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 …
Divulgazioni
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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Citazione di questo articolo
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).