916 ◽

2020 ◽

Author(s):

Sébastien Lyonnais ◽

Mathilde Hénaut ◽

Aymeric Neyret ◽

Peggy Merida ◽

Chantal Cazevieille ◽

...

Keyword(s):

Atomic Force Microscopy ◽

High Resolution ◽

Native Conformation ◽

Virus Identification ◽

Force Microscopy ◽

Atomic Force ◽

Level 3 ◽

Identification And Characterization ◽

Virus Morphology ◽

Nanoscale Level

AbstractThe severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is responsible for COVID19, a new emerging pandemic affecting humans. Here, single viruses were analyze by atomic force microscopy (AFM) operating directly in a level 3 biosafety (BSL3) facility, which appeared as a fast and powerful method to assess infectious virus morphology in its native conformation, or upon inactivation treatments, at the nanoscale level and in 3D. High resolution AFM reveals structurally intact infectious and inactivated SARS-CoV-2 upon low concentration of formaldehyde treatment. This protocol allows the preparation of intact inactivated SARS-CoV-2 particles for safe use of samples out of level 3 laboratory, as revealed by combining AFM and plaque assays, to accelerate researches against the COVID-19 pandemic. Overall, we illustrate how adapted BSL3-atomic force microscopy is a remarkable toolbox for rapid and direct virus identification and characterization.

Download Full-text

Brittle Behaviour in Aspirin Crystals: Evidence of Spalling Fracture

10.26434/chemrxiv.13238708.v1 ◽

2020 ◽

Author(s):

Benjamin P. A. Gabriele ◽

Craig J. Williams ◽

Douglas Stauffer ◽

Brian Derby ◽

Aurora J. Cruz-Cabeza

Keyword(s):

Surface Roughness ◽

Atomic Force Microscopy ◽

Single Crystals ◽

Spalling Fracture ◽

Force Microscopy ◽

Atomic Force ◽

Afm Imaging

<div> <div> <div> <p>Single crystals of aspirin form I were cleaved and indented on their dominant face. Upon inspection, it was possible to observe strongly anisotropic shallow lateral cracks due to the extreme low surface roughness after cleavage. Atomic Force Microscopy (AFM) imaging showed spalling fractures nucleating from the indent corners, forming terraces with a height of one or two interplanar spacings d100. The formation of such spalling fractures in aspirin was rationalised using basic calculations of attachment energies, showing how (100) layers are poorly bonded when compared to their relatively higher intralayer bonding. An attempt at explaining the preferential propagation of these fractures along the [010] direction is discussed. </p> </div> </div> </div>

Download Full-text

Brittle Behaviour in Aspirin Crystals: Evidence of Spalling Fracture

10.26434/chemrxiv.13238708 ◽

2020 ◽

Author(s):

Benjamin P. A. Gabriele ◽

Craig J. Williams ◽

Douglas Stauffer ◽

Brian Derby ◽

Aurora J. Cruz-Cabeza

Keyword(s):

Surface Roughness ◽

Atomic Force Microscopy ◽

Single Crystals ◽

Spalling Fracture ◽

Force Microscopy ◽

Atomic Force ◽

Afm Imaging

Download Full-text

Atomic force microscopy (AFM) imaging suggests that stromal interaction molecule 1 (STIM1) binds to Orai1 with sixfold symmetry

FEBS Letters ◽

10.1016/j.febslet.2014.06.054 ◽

2014 ◽

Vol 588 (17) ◽

pp. 2874-2880 ◽

Cited By ~ 10

Author(s):

Dilshan Balasuriya ◽

Shyam Srivats ◽

Ruth D. Murrell-Lagnado ◽

J. Michael Edwardson

Keyword(s):

Atomic Force Microscopy ◽

Force Microscopy ◽

Stromal Interaction Molecule 1 ◽

Atomic Force ◽

Afm Imaging ◽

Sixfold Symmetry

Download Full-text

Imaging of Xenopus laevis Oocyte Plasma Membrane in Physiological-Like Conditions by Atomic Force Microscopy

Microscopy and Microanalysis ◽

10.1017/s1431927613001682 ◽

2013 ◽

Vol 19 (5) ◽

pp. 1358-1363 ◽

Cited By ~ 3

Author(s):

Massimo Santacroce ◽

Federica Daniele ◽

Andrea Cremona ◽

Diletta Scaccabarozzi ◽

Michela Castagna ◽

...

Keyword(s):

Atomic Force Microscopy ◽

Plasma Membrane ◽

High Resolution ◽

Membrane Proteins ◽

Xenopus Laevis ◽

Functional Study ◽

Xenopus Laevis Oocyte ◽

Force Microscopy ◽

Atomic Force ◽

Afm Imaging

AbstractXenopus laevis oocytes are an interesting model for the study of many developmental mechanisms because of their dimensions and the ease with which they can be manipulated. In addition, they are widely employed systems for the expression and functional study of heterologous proteins, which can be expressed with high efficiency on their plasma membrane. Here we applied atomic force microscopy (AFM) to the study of the plasma membrane of X. laevis oocytes. In particular, we developed and optimized a new sample preparation protocol, based on the purification of plasma membranes by ultracentrifugation on a sucrose gradient, to perform a high-resolution AFM imaging of X. laevis oocyte plasma membrane in physiological-like conditions. Reproducible AFM topographs allowed visualization and dimensional characterization of membrane patches, whose height corresponds to a single lipid bilayer, as well as the presence of nanometer structures embedded in the plasma membrane and identified as native membrane proteins. The described method appears to be an applicable tool for performing high-resolution AFM imaging of X. laevis oocyte plasma membrane in a physiological-like environment, thus opening promising perspectives for studying in situ cloned membrane proteins of relevant biomedical/pharmacological interest expressed in this biological system.

Download Full-text