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Electronic Journal of Biotechnology
Universidad Católica de Valparaíso
ISSN: 0717-3458
Vol. 14, No. 1, 2011
Bioline Code: ej11011
Full paper language: English
Document type: Note
Document available free of charge

Electronic Journal of Biotechnology, Vol. 14, No. 1, 2011

 en Infectious pancreatic necrosis virus (IPNV) enumeration through epifluorescence microscopy: technical aspects
Pavlov, Maria Soledad; Eissler, Yoanna; Espinoza, Juan Carlos; Garland, David E. & Kuznar, Juan

Abstract

A method for counting Infectious pancreatic necrosis virus (IPNV) through epifluorescence microscopy was analyzed in detail. Image processing and statistic considerations are included. The particle size of viruses was compared in different experimental conditions such as the staining of the virus with SYBR-Green I or with antibodies for specific fluorescence labeling of viral proteins. The type of surface used as mounting support was assayed as well. The results indicated that the most suitable method involves the mounting of the viral-containing suspension on a membrane filter followed by the staining with a monoclonal antibody specific for a viral protein combined with a FITC (fluorescein isothiocyanate)-conjugated secondary antibody.

Keywords
digital image analysis, fluorescence microscope methods, immunofluorescence, Infectious pancreatic necrosis virus (IPNV), SYBR-Green I.

 
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