Abstract
Single-molecule fluorescence studies enable real-time observation of interactions between molecular machines using minuscule amounts of (potentially) precious samples. Experimental repetition is required for optimizations, to test different samples or conditions, and ultimately to perform technical and biological repeats for publication. Here we developed a multiplexed approach that allows us to significantly increase experimental sensitivity, reproducibility, and throughput of single-molecule fluorescence and fluorescence energy transfer imaging. By coupling this approach with nucleic acid barcoding, microfluidic handling systems, and advanced data processing, we aim to automate and simplify multiperspective single-molecule studies to accelerate the emerging field of dynamic structural biology.
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