Single cell genomics has transformed biology by enabling unbiased molecular identification and characterization of the full panoply of cell types that make up a tissue. A 'marriage' between this molecular information, which includes an inventory of all the ligands and receptors expressed by each cell class, and the accompanying 3D spatial information about them will enable the comprehensive reconstruction of the active signaling centers within a given tissue. Here, by continuing a fruitful collaboration between a biologist and molecular biochemist, we propose to deliver a transformative new technology for 3D multi-dimensional molecular imaging in intact human tissue. A major strength of our approach is that we entirely bypass the intrinsic limitations of fluorescent imaging, namely endogenous autofluorescence, low signal to noise, and restriction to five or fewer data channels, by using lanthanide resonant energy transfer. By so doing, we will simultaneously overcome challenges in detection of individual signals and massively increase throughput. Furthermore, the technology we deliver will be both biologist 'friendly' and low cost, providing a widely available and practical platform that will be rapidly and broadly adopted by scientists worldwide.

PROJECT NARRATIVE Our research focuses on making it possible to efficiently analyze individual cells in intact human tissues. This capability will be instrumental for understanding the 'logic' of how tissues function and will provide a healthy 'blueprint' against which to compare organs in states of disease.