High-throughput spatial proteomics in pulmonary research with FLASH multiplex imaging

Project Summary Addressing the complexities of heart, lung, and blood disorders at the cellular and molecular level requires a spatial omics technology capable of rapidly and reliably mapping the distribution of specific cell markers within tissue at the subcellular scale. While spatial proteomics technologies have revolutionized in situ molecular profiling of tissues, limitations in resolution, throughput, sample degradation, and cost have hindered their widespread impact. This is the focus of our project, which proposes to develop FLASH-PAINT, a disruptive proteomics technology, into AFLEX, an automated and multiplexed imaging solution specifically tailored for FFPE tissue samples. Our proposed technology promises to provide scientists with a simple method to perform exceptionally rapid 32-plex protein imaging on their tissue samples using a microfluidics chamber designed for easy integration with standard benchtop microscopes and necessitating minimal experimental intervention on their part. AFLEX is distinguished by its elimination of washing steps, thereby preventing sample degradation and enabling the rapid collection of highly multiplexed imaging data of large 1 cm2 tissue sections. Panluminate, Inc. is a leading company in the spatial proteomics space. Our proposed project aims to develop AFLEX and validate it in human FFPE tissue. We specifically propose to (1) develop a new concept and prototype for microfluidics tailored for FLASH samples that removes the need of traditional large-footprint microfluidics, (2) develop new FLASH probes designed for high signal-to-background imaging in FFPE tissue, and (3) test this system in a testbed application of fibrotic human lung tissue. Ultimately, the developments proposed here will enable panluminate to disseminate AFLEX imaging to every research and biotech lab around the world.