Procedural requirements and recommendations for multiplex immunofluorescence tyramide signal amplification assays to support translational oncology studies

Edwin Roger Parra, Mei Jiang, Luisa Solis, Barbara Mino, Caddie Laberiano, Sharia Hernandez, Swati Gite, Anuj Verma, Michael Tetzlaff, Cara Haymaker, Auriole Tamegnon, Jaime Rodriguez-canales, Clifford Hoyd, Chantale Bernachez, Ignacio Wistuba

Research output: Contribution to journalReview articlepeer-review

48 Scopus citations

Abstract

In the development of a multiplex immunofluorescence (IF) platform and the optimization and validation of new multiplex IF panels using a tyramide signal amplification system, several technical requirements are important for high-quality staining, analysis, and results. The aim of this review is to discuss the basic requirements for performing multiplex IF tyramide signal amplification (TSA) in formalin-fixed, paraffin-embedded cancer tissues to support translational oncology research. Our laboratory has stained approximately 4000 formalin-fixed, paraffinembedded tumor samples using the multiplex IF TSA system for immune profiling of several labeled biomarkers in a single slide to elucidate cancer biology at a protein level and identify therapeutic targets and biomarkers. By analyzing several proteins in thousands of cells on a single slide, this technique provides a systems-level view of various processes in various tumor tissues. Although this technology shows high flexibility in cancer studies, it presents several challenges when applied to study different histology cancers. Our experience shows that adequate antibody validation, staining optimization, analysis strategies, and data generation are important steps for generating quality results. Tissue management, fixation procedures, storage, and cutting can also affect the results of the assay and must be standardized. Overall, this method is reliable for supporting translational research given a precise, step-by-step approach.

Original languageEnglish (US)
Article number255
JournalCancers
Volume12
Issue number2
DOIs
StatePublished - Feb 2020

Keywords

  • Image analysis
  • Multiplex immunofluorescence
  • Tumor immune profiling
  • Tyramide signal amplification

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

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