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We present a method to probe molecular and nanoparticle diffusion withinthin, solvated polymer coatings. The device exploits the confinement withwell-defined geometry that forms at the interface between a planar and ahemi-spherical surface (of which at least one is coated with polymers) in closecontact, and uses this confinement to analyse diffusion processes withoutinterference of exchange with and diffusion in the bulk solution. With thismethod, which we call plane-sphere confinement microscopy (PSCM), informationregarding the partitioning of molecules between the polymer coating and thebulk liquid is also obtained. Thanks to the shape of the confined geometry,diffusion and partitioning can be mapped as a function of compression andconcentration of the coating in a single experiment. The method is versatileand can be integrated with conventional optical microscopes, and thus shouldfind widespread use in the many application areas exploiting functional polymercoatings. We demonstrate the use of PSCM using brushes of natively unfoldednucleoporin domains rich in phenylalanine-glycine repeats (FG domains). Ameshwork of FG domains is known to be responsible for the selective transportof nuclear transport receptors (NTR) and their macromolecular cargos across thenuclear envelope that separates the cytosol and the nucleus of living cells. Wefind that the selectivity of NTR uptake by FG domain films depends sensitivelyon FG domain concentration, and that the interaction of NTRs with FG domainsobstructs NTR movement only moderately. These observations contribute importantinformation to better understand the mechanisms of selective NTR transport.

A method to quantify molecular diffusion within thin solvated polymer films: A case study on films of natively unfolded nucleoporins