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Cell Counting and Image Cytometry

Zika Virus Microneutralization Assay

Section
Modern Virology Assays
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Sub Section
Antibody Neutralization
Antibody Binding Inhibition
Antibody Neutralization
Cytopathic Effects
Viral Titre
Topic
Zika Virus Microneutralization Assay
Automated Focus Reduction Neutralization Test Using GFP-expressing RSV
Automated Imaging and Detection of Cytolytic Plaques
Cytomegalovirus Neutralization Assay Using Fluorescence
Zika Virus Microneutralization Assay
Cytomegalovirus Neutralization Assay Using Brightfield

Zika virus microneutralization assay using different antibodies measured by direct cell counting in 96-well plate

  1. Raji cells and 6 different antibody-RVP complexes (GFP-expressing Zika virus) are infected and seeded in 96-well microplates following the protocol described above
  2. After 48 hours, the Celigo™ image cytometer was used to count the number of GFP positive infected cells in each well to generate a dose response curve for all tested antibody-RVP complexes
  3. Two sets of example images of antibody-RVP complexes treatment are shown below, where GFP positive infected cells can be observed at different antibody concentrations
Zika virus microneutralization assay-1

Antibody dose-dependent bright field and GFP overlay images of Zika infected cells

  1. After counting the number of infected cells and total cell count for each antibody-RVP complex and concentrations using the gating function in the Celigo, a dose-response plot of percentage infection is generated and IC50 values are calculated.
Zika virus microneutralization assay-2

GFP fluorescent intensity histogram for gating infected cell population

Directly outline and count non-infected and infected cell populations after gating

  1. The dose-dependent antibody neutralization IC50 results are directly compared to FACS flow cytometer technology and are highly comparable
Zika virus microneutralization assay-3
Zika virus microneutralization assay-4

Comparison of percent Zika virus infection corresponding to various antibody titrations between the Celigo and FACS flow cytometer

For research use only. Not for use in diagnostic procedures.

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