Skip to main content
NCBI home page
Springer Nature - PMC COVID-19 Collection logoLink to Springer Nature - PMC COVID-19 Collection
. 1986;9(4):217–220. doi: 10.1007/BF01665758

A simple, rapid method for preparation of virus isolates from cell culture for electron microscopy

D E Skilling 1, J E Barlough 1, E S Berry 1,✉, A W Smith 1
PMCID: PMC7088600  PMID: 32214599

Summary

A simple procedure for the rapid preparation of virus isolates from cell culture for negative-contrast electron microscopy was devised. Using only conventional centrifugation steps (i.e. without ultracentrifugation), the procedure produced consistent, fine-quality preparations of a variety of virus types differing in size/shape and buoyant density.

Key words: virus isolates; cells, cultured; electron microscopy; centrifugation

References

  • 1.Almeida J. D. Practical aspects of diagnostic electron microscopy. Yale J. Biol. Med. 1980;53:5–18. [PMC free article] [PubMed] [Google Scholar]
  • 2.Almeida J. D. Rapid viral diagnosis by electron microscopy. Scand. J. Infect. Dis. [Suppl] 1982;36:117–120. [PubMed] [Google Scholar]
  • 3.Berry E. S., Shea T. B., Gabliks J. Two iridovirus isolates from Carassius auratus (L.) J. Fish Dis. 1983;6:501–510. [Google Scholar]
  • 4.Burroughs J. N., Doel. T. R., Smale C. J., et al. A model for vesicular exanthema virus, the prototype of the calicivirus group. J. Gen. Virol. 1978;40:161–174. doi: 10.1099/0022-1317-40-1-161. [DOI] [PubMed] [Google Scholar]
  • 5.Caul E. O., Ashley C. R., Egglestone S. I. An improved method for the routine identification of fecal viruses using ammonium sulfate precipitation. FEMS Microbiol. Lett. 1978;4:1–4. [Google Scholar]
  • 6.England J. J., Reed D. E. Negative contrast electron microscopic techniques for diagnosis of viruses of veterinary importance. Corne., Vet. 1980;70:125–136. [PubMed] [Google Scholar]
  • 7.Flewett T. H., Bryden A. S., Davies H. Diagnostie electron microscopy of faeces. I. The viral flora of the faeces as seen by electron microscopy. J. Clin. Pathol. 1974;27:603–614. doi: 10.1136/jcp.27.8.603. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 8.Hammond G. W., Hazelton P. R., Chuang I., et al. Improved detection of viruses by electron microscopy after direct ultracentrifuge preparation of specimens. J. Clin. Microbiol. 1981;11:210–221. doi: 10.1128/jcm.14.2.210-221.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 9.Horzinek M. C., Osterhaus A. D. M. E., Ellens D. J. Feline infectious peritonitis virus. Zentralbl. Veterinarmed. [B] 1977;24:398–405. doi: 10.1111/j.1439-0450.1977.tb01013.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 10.Lee F. K., Nahmias A. J., Stagno S. Rapid diagnosis of cytomegalovirus infection in infants by electron microscopy. N. Engl. J. Med. 1978;299:1266–1270. doi: 10.1056/NEJM197812072992302. [DOI] [PubMed] [Google Scholar]
  • 11.Madeley C. R. Comparison of the features of astroviruses and caliciviruses seen in samples of feces by electron microscopy. J. Infect. Dis. 1979;139:519–523. doi: 10.1093/infdis/139.5.519. [DOI] [PubMed] [Google Scholar]
  • 12.McFerran J. B., Clarke J. K., Curran W. L. The application of negative contrast electron microscopy to routine veterinary virus diagnosis. Res. Vet. Sci. 1971;12:253–257. [PubMed] [Google Scholar]
  • 13.Narang H. K., Codd A. A. A low-speed centrifugation technique for the preparation of grids for direct virus examination by electron microscopy. J. Clin. Pathol. 1979;32:304–305. doi: 10.1136/jcp.32.3.304. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 14.Narang H. K., Codd A. A. Enterovirus typing by immune electron microscopy using low-speed centrifugation. J. Clin. Pathol. 1980;33:191–194. doi: 10.1136/jcp.33.2.191. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 15.Polson A. New approaches to ultracentrifugation. In: Maramoroseh. K., Koprowski H., editors. Methods in virology, vol. V. New York: Academic Press; 1971. pp. 33–77. [Google Scholar]
  • 16.Portnoy B. L., Conklin R. H., Menn M., et al. Reliable identification of reovirus-like agent in diarrheal stools. J. Lab. Clin. Med. 1977;89:560–563. [PubMed] [Google Scholar]
  • 17.Rice S. J., Phillips A. D. Rapid preparation of faecal specimens for detection of viral particles by electron microscopy. Med. Lab. Sci. 1980;37:371–372. [PubMed] [Google Scholar]
  • 18.Rodgers F. G. Detection of rotavirus particles from patients with gastroenteritis. Lancet. 1980;1:1360–1361. doi: 10.1016/s0140-6736(80)91808-5. [DOI] [PubMed] [Google Scholar]
  • 19.Smith A. W., Skilling D. E., Ritchie A. E. Immunoelectron microscopic comparisons of caliciviruses. Am. J. Vet. Res. 1978;39:1531–1533. [PubMed] [Google Scholar]
  • 20.Stoddart C. A., Barlough J. E., Scott F. W. Experimental studies of a coronavirus and coronavirus-like agent in a barrier-maintained feline breeding colony. Arch. Virol. 1984;79:85–94. doi: 10.1007/BF01314306. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 21.Whitaker H. K., Alderson C. The use of negative contrast electron microscopy (NCEM) for diagnosis of viral infections in animals. Annu. Proc. Am. Assoc. Vet. Lab. Diagnost. 1980;23:321–350. [Google Scholar]
  • 22.Whitby H. J., Rodgers F. G. Detection of virus particles by electron microscopy with polyacrylamide hydrogel. J. Clin. Pathol. 1980;33:484–487. doi: 10.1136/jcp.33.5.484. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 23.Zissis G., Lambert J. P. DeKegel, D. Routine diagnosis of human rotaviruses in stools. J. Clin. Pathol. 1978;31:175–178. doi: 10.1136/jcp.31.2.175. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Tissue Culture Methods are provided here courtesy of Nature Publishing Group

RESOURCES