Highly-Multiplexed Fluorescence with DNA-Nanoruler Spectral Barcodes

Clemson University, Clemson SC

Investigators

Abstract

An award is made to Clemson University to enable a five to ten-fold increase in the number of different things that can be measured in a mixture simultaneously (multiplexing), using widely available, affordable, and reliable methods. This project will have broader impacts related to enhanced scientific access to highly-multiplexed experimental capabilities. It will provide training for the next generation of scientists by engaging undergraduates and high school students from a variety of disciplinary backgrounds and by mentoring doctoral students to carry out the work. Findings will be integrated with courses at Clemson University and with continued implementation of new student learning assessment methods. They will be available by open-source access of results, protocols, raw data, and analysis tools, and by public outreach through the Clemson University student chapters of scientific societies. Fluorescent reagents are a workhorse across the biological sciences. Conventional fluorescence multiplexing is limited to about four colors simultaneously, with recent advances reporting from 20 to 40 colors. This project will enable about 200 colors, which is a five-ten-fold increase. The approach is compatible with other current methods, which could expand multiplexing another five-ten-fold, to between 1,000 and 2,000 measurements. Increased measurement multiplexing usually enables breakthrough advances in biological understanding. The recent availability of compatible and affordable hardware will drive applications and user communities. Developmental and tissue biology questions could be probed with unprecedented depth by high-dimensional profiling. Overlaying high-dimensional data from this research in a spatial tissue context will synergize with ongoing efforts to build comprehensive maps of every single cell in the body. Microbial communities from skin, gut, soil, or ocean could be stratified, isolated, studied and combined with unprecedented precision. The generality of fluorescence combined with assay accessibility will enable wide adoption by user communities in multiple disciplines to answer novel biological and other scientific questions. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.