Current Methods in CODEX High-Multiplex Microscopy to Describe Spatial Relationships in the Tissue Microenvironment

Area of Science:

• Biomedical Engineering
• Molecular Biology
• Pathology

Background:

• Studying disease mechanisms at the cellular and tissue levels requires high-resolution imaging techniques.
• Existing methods may lack the multiplexing capability to analyze complex cellular interactions within tissues.

Purpose of the Study:

• To introduce and detail the CO-Detection by indEXing (CODEX) method for high-multiplex tissue imaging.
• To enable detailed spatial analysis of cell types, neighborhoods, and functional states within tissues.

Main Methods:

• Antibodies are conjugated to unique oligonucleotides.
• Conjugates are optimized for formalin-fixed paraffin-embedded (FFPE) or fresh-frozen (FF) tissues.
• CODEX involves a multicycle reaction with fluorescently labeled oligonucleotides for image capture, followed by image processing for spatial analysis.

Main Results:

• CODEX allows for high-multiplex imaging, analyzing up to 100 unique targets simultaneously.
• The method facilitates the identification of cell types, cellular neighborhoods, and their spatial relationships.
• Image processing enables cell type and neighborhood mapping for spatial analysis.

Conclusions:

• CODEX significantly advances basic antibody research and translational pathology.
• This high-resolution spatial analysis method refines clinical treatments by providing bioinformatic support for disease diagnosis, particularly in cancer.
• The technique offers a powerful tool for understanding complex biological systems at the microscopic level.