PhenoImager Assays and Reagents

Shifting the paradigm from visual IHC to quantitative IF with Opal

We’ve rebranded some of our products, learn more ›

CODEX is now PhenoCycler
Phenoptics is now Phenolmager

Image Multiple Tissue Biomarkers in Context

Akoya’s Opal™ Multiplex IHC kits make multiplex results accessible to anyone who works with standard immunohistochemistry. With Opal technology, you can select antibodies for simultaneous IHC detection based on performance rather than species. Opal kits are optimized for reliable Multispectral Imaging (MSI) for simultaneous measurement of three to eight IHC targets, plus a nuclear stain.

Opal enables you to:

  • Accomplish better phenotypic scoring and quantification by using signal amplification
  • Reduce overall sample and reagent consumption with increased plexing
  • Achieve excellent resolution with low background
  • Reach up to 4 logs dynamic range compared to 1 log using chromogenic assay detection techniques

Opal Kits & Reagents​

Opal Predesigned Panels for Automated PhenoImager Workflows

Opal Antibody Panel Kits provide pre-optimized, ready-to-use primary antibodies and detection reagents designed for use with automated staining and PhenoImager™ Fusion and PhenoImager™ HT (formerly Vectra Polaris) instruments and offer a validated end-to-end solution for unparalleled quantitative data in translational immuno-oncology research.

Key Benefits:
  • Easy-to-Use – Fully validated plug-and-play multiplex immunofluorescence staining protocol.
  • Flexible – Adjustable signal intensity for your sample’s unique biomarker expression levels.
  • Powerful – Unlock the full impact of whole slide multispectral workflows with Akoya’s kits when used with the PhenoImager Fusion and PhenoImager HT (formerly Vectra Polaris) and inForm analysis software.
Catalog #
Pan I/O Automation 6-Plex Panel - for PhenoImager Systems
Melanoma Automation 6-Plex Panel - for PhenoImager Systems

Opal Multiplex IHC Detection Kits​

Opal for multiplex immunofluorescence in formalin-fixed, paraffin-embedded (FFPE) tissue can be performed manually or via automation. Opal enable the development of multiplexed assays with balanced, quantitative signal for rare and abundant targets, imaged in a single, final scan with the Akoya portfolio of multispectral imagers, including the PhenoImager Fusion and PhenoImager HT.

These Opal detection kits allow user to perform multiplex staining using an automated staining platform. Automation provides the flexibility to support the dynamic demands of translational research

Catalog #
Opal 6-Plex Automation Detection Kit - for PhenoImager Systems
Opal 6-Plex Automation Detection Kit
Opal 3-Plex Automation Detection Kit
Opal 3-Plex anti-Rabbit Automation Kit

These Opal kits offer users a manual method that is similar to standard immunohistochemistry (IHC) making them accessible to many laboratories where standard IHC method development is performed.

Catalog #
Opal 6-Plex Manual Detection Kit - for PhenoImager Sytems
Opal 6-Plex Manual Detection Kit
Opal 3-Plex Manual Detection Kit
Solid Tumor Immunology 6-Plex Panel
Tumor Infiltrating Lymphocyte 6-Plex Panel
Immunology Discovery Panel Kit
Lymphocyte 3-Plex Panel
Opal 3-Plex anti-Rabbit Manual Kit

Standalone Opal Fluorophore reagent packs allow you to pick and choose your panel’s biomarker quantity and design.

Catalog #
Opal 480 Reagent Pack
Opal 520 Reagent Pack
Opal 540 Reagent Pack
Opal 570 Reagent Pack
Opal 620 Reagent Pack
Opal 650 Reagent Pack
Opal 690 Reagent Pack
Opal 780 Reagent Pack

These ready-to-use ancillary reagents, from diluents to secondary antibodies and blocking buffers, have been optimized specifically for Opal™ multiplex fluorescent immunohistochemistry protocols

Catalog #
1X Opal TSA Plus Amplification Diluent, 50mL
1X Opal TSA Plus Automation Amplification Diluent, 50mL
1X Opal TSA Antibody Diluent / Block, 100mL
10X AR9 Buffer, 250mL
10X AR6 Buffer, 250mL
Opal Anti-Ms + Rb HRP Antibody Kit, 50mL
Opal Anti-Rb HRP Antibody Kit, 50mL

TSA Reagents

Tyramide signal amplification (TSA® staining) typically provides 2-3 logs of sensitivity enhancement over standard fluorescent and hapten based detection methods. TSA staining increases sensitivity while maintaining resolution and dynamic range while allowing reduced consumption of primary antibodies.

Protocol optimization for TSA detection is critical for success with the technique, whether used as part of the PhenoImager multiplex imaging instruments or other detection system.

TSA plus cyanine 3 mouse brain
TSA® and TSA® Plus fluorophore and hapten labeled detection kits, for detection and amplification of signals in immunohistochemistry (IHC), immunofluorescence (IF) or in situ hybridization protocols.
Catalog #
TSA Biotin
50-150 slides
50-150 slides
50-150 slides
TSA Coumarin System
100–300 slides
TSA Cyanine 3
50-150 slides
TSA Cyanine 5
50-150 slides
TSA Fluorescein
50-150 slides
TSA Plus Cyanine 3.5
50-150 slides
TSA Plus Cyanine 5.5
50-150 slides
50-150 slides

Related Publications

Clinical workflow for quantifying PD-L1 in non-small cell lung cancer (NSCLC)
Professor Manuel Salto-Tellez, MD
Chair of Molecular Pathology, Queen’s University Belfast
Researchers at Queen’s University demonstrate that digital image analysis (DIA) of mIF using Opal 7-Color Automation IHC Kit is highly comparable to DIA on DAB IHC slides
Multi-institutional TSA-amplified Multiplexed Immunofluorescence Reproducibility Evaluation (MITRE) Study
Janis Taube, MD, MSc, et al.,
The Johns Hopkins Hospital, Akoya Biosciences, Providence Cancer Institute, MD Anderson Cancer Center, Bristol Myers Squibb, Yale University School of Medicine
A collaborative study across 6 institutions demonstrated that an Opal based six-plex mIF panel used for the characterization of the PD-1/PD-L1 axis across multiple sites was highly sensitive with high concordance and reproducibility observed across the sites.
Characterizing the TIME with TSA-Based Multiplex Immunofluorescence
Alexander D. Borowsky,
MDUniversity of California Davis
This UC Davis led study developed mIF protocols incorporating Opal tyramide signal amplification (TSA) for immune profiling the tumor microenvironment

Related Resources