Other Publications (1)
Articles by Chrystal A. Nguessan in JoVE
Utilization of Capsules for Negative Staining of Viral Samples within Biocontainment Candace D. Blancett1, Mitchell K. Monninger1, Chrystal A. Nguessan1, Kathleen A. Kuehl1, Cynthia A. Rossi2, Scott P. Olschner2, Priscilla L. Williams2, Steven L. Goodman3, Mei G. Sun1 1Pathology Division, United States Army Medical Research Institute of Infectious Diseases (USAMRIID), 2Diagnostic Systems Division, United States Army Medical Research Institute of Infectious Diseases (USAMRIID), 3Microscopy Innovations LLC This protocol provides instruction for negative staining virus samples which can easily be used in BSL-2, -3, or -4 laboratories. It includes the use of an innovative processing capsule, which protects the transmission electron microscopy grid and provides the user easier handling in the more turbulent environments within biocontainment.
Other articles by Chrystal A. Nguessan on PubMed
Preparation of Viral Samples Within Biocontainment for Ultrastructural Analysis: Utilization of an Innovative Processing Capsule for Negative Staining Journal of Virological Methods. Dec, 2016 | Pubmed ID: 27751950 Transmission electron microscopy can be used to observe the ultrastructure of viruses and other microbial pathogens with nanometer resolution. In a transmission electron microscope (TEM), the image is created by passing an electron beam through a specimen with contrast generated by electron scattering from dense elements in the specimen. Viruses do not normally contain dense elements, so a negative stain that places dense heavy metal salts around the sample is added to create a dark border. To prepare a virus sample for a negative stain transmission electron microscopy, a virus suspension is applied to a TEM grid specimen support, which is a 3mm diameter fragile specimen screen coated with a few nanometers of plastic film. Then, deionized (dI) water rinses and a negative stain solution are applied to the grid. All infectious viruses must be handled in a biosafety cabinet (BSC) and many require a biocontainment laboratory environment. Staining viruses in biosafety levels (BSL) 3 and 4 is especially challenging because the support grids are small, fragile, and easily moved by air currents. In this study we evaluated a new device for negative staining viruses called mPrep/g capsule. It is a capsule that holds up to two TEM grids during all processing steps and for storage after staining is complete. This study reports that the mPrep/g capsule method is valid and effective to negative stain virus specimens, especially in high containment laboratory environments.