Rapid Multi-Parameter Tissue Imaging
While tissue imaging studies to date have been highly illuminating, the low number of simultaneous measurements has limited their power. Current systems measure up to five parameters using typical microscopes, and up to twelve using special instruments. This limitation is largely based on the number of fluorophores which can be simultaneously imaged. Approaches have been developed to overcome such limitations, but to date, they are primarily brute force methods which require multiple time-consuming stain/strip/wash cycles and can degrade the sample over time.
As such, deep proteomic profiling that capitalizes on knowledge of tissue organization in situ remains a critical unmet need. High parameter imaging is essential in deciphering the complexity of cellular populations with a dynamically changing composition such as a bone marrow or tumor microenvironments.
Our CODEX™ (CO-Detection by indEXing) technology enables this high parameter deep proteomic profiling, initially measuring 30-50 parameters in less than a day. The technology utilizes a “single stain, multiple images” approach.
The tissue is first labeled with the entire antibody panel in one step (~50 markers). Antibody binding events are then revealed and imaged in cycles by adding and removing cognate CODEX-tagged dyes.
This process is iterated such that the binding of 30-50 antibodies has been analyzed for all cells in the tissue section. Fluorescent signals from each round are computationally combined into a multi-channel image stack and can be subjected to image segmentation, quantification, and analysis.
This multidimensional data enables the user to identify novel cellular neighborhoods and delineate profoundly different tissue micro-architectures, providing a basis for a wide range of novel applications, particularly within immunology and oncology.
Noemi Kedei, M.D.
Collaborative Protein Technology Resource
OSTR, Office of the Director, CCR, NCI
37 Convent drive Room 1044
Bethesda, MD, 20892, USA