A critical barrier to constructing molecular and cellular tumor atlases is the lack of spatialomics tools that can rapidly and cost-effectively profile multi-omics in large and thick tumor tissues with molecular resolution. Existing methods are either limited spectrally to 3-4 targets, as is the case of conventional fluorescence in situ hybridization and immunohistochemistry, or limited by complexity, error-prone image registration, poor spatial resolution, long assay time and high cost as with indirect sequential or sequence-based methods. This SBIR project aims to develop a new 3D spatial multi-omics technology employing integrated fluorescence hyperspectral/lifetime light sheet imaging system and combinatorial fluorescence spectral and lifetime encoded in situ labeling probes. If successful, this technology will be capable of direct, in situ profiling a large number of RNA and protein markers in large tumors or whole biopsies at high speed with molecular resolution with one round of staining and imaging. This technology will be developed using fully characterized xenografted colorectal cancer tumors and clinical melanoma formalin fixed paraffin embedded tissues as testbeds and benchmarked with gold standard methods. This disruptive technology can be quickly and broadly adopted to expedite building tumor atlases at a substantially lower cost and enable companion diagnostic-stratified precision healthcare.
