期刊
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
卷 -, 期 78, 页码 -出版社
JOURNAL OF VISUALIZED EXPERIMENTS
DOI: 10.3791/50723
关键词
Virology; Issue 78; Infection; Immunology; Medicine; Molecular Biology; Cellular Biology; Microbiology; Genetics; Microscopy; Fluorescence; Neurobiology; Herpes virus; fluorescent protein; epifluorescent microscopy; neuronal culture; axon; virion; video microscopy; virus; live cell; imaging
资金
- US National Institutes of Health [R37 NS033506-16, R01 NS060699-03]
- American Cancer Society Postdoctoral Research Fellowship [PF-10-057-01-MPC]
Advances in live cell fluorescence microscopy techniques, as well as the construction of recombinant viral strains that express fluorescent fusion proteins have enabled real-time visualization of transport and spread of alphaherpes virus infection of neurons. The utility of novel fluorescent fusion proteins to viral membrane, tegument, and capsids, in conjunction with live cell imaging, identified viral particle assemblies undergoing transport within axons. Similar tools have been successfully employed for analyses of cell-cell spread of viral particles to quantify the number and diversity of virions transmitted between cells. Importantly, the techniques of live cell imaging of anterograde transport and spread produce a wealth of information including particle transport velocities, distributions of particles, and temporal analyses of protein localization. Alongside classical viral genetic techniques, these methodologies have provided critical insights into important mechanistic questions. In this article we describe in detail the imaging methods that were developed to answer basic questions of alphaherpes virus transport and spread.
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