Ultrahigh-throughput imaging of 3D tumor microenvironment at nanoscale

Project Summary This proposal aims to develop an integrated ultrahigh-throughput 3D volumetric super-resolution imaging system by combining a multiscale fluorescence mesoscope with Magnify Expansion Microscopy. Current imaging systems are largely limited to 2D tissue sections at single-cell resolution, which fails to capture the subcellular interactions critical in cancer research. Our innovative system will allow for rapid, large-area 3D imaging with exceptional detail, enabling the study of interactions between cancer and immune cells in situ. We will employ a novel specimen immobilization approach, advanced fluorescence staining techniques, and a fully automated workflow to enhance reproducibility and throughput. By using a genetically engineered mouse model of lung cancer, we will visualize subcellular changes in immune synapses and identify distinct spatial clustering during tumorigenesis and immune invasion. This project will advance the application of spatial biology in cancer research, offering new insights into immune evasion mechanisms and improving immunotherapy strategies. Our technology promises to revolutionize our understanding of cellular interactions in oncology, providing high- precision, large-scale 3D super-resolution imaging capabilities.