Abstract
Determination of virus abundance using epifluorescence microscopy is a rapid and accurate method. The protocol requires the concentration of virus particles by collection on a filter. The nucleic acid in the virus particles is then stained with a fluorescent stain and the sample viewed with an epifluorescence microscope. The method was originally developed to determine the abundance of virus particles in water samples, however the protocol has been adapted for cultures and sediment samples. Although the method provides total counts of all virus-sized particles, regardless of infectivity, the method can be used for rapidly screening samples for further study.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Ferris, Matthew M., Stoffel, Carrie L., Maurer, Thain T., and Rowlen, Kathy L. (2002). Quantitative intercomparison of transmission electron microscopy, flow cytometry, and epifluorescence microscopy for nanometric particle analysis. Anal. Biochem. 304, 249–256.
Børsheim, K.Y., Bratbak, G., and Heldal, M. (1990). Enumeration and biomass estimation of planktonic bacteria and viruses by transmission electron microscopy. Appl. Environ. Microb. 56, 352–356.
Proctor, L.M. and Fuhrman, J.A., (1990). Viral mortality of marine bacteria and cyanobacteria. Nature. 343, 60–62.
Hara, S., Terauchi, K., and Koike, I., (1991). Abundance of viruses in marine waters – Assessment by epifluorescence and transmission electron microscopy. Appl. Environ. Microb. 57, 2731–2734.
Porter, K.G. and Feig, Y.S., (1980). The use of DAPI for identifying and counting aquatic microflora. Limnol. Oceanogr. 25, 943–948.
Hennes, K.P. and Suttle, C.A., (1995). Direct counts of viruses in natural waters and laboratory cultures by epifluorescence microscopy. Limnol. Oceanogr. 40, 1050–1055.
Noble, R.T. and Fuhrman, J.A., (1998). Use of SYBR Green I for rapid epifluorescence counts of marine viruses and bacteria. Aquat. Microb. Ecol. 14, 113–118.
Chen, F., Lu, J.R., Binder, B.J., Liu, Y.C., and Hodson, R.E., (2001). Application of digital image analysis and flow cytometry to enumerate marine viruses stained with SYBR Gold. Appl. Environ. Microb. 67, 539–545.
Bettarel, Y., Sime-Ngando, T., Amblard, C., and Laveran, H., (2000). A comparison of methods for counting viruses in aquatic systems. Appl. Environ. Microb. 66, 2283–2289.
Wen, K., Ortmann, A.C., and Suttle, C.A., (2004). Accurate estimation of viral abundance by epifluorescence microscopy. Appl. Environ. Microb. 70, 3862–3867.
Danovaro, R., Dell’anno, A., Trucco, A., Serresi, M., and Vanucci, S., (2001). Determination of virus abundance in marine sediments. Appl. Environ. Microb. 67, 1384–1387.
Xenopoulos, M.A. and Bird, D.F., (1997). Virus á la sauce Yo-Pro: Microwave-enhanced staining for counting viruses by epifluorescence microscopy. Limnol. Oceanogr. 42, 1648–1650.
Suttle, C.A., (1993). Enumeration and isolation of viruses, in Handbook of Methods in Aquatic Microbial Ecology (Kemp, P.F, Sherr, B.F., Sherr E.B., and Cole, J.J., eds.) Lewis Publishers, Boca Raton, FL, pp. 121–134.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2009 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Ortmann, A.C., Suttle, C.A. (2009). Determination of Virus Abundance by Epifluorescence Microscopy. In: Clokie, M.R., Kropinski, A.M. (eds) Bacteriophages. Methods in Molecular Biology™, vol 501. Humana Press. https://doi.org/10.1007/978-1-60327-164-6_10
Download citation
DOI: https://doi.org/10.1007/978-1-60327-164-6_10
Publisher Name: Humana Press
Print ISBN: 978-1-58829-682-5
Online ISBN: 978-1-60327-164-6
eBook Packages: Springer Protocols