Calabrase, WilliamBishop, Logan D.C.Dutta, ChayanMisiura, AnastasiiaLandes, Christy F.Kisley, Lydia2020-12-162020-12-162020Calabrase, William, Bishop, Logan D.C., Dutta, Chayan, et al.. "Transforming Separation Science with Single-Molecule Methods." <i>Analytical Chemistry,</i> 92, no. 20 (2020) American Chemical Society: 13622-13629. https://doi.org/10.1021/acs.analchem.0c02572.https://hdl.handle.net/1911/109732Empirical optimization of the multiscale parameters underlying chromatographic and membrane separations leads to enormous resource waste and production costs. A bottom-up approach to understand the physical phenomena underlying challenges in separations is possible with single-molecule observations of solute–stationary phase interactions. We outline single-molecule fluorescence techniques that can identify key interactions under ambient conditions. Next, we describe how studying increasingly complex samples heightens the relevance of single-molecule results to industrial applications. Finally, we illustrate how separation methods that have not been studied at the single-molecule scale can be advanced, using chiral chromatography as an example case. We hope new research directions based on a molecular approach to separations will emerge based on the ideas, technologies, and open scientific questions presented in this Perspective.engThis is an open access article published under an ACS AuthorChoice License, which permits copying and redistribution of the article or any adaptations for non-commercial purposes.Journal articleacs-analchem-0c02572https://doi.org/10.1021/acs.analchem.0c02572