Electron Microscopy of Viral Fractions

LB-Core and Core samples were prepared as described (see Viral Fractionation) and prepared for EM by fixation in 1.5% glutaraldehyde / 2% EM-grade paraformaldehyde (TAAB) in 0.1M sodium cacodylate for 45 min at RT. Briefly, samples were treated with reduced osmium, tannic acid, dehydrated through an ethanol series and embedded in epon resin [96 (link)]. Sections were collected on formvar coated slot grids, stained with lead citrate and imaged using a transmission electron microscope (Tecnai T12, Thermo Fisher Scientific) equipped with a charge-coupled device camera (SIS Morada; Olympus).

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Bidgood S.R., Samolej J., Novy K., Collopy A., Albrecht D., Krause M., Burden J.J., Wollscheid B, & Mercer J. (2022). Poxviruses package viral redox proteins in lateral bodies and modulate the host oxidative response. PLoS Pathogens, 18(7), e1010614.

Publication 2022

Tecnai T12 SIS Morada

Cacodylate Citrate Dehydrated ethanol Device Epon Formvar Fractionation Glutaraldehyde Osmium Paraformaldehyde Resin Sodium Tannic acid Transmission electron microscope

Corresponding Organization : SIB Swiss Institute of Bioinformatics

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Variable analysis

independent variables

Preparation method (LB-Core and Core samples)

dependent variables

Viral structures observed using transmission electron microscopy

control variables

Fixation in 1.5% glutaraldehyde / 2% EM-grade paraformaldehyde in 0.1M sodium cacodylate for 45 min at RT
Treatment with reduced osmium, tannic acid
Dehydration through an ethanol series
Embedding in epon resin
Sectioning on formvar coated slot grids
Staining with lead citrate

controls

No positive or negative controls were explicitly mentioned in the provided information.

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