MSACL 2019 EU Abstract

Exploring Healthy and Tumor Tissue Microenvironment with Immuno-Oncology Markers Using Multiplexed Hyperion Imaging System

Dongxia Lin1, Jeremy Sarnecky1, Christina Loh2, Roberto Spada4, Mary-Kay Lippert3
1Reagents Operation, Fluidigm Corporation, South San Francisco, CA, USA; 2Reagents Development, Fluidigm Canada Inc., Markham, ON, Canada; 3Reagents Operation, Fluidigm Canada Inc., Markham, ON, Canada 4Fluidigm SARL, Avenue de l'Atlantique Les Ulis France

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Roberto Spada (Presenter) Fluidigm

Presenter Bio: Roberto has a PhD from the National Center for Biotechnology-CSIC in Madrid (Spain) specializing in immuno-oncology. After different positions at Merck and Milteny Biotec, Roberto joined Fluidigm in 2017 and is currently in charge of business development for the Mass Cytometry franchise (Helios and the Hyperion Imaging System) in all of EMEA.

Relevant Financial Disclosures (within past 24 months) Salary Fluidigm Conflict mitigation NOT REQUIRED.

INTRODUCTION:
To power immuno-oncology discovery, it is highly beneficial to explore healthy and tumor tissues with immuno-oncology markers using multiplexed analysis. The Hyperion™Imaging System uses novel technology for tissue imaging that enables multiplexed analysis of protein expression in a single tissue sample. This methodology uses tissue sections stained with a cocktail of antigen-specific antibodies conjugated to different metal isotopes. In this study, we demonstrate how to generate high-parameter images with immuno-oncology markers on the Hyperion Imaging System, facilitating Imaging Mass Cytometry™ (IMC™) applications.
OBJECTIVE:
In this study, we demonstrate how to generate high-parameter images with immuno-oncology markers on the Hyperion Imaging System.
METHODS:
The IMC workflow uses immunostained and dried samples of tissue sections (FFPE or cryosections) and/or cells attached to glass slides which are inserted into the ablation chamber of the Hyperion Imaging System which uses a 1um spot-size pulsed UV laser at 200Hz which ablates the tissue. Isotopes associated with each spot are transferred into the mass cytometer, where via ICP-MS TOF, these are detected and indexed against the source location, yielding an intensity map of the target proteins throughout the tissue.
The use of highly pure metal labels on antibodies provides a solution to the current challenges in multiplexed tissue imaging by separating signals based on differences in mass, which overcomes the limitations of fluorescence-based detection modalities.
RESULTS:
Using these optimized staining protocols, we generated images from various normal and tumor tissues to show a combination of 5 structural, 1 cancer, 3 nuclear, and 18 immuno-oncology markers simultaneously. Together with other tissue architectural details, different immune cell types were identified in both normal and tumor tissues. This image resolution allows the visualization of proteins in the membranous, cytoplasmic, and nuclear cell compartments.
CONCLUSION:
The Hyperion Imaging System makes it possible to deeply interrogate tissues and tumors at sub-cellular resolution while preserving the information in tissue architecture and cellular morphology, ideal for characterization of the tissue microenvironment across a breadth of disease research areas, including immune-oncology.