Design and characterization of a compact dual channel virus counter
CL Stoffel, RF Kathy, KL Rowlen - Cytometry Part A, 2005 - Wiley Online Library
CL Stoffel, RF Kathy, KL Rowlen
Cytometry Part A, 2005•Wiley Online LibraryBackground Although there is a growing need in the field of biotechnology to rapidly and
accurately quantify viruses, time‐consuming techniques such as the plaque titer method
remain the “gold standard.” Flow cytometric methods for virus quantification offer the
advantages of rapid analysis and statistical treatment. The technique presented in this work
represents the first demonstration of a flow cytometric determination of a viral count that is
directly related to the count obtained by plaque titer. Methods A flow cytometric instrument …
accurately quantify viruses, time‐consuming techniques such as the plaque titer method
remain the “gold standard.” Flow cytometric methods for virus quantification offer the
advantages of rapid analysis and statistical treatment. The technique presented in this work
represents the first demonstration of a flow cytometric determination of a viral count that is
directly related to the count obtained by plaque titer. Methods A flow cytometric instrument …
Background
Although there is a growing need in the field of biotechnology to rapidly and accurately quantify viruses, time‐consuming techniques such as the plaque titer method remain the “gold standard.” Flow cytometric methods for virus quantification offer the advantages of rapid analysis and statistical treatment. The technique presented in this work represents the first demonstration of a flow cytometric determination of a viral count that is directly related to the count obtained by plaque titer.
Methods
A flow cytometric instrument for rapid quantification of virus particles was designed, constructed, and thoroughly characterized. A two‐color method, which involved staining the viral genome and the protein coat for baculoviruses, was developed in addition to an algorithm to identify simultaneous events on the DNA and protein channels.
Results
The instrument was fully characterized, which included analysis of the data acquisition rate, sampling time, flow rate, detection efficiency, linear dynamic range, channel cross‐talk, and the limit of detection. Baculovirus samples were analyzed and the results were compared with concentrations obtained by a one‐channel flow cytometer and plaque assay.
Conclusions
The dual channel virus counter yields a representative value for the concentration of active viruses in an unpurified sample when compared with plaque assay and a one‐channel flow cytometer. The technique is rapid (within minutes), requires only minimal sample preparation and minimum sample size (∼100 μl). © 2005 Wiley‐Liss, Inc.
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