دورية أكاديمية
Utilization of Capsules for Negative Staining of Viral Samples within Biocontainment.
| العنوان: | Utilization of Capsules for Negative Staining of Viral Samples within Biocontainment. |
|---|---|
| المؤلفون: | Blancett CD; Pathology Division, United States Army Medical Research Institute of Infectious Diseases (USAMRIID)., Monninger MK; Pathology Division, United States Army Medical Research Institute of Infectious Diseases (USAMRIID)., Nguessan CA; Pathology Division, United States Army Medical Research Institute of Infectious Diseases (USAMRIID)., Kuehl KA; Pathology Division, United States Army Medical Research Institute of Infectious Diseases (USAMRIID)., Rossi CA; Diagnostic Systems Division, United States Army Medical Research Institute of Infectious Diseases (USAMRIID)., Olschner SP; Diagnostic Systems Division, United States Army Medical Research Institute of Infectious Diseases (USAMRIID)., Williams PL; Diagnostic Systems Division, United States Army Medical Research Institute of Infectious Diseases (USAMRIID)., Goodman SL; Microscopy Innovations LLC., Sun MG; Pathology Division, United States Army Medical Research Institute of Infectious Diseases (USAMRIID); mei.g.sun.ctr@mail.mil. |
| المصدر: | Journal of visualized experiments : JoVE [J Vis Exp] 2017 Jul 19 (125). Date of Electronic Publication: 2017 Jul 19. |
| نوع المنشور: | Journal Article; Video-Audio Media |
| اللغة: | English |
| بيانات الدورية: | Publisher: MYJoVE Corporation Country of Publication: United States NLM ID: 101313252 Publication Model: Electronic Cited Medium: Internet ISSN: 1940-087X (Electronic) Linking ISSN: 1940087X NLM ISO Abbreviation: J Vis Exp Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: [Boston, Mass. : MYJoVE Corporation, 2006]- |
| مواضيع طبية MeSH: | Capsules/*therapeutic use , Microscopy, Electron, Transmission/*methods , Negative Staining/*methods, Specimen Handling |
| مستخلص: | Transmission electron microscopy (TEM) is used to observe the ultrastructure of viruses and other microbial pathogens with nanometer resolution. Most biological materials do not contain dense elements capable of scattering electrons to create an image; therefore, a negative stain, which places dense heavy metal salts around the sample, is required. In order to visualize viruses in suspension under the TEM they must be applied to small grids coated with a transparent surface only nanometers thick. Due to their small size and fragility, these grids are difficult to handle and easily moved by air currents. The thin surface is easily damaged, leaving the sample difficult or impossible to image. Infectious viruses must be handled in a biosafety cabinet (BSC) and some require a biocontainment laboratory environment. Staining viruses in biosafety levels (BSL)-3 and -4 is especially challenging because these environments are more turbulent and technicians are required to wear personal protective equipment (PPE), which decreases dexterity. In this study, we evaluated a new device to assist in negative staining viruses in biocontainment. The device is a capsule that works as a specialized pipette tip. Once grids are loaded into the capsule, the user simply aspirates reagents into the capsule to deliver the virus and stains to the encapsulated grid, thus eliminating user handling of grids. Although this technique was designed specifically for use in BSL-3 or -4 biocontainment, it can ease sample preparation in any lab environment by enabling easy negative staining of virus. This same method can also be applied to prepare negative stained TEM specimens of nanoparticles, macromolecules and similar specimens. |
| References: | Adv Virol. 2011;2011:403419. (PMID: 22312342) Viruses. 2015 Feb 20;7(3):857-72. (PMID: 25710889) Electron Microsc Rev. 1990;3(1):43-72. (PMID: 1715774) J Transl Med. 2015 Jul 15;13:228. (PMID: 26174690) Methods Mol Biol. 2007;369:107-42. (PMID: 17656749) New Microbiol. 2014 Oct;37(4):403-22. (PMID: 25387280) J Virol Methods. 2016 Dec;238:70-76. (PMID: 27751950) Lancet. 2000 May 13;355(9216):1713-7. (PMID: 10905259) Micron. 2006;37(2):91-106. (PMID: 16361103) Biochim Biophys Acta. 1959 Jul;34:103-10. (PMID: 13804200) Arch Virol. 1987;94(3-4):305-8. (PMID: 2437888) Clin Microbiol Rev. 2009 Oct;22(4):552-63. (PMID: 19822888) Bacteriol Rev. 1967 Dec;31(4):230-314. (PMID: 4865539) Nature. 1966 Oct 1;212(5057):84-5. (PMID: 4164924) |
| المشرفين على المادة: | 0 (Capsules) |
| تواريخ الأحداث: | Date Created: 20170727 Date Completed: 20180119 Latest Revision: 20190719 |
| رمز التحديث: | 20231215 |
| مُعرف محوري في PubMed: | PMC5612576 |
| DOI: | 10.3791/56122 |
| PMID: | 28745647 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 1940-087X |
|---|---|
| DOI: | 10.3791/56122 |