Imaging Mass Cytometry Laboratory

The laboratory is equipped with a Standard BioTools™ Hyperion XTi™ Imaging System, a multiplexed tissue imager that uses metal-tagged antibodies, instead of fluorophores, to simultaneously and reliably acquire 40-plus protein and RNA markers, without autofluorescence interference. 
Using the Hyperion XTi™ Imaging System, the IMC Laboratory performs spatial proteomics analyses of formalin-fixed, paraffin-embedded tissue samples; tissue microarrays; fresh-frozen tissues; and smeared cells on glass slides. We can stain specimens with metal-labeled antibodies and obtain high-dimensional images of up to 40 protein markers simultaneously, allowing researchers to interrogate the protein markers’ expression at subcellular resolution.

Working with researchers, we develop metal-labeled antibody panels then validate them with positive and negative control tissue slides/tissue microarrays. Once an antibody panel is finalized, we use 4–5µm sections of fresh-frozen or formalin-fixed, paraffin-embedded tissue, or smeared cells, then stain them with up to 40 different metal-labeled antibodies. The Regions of Interest (ROI) are ablated with the Hyperion XTi™ laser ablation unit then the content is ionized and transferred to a time-of-flight mass spectrometer, which resolves it based on its atomic mass.

After overlaying the single isotope signals for each ROI, we generate high-dimensional images that provide spatial and subcellular resolution of the tissue samples and the raw data from the instrument is shared with the investigator’s laboratory. If requested, we provide data analysis using project-dependent cell segmentation coupled with the Bodenmiller Lab pipeline for downstream analysis.