Abstract
<jats:p>Solid-state nanopores are established tools for single-molecule analysis, yet their sensing performance is often constrained by the limited temporal resolution of ionic current recordings. This is particularly challenging given the short residence times of proteins, nanostructured barcodes, and targets bound to molecular carriers. To overcome this limitation, we report a platform utilizing a controllable pressure gradient to generate a hydrodynamic fluid flow that opposes the electrokinetic forces acting on the analyte, thereby decoupling molecular transport speed from the sensing voltage. Here, we systematically investigate the dynamics of passage under these voltage-pressure conditions for diverse analytes, ranging from linear DNA and DNA origami nanostructures to proteins. Crucially, we show that polymer folding probability is reduced and the instantaneous velocity profile throughout the translocation process is uniformly attenuated. Furthermore, the effective slowing down of low molecular weight proteins improves the detection rate and yields rich event signatures required for accurate fingerprinting. This platform also provides novel insights into the origin of enhanced current blockages and enables an independent measurement of the effective charge of DNA under our operating conditions. Ultimately, this technique transforms solid-state nanopores into highly tunable devices, unveiling complex protein signatures and barcoded DNA substructures often obscured by translocation speed.</jats:p>