Robert William Figliozzi, Feng Chen, Albert Chi, Hsia Shao-Chung Victor.Using the inverse Poisson distribution to calculate multiplicity of infection and viral replication by a high-throughput fluorescent imaging system .VIROLOGICA SINICA, 2016, 31(2): 180-183.doi: 10.1007/s12250-015-3662-8
Citation:
Robert William Figliozzi, Feng Chen, Albert Chi, Hsia Shao-Chung Victor.
Using the inverse Poisson distribution to calculate multiplicity of infection and viral replication by a high-throughput fluorescent imaging system .VIROLOGICA SINICA, 2016, 31(2)
: 180-183.
http://dx.doi.org/10.1007/s12250-015-3662-8
Using the inverse Poisson distribution to calculate multiplicity of infection and viral replication by a high-throughput fluorescent imaging system
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Department of Pharmaceutical Sciences, School of Pharmacy and Health Professions, University of Maryland Eastern Shore, Princess Anne, Maryland 21853, USA
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Department of Natural Sciences, School of Agriculture and Natural Sciences, University of Maryland Eastern Shore, Princess Anne, Maryland 21853, USA
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Department of Mathematics and Computer Sciences, University of Maryland Eastern Shore, Princess Anne, Maryland 21853, USA
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Corresponding author:
Hsia Shao-Chung Victor, vhsia@umes.edu, ORCID: 0000-0001-7623-2842
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Published Date:
26 January 2016
Available online:
01 April 2016
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Abstract
Here, we introduce an additional method that we believe has merits with respect to reducing labor, decreasing wait times, improving objectivity, and decreasing long-term costs. This new method, called fluorescently labeled infected cell inoculum titration (FLICIT), utilizes high-throughput fluorescent microscopy instrumentation such as Cytation 3 from Biotek, and is therefore only applicable to transgenic viruses that cause host cells to express fluorescent proteins. For the purpose of developing this method, we used the recombinant virus HSV-1 strain 17-Syn+, which expresses green fluorescent protein (GFP) (Figure 1A) and exhibits the same replication pattern as the wild type counterpart (Foster et al., 1998). This new method relies on titrating the inoculum so that dilution ensures that the rate of infection is less than 50%, with cells infected by nearly one viral particle each; thus, each fluorescent signal can be attributed to a single viral particle.
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Proportional views
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