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Multiplexed Spatial Proteomics Services: Phenocycler aka CODEX

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Introduction

Highly multiplexed spatial proteomics makes it possible to visualize 10s of proteins and biomarkers, within multiple cell and tissue structures on a single tissue section, thus providing meaningful context. This enables powerful modeling of biological processes in context to and relationship with the multitude of proteins and biomarkers that define specific cell phenotypes and tissue architecture. One such technology, pioneered and developed in Dr. Nolan’s lab at Stanford and now licensed to Akoya Biosciences, is "CODEX" aka "PhenoCycler". This technology makes use of a DNA barcode technology comprised of unique, complimentary, oligonucleotide sequences. One unique oligo sequence is conjugated to a primary antibody (against a protein or other biomarker target); the complimentary oligo sequence is conjugated to a fluorescent reporter dye. DNA base pairing, between the conjugated complimentary oligonucleotides, then enables highly specific detection. Iterative cycles of imaging 3 different conjugated fluorophores (reports) at a time, plus a DNA dye, results in detection of 10s of biomarkers in a single tissue slice.

The CSIF offers a full array of highly multiplexed spatial proteomics services to Stanford researchers as well as to external researchers from academia and industry. These services include:

  • Custom panel development for mouse and human FFPE and FF tissue samples.
  • Custom antibody conjugation and validation.
  • Highly multiplexed Akoya or custom panel runs on PhenoCycler or CODEX instruments.
  • Data processing using Akoya’s processing software.
  • Data analysis using Multiplex Analysis Viewer (MAV) or VisioPharm software.

For more information contact lead scientist Yuanyuan Li, or director Jon Mulholland.

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Overview of CODEX iterative imaging cycles

More about CODEX/PhenoCycler on Akoya’s website

Contact information

Yuanyuan Li yuanyli@stanford.edu

Jon Mulholland jwm@stanford.edu

Literature (login required) and Useful Links

Useful papers and reviews:

  1. Spatial mapping of protein composition and tissue organization: a primer for multiplexed antibody-based imaging

Nolan lab Stanford:

  1. CODEX multiplexed tissue imaging with DNA-conjugated antibodies
  2. Deep Profiling of Mouse Splenic Architecture with CODEX Multiplexed Imaging.
  3. Coordinated Cellular Neighborhoods Orchestrate Antitumoral Immunity at the Colorectal Cancer Invasive Front.
  4. Strategies for Accurate Cell Type Identification in CODEX Multiplexed Imaging Data

CSIF supported publications:

  1. Desmoplastic stromal signatures predict patient outcomes in pancreatic ductal adenocarcinoma
  2. cPLA2 blockade attenuates S100A7-mediated breast tumorigenicity by inhibiting the immunosuppressive tumor microenvironment
  3. Multiplex imaging of human induced pluripotent stem cell-derived neurons with CO-Detection by indEXing (CODEX) technology
  4. Multiomic analysis reveals conservation of cancer-associated fibroblast phenotypes across species and tissue of origin

Akoya Inc.

  1. Phenocycler/CODEX inventoried and screened antibodies and panels
  2. CODEX Support