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Virus Propagation and Cell-Based Colorimetric Quantification
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High throughput virus plaque quantitation using a flatbed scanner.

Kate Sullivan1, Johannes Kloess, Chen Qian

  • 1Confocal Imaging and Analysis Laboratory, National Institute for Medical Research, The Ridgeway, Mill Hill, London NW7 1AA, UK.

Journal of Virological Methods
|November 3, 2011
PubMed
Summary
This summary is machine-generated.

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A new imaging method using a flatbed scanner and software automates influenza virus plaque counting. This cost-effective technique offers accurate quantification of virus infectivity and drug susceptibility, improving on subjective visual inspection.

Area of Science:

  • Virology
  • Cell Biology
  • Bioimaging

Background:

  • Plaque assays are standard for measuring influenza virus infectivity.
  • Accurate, automated quantification of viral plaques and infected cells remains challenging.
  • Visual inspection is subjective, time-consuming, and lacks reproducibility.

Purpose of the Study:

  • To develop a simple, high-throughput imaging-based method for quantifying influenza virus plaque formation and infected cells.
  • To provide an automated alternative to manual plaque counting.

Main Methods:

  • Utilized a flatbed scanner and image processing software for plaque quantification.
  • Compared automated quantitation results with visual counting.
  • Evaluated method performance in titrating plaques, infected cells, and determining neutralization titers and IC50s.

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Main Results:

  • Achieved over 80% agreement with visual counting.
  • Demonstrated high accuracy in titrating heterogeneous populations of small plaques.
  • Showed insensitivity to plaque density in neutralization and drug susceptibility assays.

Conclusions:

  • The proposed imaging method is a cost-effective, accurate, and readily available alternative for influenza virus plaque assays.
  • This technique enhances throughput and reproducibility in virological studies.
  • It is particularly advantageous for analyzing viruses with heterogeneous plaque sizes.