Allows Detection of 2–4 Markers Using 1° Antibodies from Any Species

High definition chromogenic multiplex IHC staining using UltraPlex technology allows precise visualization of tissue biomarkers. Our rapid staining technology and simple workflow using cocktails of antibodies and a single antigen retrieval step allows you to go from staining to analysis in a single day.

Routine multiplex immunohistochemistry detection has been limited to two chromogens using primary antibodies from different species detected using enzyme-labeled secondary anti-species antibodies followed by chromogen precipitation. Cell IDx’s UltraPlex hapten technology overcomes many of these limitations allowing detection of 2–4 markers using primary antibodies from any species employing a single antigen retrieval step on an autostainer.

Chromogenic Multiplex Staining of PD-L1/CD8/panCK

UltraPlex mxIHC staining of PD-L1/CD8/PanCK panel on tonsil. Note: PD-L1 co-localized with PanCK produces a dark green color.

Chromogenic Multiplex Staining of CD4/CD8/panCK

UltraPlex mxIHC staining of CD4/CD8/PanCK on colorectal cancer tissue .

Chromogenic Multiplex Staining of CD3/CD31/Ki-67

UltraPlex mxIHC staining of CD3/CD31/Ki-67 on tonsil.
Above images from an Aperio GT450 scanner.

To demonstrate that the UltraPlex IHC and UltraPlex IF technologies are detecting the same biomarkers, two serial tonsil slides were incubated with the same hapten-labeled primary cocktail and one slide was developed chromogenically and the other fluorescently using their respectively-labeled anti-hapten antibodies. The results are presented below using the PD-L1/CD8/GranzymeB panel.

Chromogenic Multiplex Staining of PD-L1/CD8/GranzymeB

mxIHC and mxIF images on serial section of tonsil with the PD-L1/CD8/GranzymeB three-plex as well as H&E staining of mxIF slide following removal of its coverslip.

The UltraPlex mxIHC protocol requires 4.5 hours for 3-marker staining, as illustrated below. The initial step in this mxIHC protocol is identical to Cell IDx’s multiplex immunofluorescence protocol in which a cocktail of hapten-labeled primary antibodies are incubated on the tissue. For chromogen detection, enzyme-labeled anti-hapten antibodies are employed instead of fluor-labeled anti-hapten antibodies.

UltraPlex Chromogenic mxIHC Workflow

Schematic presentation of UltraPlex mxIHC workflow to stain 3 chromogens on a single tissue using 1 antigen retrieval step.
  • For Research Use Only. Not for use in diagnostic procedures.

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