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Cell Counting and Image Cytometry

Cytomegalovirus Neutralization Assay Using Brightfield

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Antibody Neutralization
Antibody Binding Inhibition
Antibody Neutralization
Cytopathic Effects
Viral Titre
Topic
Cytomegalovirus Neutralization Assay Using Brightfield
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

Cytomegalovirus neutralization assay using different sera measured by direct cell counting in brightfield in a 96-well plate

  1. The ARPE epithelial host cells are seeded in a 96-well microplate an incubated for 24 hours to allow for the cells to adhere
  2. The cells are then infected by cytomegalovirus and incubated with titrations of different sera for another 24 hours
  3. Finally, the cells are fixed and stained with primary anti-viral antibodies and Horseradish Peroxidose (HRP) secondary
  4. After staining, the cells are imaged and analyzed using the Celigo™ image cytometer to count the number of infected cells

Brightfield-based antibody neutralization assay using HRP

The example below shows the low number of infected cells with high serum concentration in brightfield using HRP

Cytomegalovirus neutralization assay using brightfield-1

Brightfield image of HRP labeled infected cells with high serum

The example image below shows the high number of infected cells with low serum concentration in brightfield using HRP

Cytomegalovirus neutralization assay using brightfield-2

Brightfield image of HRP labeled infected cells with low serum

The number of infected cells are counted and dose response curves are generated for 4 different sera, where the IC50 values are calculated. The Celigo is able to output the counted results directly to Microsoft Excel for analysis.

Cytomegalovirus neutralization assay using brightfield-3

Plate-view of imaged and analyzed wells, and serum dilution-dependent neutralization
plot in respect to the number of HRP-labeled infected cells

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

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