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Accurate virus quantitation using a Scanning Transmission Electron Microscopy (STEM) detector in a scanning electron
Candace D Blancett1, David P Fetterer2, Keith A Koistinen1
1Pathology Division, United States Army Medical Research Institute of Infectious Diseases (USAMRIID), 1425 Porter Street, Fort Detrick, MD, 21702, United States.
Journal of Virological Methods
|July 3, 2017
Summary
Accurate virus particle quantitation is crucial. A new Scanning Transmission Electron Microscopy - Virus Quantitation (STEM-VQ) method offers improved accuracy and reproducibility for counting viral particles.
Area of Science:
- Virology
- Microscopy
- Biotechnology
Background:
- Accurate quantitation of virus particles is essential but challenging.
- Traditional Electron Microscopy (EM) methods for virus quantitation have limitations in accuracy, reproducibility, and detection limits.
- There is a need for improved, reliable methods for determining viral particle counts.
Purpose of the Study:
- To develop and validate a novel, accurate, and reproducible method for virus particle quantitation.
- To introduce Scanning Transmission Electron Microscopy - Virus Quantitation (STEM-VQ) as an improved EM technique.
- To compare the performance of STEM-VQ against established methods like viral plaque assay and ViroCyt Virus Counter.
Main Methods:
- Development of the Scanning Transmission Electron Microscopy - Virus Quantitation (STEM-VQ) method.
- Simplified sample preparation protocols for electron microscopy.
- Utilizing a high-throughput STEM detector within a Scanning Electron Microscope (SEM) and commercial software.
- Quantitation of an alphavirus stock using STEM-VQ.
Main Results:
- Demonstration of STEM-VQ's accuracy and reproducibility in quantitating virus particles.
- Comparative analysis showing STEM-VQ's performance against viral plaque assay and ViroCyt Virus Counter.
- Validation of STEM-VQ as a reliable technique for viral particle counting.
Conclusions:
- STEM-VQ offers a significant improvement over traditional EM quantitation methods.
- The developed STEM-VQ method provides accurate and reproducible virus particle counts.
- STEM-VQ is a valuable tool for virology research requiring precise viral load determination.
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