Single-molecule capture, release, and dynamical manipulation via reversible electrokinetic confinement (RECON).

We present a nanofluidic device enabling single-molecule confinement through free-energy landscapes created by dynamic electrical gating of embedded nanoelectrodes. Unlike static geometric confinement, this system uses a parallel electrode configuration with nanoelectrodes placed in a dielectric layer. Localized electrokinetic fields at electrode wells form tunable attractive potential wells for bimolecular capture. By modulating the voltage bias waveform, the device allows precise control over confinement dynamics, enabling molecular capture, release, and exposure to periodic or stochastic confinement regimes. This flexibility facilitates the study of biomolecular behavior under dynamically adjustable conditions, including controlled confinement fluctuations. The device can manipulate diverse analytes such as double-stranded DNA, liposomes, and DNA nanotubes and facilitates introducing molecules into confined environments intact from bulk while providing enhanced tunability. With the ability to implement tailored confinement profiles, this platform represents a versatile tool for probing molecular confinement and behavior in complex, dynamically varying environments.