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The Kennedy Institute of Rheumatology was the first in Europe to adopt the Cell DIVE™ technology in 2020 as part of a small research facility (SRF) that has steadily expanded in the past 2 years, with current capacity extending to over 70 members running more than 40 biomarkers across human, mouse and macaque tissues.

The Cell DIVE system has since resulted in a publication wherein Cell DIVE multiplex imaging was used to compare fibroblast markers across various human tissues. The Cell DIVE workflow involves iterative rounds of antibody-based staining, imaging and fluorophore bleaching. The DPOC facility has tested over 90 markers working on 12 different human tissue types, in addition to 18+ markers validated for mouse tissue samples. Expansion of the facility with both the Open-Top Light Sheet Microscope and Multiplexed Ion Beam Imaging microscope in May 2023 have provided key tools in bridging the gap between high resolution 2D imaging and rapid 3D whole mount imaging.

BACKGROUND

Multiplexed imaging of tissues involves the simultaneous visualisation of numerous (20-100) protein biomarkers at single-cell resolution, enabling unprecedented insights into the spatial architecture and cellular diversity within human tissues. Multiplexed imaging methods can involve fluorescence-based antibody staining or mass spectrometry approaches that allow cell phenotyping in a preserved spatial context to uncover previously hidden cell-cell interactions in disease processes. Access to both types of multiplex imaging methods with the Cell DIVE and MIBIscope, allow for the combination of high-throughput discovery and high-resolution analysis of multiple markers on single tissue slides. Advances in whole-slide imaging with multiplexing platforms like the Cell DIVE™, allow for automatic generation of pseudo-H&E images and detection of up to 60 biomarkers. The MIBI technology facilitates the simultaneous detection of 54 biomarkers using secondary ion mass spectrometry at extraordinary sensitivity and rapid acquisition rates. Expansion of DPOC to support GCP (Good Clinical Practice) imaging with the MIBIscope will add capability to support clinical trials and experimental medicine with GCP compliant multiplex imaging technology, permitting cell-based readouts of fibrotic human liver, skin, lung, heart, tumour biopsies and samples with large amounts of adipose tissue. In collaboration with the Multiplexing Platforms at Oxford (OMIG) we aim to improve upon single-cell segmentation and subsequent tissue classification. The DPOC facility is at the forefront of quantifying spatial relationships at subcellular resolutions, complementing the wealth of single-cell RNAseq data and increasing the potential for understanding cellular processes in human disease.

VISION

  • Merging multiplexed fluorescence imaging with single-cell genomics
    The integration of Cell DIVE with CITE-seq would combine protein and transcriptome measurements into a single-cell readout in a spatially resolved context. This will initially be performed in combination with fluorescence/chromogenic RNAscope®. Eventually to be applied on the MERSCOPE™ platform.
  • Increased throughput with additional instruments and robotics LED Photoirradiation workstation
    Gemini AutoStainer to automate tissue clearing, washing and bleaching for Cell DIVE as well as RNAscope protocols. Slideholder robotics to operate overnight and image large volume of slides (20+) for a single staining/bleach round. Analysis workstation dedicated to analysing Cell DIVE data using deep-learning based pipelines (DeepCell). See our analysis pipeline.
  • Combine 2D and 3D imaging through light-sheet microscopy of whole tissue samples.