Abstract 3800: Next generation of spatial biology: High-throughput multiplexed Imaging Mass Cytometry™ with whole slide modes
Authors: Raza Qanber , Pfister Thomas D. , Chwee Jyh Yun , Lim Liang , Howell David , Parsotam Nikesh , King David , Loh Christina 
Published: March 2024

Abstract
Gaining spatial insights into the cellular composition of tumor tissue has tremendous potential to inform clinical and translational researchers about mechanisms behind spatial predictors of immunotherapy success, disease etiology and progression. Imaging Mass Cytometry™ (IMC™) is a high-plex spatial biology imaging technique that enables deep characterization of the diversity and complexity of the tumor microenvironment (TME). IMC supports detailed assessment of cell phenotype and function using 40-plus metal-tagged antibodies simultaneously on a single slide without issues associated with fluorescence-based spectral overlap, tissue autofluorescence or implementation of multiple washing and acquisition cycles.
Currently, IMC enables user-defined regions of interest (ROI) in tissues to evaluate cellular and structural composition. To enhance the IMC user experience, we developed two new whole slide imaging (WSI) modes which enable streamlined workflows using ultrafast preview mode (PM) and high-throughput tissue mode (TM). PM samples the entire tissue at predefined spacing to rapidly capture a low-resolution image of all expressed markers in the antibody panel. PM generates an image in minutes to enable informed ROI placements while leaving the stained tissue intact for higher-resolution imaging. PM and TM are designed so acquisitions can easily occur on the same slide without additional processing steps. TM rapidly acquires images of the whole tissue at lower resolution (7 μm pixel size) at a quality that can be used for quantitative analysis of the tissue spatial biology. Specifically designed for high-throughput applications, TM in combination with a newly available 40-slide loader for the Hyperion XTi™ Imaging System permits automated and continuous imaging of more than 40 large tissue samples (400 mm2 per tissue) per week.
We showcase the application of WSI modes using the newly developed Maxpar® Human Immuno-Oncology IMC Panel Kit. The 31-marker panel was combined with catalog antibodies to create a 40-plus-marker panel that expands the ability to conduct comprehensive high-plex tumor and immune cell profiling. Tumor tissue microarrays (TMA) and whole tumor tissue sections were stained with the expanded panel. Single-cell analysis of selected ROIs, on whole tumor sections and TMAs, guided by PM data successfully provided quantitative analyses of spatial biology at single-cell resolution. In parallel, TM on whole tumor sections followed by pixel-based analysis provided a spatially resolved quantitative assessment of specific tumor and immune components of the TME.
This work demonstrates the expanded capabilities of IMC and establishes it as a reliable high-plex spatial biology imaging platform with high-throughput imaging capabilities ideally suited for translational and clinical applications.
For Research Use Only. Not for use in diagnostic procedures.
Citation Format: Qanber Raza, Thomas D. Pfister, Jyh Yun Chwee, Liang Lim, David Howell, Nikesh Parsotam, David King, Christina Loh. Next generation of spatial biology: High-throughput multiplexed Imaging Mass Cytometry™ with whole slide modes [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3800.