Capsule-Based Negative Staining of Virus Samples on TEM Grids: A Heavy Metal Staining Technique to Visualize the Ultrastructure of Enveloped Viruses

Published: April 30, 2023

Abstract

Source: Blancett, C. D., et al. Utilization of Capsules for Negative Staining of Viral Samples within Biocontainment. J. Vis. Exp. (2017)

In this video, we demonstrate the negative staining of enveloped virus samples using a processing capsule within the biocontainment environment. The negatively stained viruses can be visualized using transmission electron microscopy to investigate viral structure and morphology with nanometer resolution.

Protocol

1. The Capsule Method for Inactivation in Biocontainment with 2% Glutaraldehyde, Followed by Negative Staining in a BSL-2 Laboratory Virus inactivation procedure.  Inside the biocontainment BSC, mix the virus suspension well with the same volume of 4% glutaraldehyde to achieve a final concentration of 2% glutaraldehyde. Caution: Glutaraldehyde is a hazardous chemical and requires appropriate protection. Glutaraldehyde can be used for …

Divulgations

The authors have nothing to disclose.

Materials

Formvar/carbon coated TEM grids  SPI  3420C-MB  200 mesh Cu Pk/100
mPrep/g capsules  EMS  85010-01  Box
mPrep/f couplers  EMS  85010-11  Standard 16/Pk
Glutaraldehdyde  EMS  16320 50% solution, EM grade
Uranyl Acetate  EMS  22400 Powder
Potassium phosphotungstic acid  EMS  19500 Powder
Filter paper  Whatman  1450-090  Size 50
Tranmission Electron Microscope  JEOL  JEM-1011  TEM

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Citer Cet Article
Capsule-Based Negative Staining of Virus Samples on TEM Grids: A Heavy Metal Staining Technique to Visualize the Ultrastructure of Enveloped Viruses. J. Vis. Exp. (Pending Publication), e21034, doi: (2023).

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