Off-the-Shelf Coding-Free Low-Cost Approach for Temperature Control of Nanopore Experiments

Le résumé fourni par la source

Temperature significantly influences the kinetics and thermodynamics of molecular transport within solid-state nanopores with an inextricable link to translocation velocities, signal amplitudes, and event capture rates. However, dynamic thermal regulation is rarely implemented in solid-state nanopore systems largely due to high system costs (for both hardware and software) and exposure to electromagnetic interference due to breaching of the Faraday cage system. To address these challenges, a fully off-the-shelf, coding-free, and cost-effective closed-loop fluidic system was developed, enabling precise temperature control (±0.2°C) across a broad operational range from ~5 °C to 90 °C. The system employs a localized fluidic circulation loop powered by an external pump, a thermoelectric cooler, a heating block, and a relay operated via temperature logic, thereby eliminating the need for specialized microcontroller programming and achieving stable thermal conditions within a custom-insulated, nested Faraday cage. Electrical noise analysis demonstrates that the system maintains adequate noise shielding: with 100 kHz lowpass filtering of the Axopatch 200B, the root-mean-square current (Irms) increases by only ~11% (from ~28 pA to ~32 pA). The platform's effectiveness is further validated through single-molecule translocation experiments of the protein apo-transferrin in a 4 M LiCl electrolyte. Developed with a total hardware investment of under $400, this robust, plug-and-play methodology broadens access to temperature-controlled nanopore experiments and supports biophysical characterization of macromolecular conformation and transport kinetics.

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