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Digital fluorescence imaging of fusion of influenza virus with erythrocytes
Author(s) -
Georgiou George N.,
Morrison Ian E.G.,
Cherry Richard J.
Publication year - 1989
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(89)80782-3
Subject(s) - haemolysis , fluorescence recovery after photobleaching , virus , fusion , biophysics , lipid bilayer fusion , fluorescence , microscopy , fluorescence microscope , photobleaching , chemistry , cell fusion , red blood cell , fluorescence lifetime imaging microscopy , live cell imaging , virology , membrane , cell , biology , optics , physics , biochemistry , philosophy , linguistics , immunology
Fusion of influenza virus with human erythrocytes at pH 5.2 was followed by fluorescence microscopy using a cooled slow‐scan CCD camera. The high sensitivity of the CCD permits repetitive digital imaging of the same cells with minimal photobleaching. The experimental conditions were such that only a small number of virus particles were adsorbed per cell. Quantitative analysis of the data indicated that for most cells only a single fusion event took place. This was, however, sufficient to cause haemolysis within 30 min at 20–22°C for about 60% of cells. There was a highly variable time lag between fusion and haemolysis. The lateral diffusion coefficient of virus particles on the cell surface when bound at pH 7.4 was < 2 × 10 −13 cm 2 ·s −1 . The technique should be of value for more detailed studies of the dynamics of viral and other membrane fusion events.