Transformative Technology Development
High-Throughput, Highly Multiplexed In Situ Proteomic Imaging of Human Tissues
This TTD utilizes a highly multiplexed signal amplification technique that can simultaneously amplify tens of distinct targets by tens- to hundreds-fold, thereby enabling highly multiplexed, high throughput, in situ imaging of proteins in human tissues. This in situ signal amplification method is based on a novel molecular mechanism recently published by this Harvard-based lab. In the new method, staining with multiple probes (DNA-barcoded primary antibodies) are performed simultaneously, and then all barcodes are simultaneously extended into long concatemers in situ. Finally, mapping of concatemers are sequentially performed through rapid exchange and imaging cycles, improving throughput by 10-fold while enabling detection of rare targets in tissues. Beyond proteins, the method is applicable to RNA and DNA (chromosome) targets. The method is integrated with commercially available, automated staining and imaging systems, and applied to image diverse human tissues via broad collaboration within and beyond the HuBMAP community.