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Presented by: Diana Fahrer, Associate Scientist III
Translational Pathology Laboratory at Biogen Idec
The “holy grail” of quantitative image analysis is to use intensity measurements of cells stained by immunohistochemistry (IHC) to accurately represent the amount of antigen bound by the primary antibody. Ideally, confidence in such an assay would be defined by using the same rigorous qualification standards as other ligand binding assays and would use chromogenic staining because of the contextural information brightfield staining provides. However, little is written about qualifying brightfield IHC assays and for the most part, intensity quantitation by IHC has relied on fluorophores due to the larger dynamic range they offer. This study represents an initial attempt to qualify a chromogenic IHC assay and to determine what situations, if any, chromogenic staining can be used to quantify intensity.