IBEX

IBEX is a novel software tool designed for high-content imaging of various cellular phenotypes and multicellular ecosystems found in metazoan tissues. It utilizes an iterative immunolabeling and chemical bleaching technique called ITERATIVE BLEACHING EXTENDS MULTIPLEXITY (IBEX). This approach allows for the simultaneous visualization of over 65 parameters in diverse tissues, including human organs relevant to international consortium efforts. IBEX is compatible with more than 250 commercially available antibodies and 16 distinct fluorophores, making it easy to adapt to different imaging platforms using standardized slides and nonproprietary imaging chambers. The entire process can be completed within two to five days by biologists with minimal lab experience at a relatively low cost. To facilitate widespread adoption, IBEX provides comprehensive guidelines on tissue processing, curated lists of validated antibodies, and tissue-specific panels for multiplex imaging. Additionally, the software includes automated tools for image alignment that can be utilized by users without prior programming experience through the use of open-