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Abstract Temperature is a fundamental yet underexploited control parameter in nanopore-based single-molecule sensing, critically influencing ionic transport, molecular dynamics, and biomolecular interactions. Despite its importance, precise temperature control remains challenging in nanopore experiments, as thermal regulation often introduces electrical noise, limits measurement bandwidth, or requires bulky and complex system integration. Here, we present a localized, chip-level Peltier-based thermal regulation platform integrated with glass nanopore sensing that overcomes these limitations. The system enables rapid and reversible temperature modulation from − 10 to 50 °C while maintaining exceptionally low electrical noise, with root-mean-square current fluctuations of only a few picoamperes, even at elevated temperatures. Combined with an effective bandwidth of 50 kHz and a sampling rate of 1 MHz, the platform supports high-fidelity nanopore recordings under tunable thermal conditions. We demonstrate temperature-dependent control of molecular translocation kinetics, signal amplitude, and temporal resolution using structured DNA carriers. This approach establishes a simple and scalable strategy for temperature-controlled, high-bandwidth single-molecule sensing, with broad implications for nucleic acid analysis, protein transport, and nanopore-based chemical and biophysical studies.
Li et al. (Fri,) studied this question.