Imaging Mass Cytometry Laboratory

Overview

The Imaging Mass Cytometry Laboratory provides cancer researchers with high-dimensional imaging analysis of tumor tissue samples at subcellular resolution.

We work with researchers interested in studying the tumor microenvironment, tumor-stromal cells interactions, and the roles of different subpopulations of immune cells in tumor progression and suppression in animal models and cancer patients samples. 

The laboratory is equipped with a  Fluidigm Hyperion Imaging System that performs high-dimensional proteomics analysis of fresh-frozen tissues, tissue microarrays, smeared cells and formalin-fixed, paraffin-embedded tissue.  

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Fluidigm Hyperion Imaging System

With this technology, we use metal- labeled antibodies to stain the specimens on glass slides and obtain high-dimensional images of up to 40 protein markers simultaneously. This allows researchers to interrogate the protein markers expression in the nucleus and cytoplasm, as well as on the cell membrane. 

In addition to the tissue slides analysis, our laboratory has implemented the technology for multiplex proteomics characterization of rare circulating tumor cells isolated from cancer patients’ blood samples. This helps to better understand the role of CTCs in the metastatic spread of a disease and to study drug resistance and mutational switchover.  

We work with researchers to design customized metal-label antibody panels, validate the panels, and optimize the staining protocols. We also maintain a state-of-the-art data analysis pipeline for quantitative data analysis of high-dimensional, single-cell data generated by our laboratory.   

Innovations

We have developed and validated the following metal-labeled antibody panels that are available to researchers: 

  • Epithelial to mesenchymal transition markers 
  • Metabolic protein markers 
  • Tumor microenvironment  
  • Protein markers targeted to the specific cancer types 
  • Proliferation markers  
  • Cancer stem cell markers 

The Imaging Mass Cytometry Laboratory has established data analysis pipelines:  

  • Cell profiler- histoCAT-based data analysis pipeline from Bodenmiller laboratory  
  • Halo from Indica laboratory  

Focus

We work with human and murine fresh-frozen or formalin-fixed, paraffin-embedded tissue samples, smeared cells, rare circulating tumor cells, cell lines and patient-derived primary cultures. 

We maintain a repository of more than 50 validated metal-labeled antibodies targeted to the EMT, cancer stem cells, immune cells, and cell metabolism pathway. 

Our expert staff performs metal-label antibody staining, laser ablation of tissue samples and high-dimensional imaging analysis. Our services include: 

  • Metal-label antibody panel design customized for each research project. 
  • Assistance in procurement of commercially available metal-label antibodies. 
  • In-house metal-label antibody conjugation of other antibodies that are not commercially available. 
  • Validation of all the metal-labeled antibodies with known positive and negative control tissue samples or cell lines.

Technology

We use fresh-frozen or formalin-fixed, paraffin-embedded tissue sections of 4-5 µm size or smeared cells to stain with up to 40 different metal-labeled  antibodies. Different regions of interest on glass slides are then ablated with a Hyperion laser ablation unit using a 1 µm spot size. The content is ionized and transferred to the time-of-flight mass spectrometer where it is resolved based on the atomic mass.

After overlaying all the single isotope signals for every spot for all the laser shots, we generate high dimensional images—achieving spatial and subcellular resolution of tissue samples.