A platform for high-bandwidth nanopore sensing with thermal control and low electrical noise

We present an instrument capable of performing high-bandwidth (1 MHz) solid-state nanopore measurements in a temperature-controlled environment ranging from ambient to 95 °C while maintaining low electrical noise. In previous systems, the ability to control the temperature of the analyte solution during nanopore sensing has come at the expense of significantly greater electrical noise. As a consequence, increased filtering requirements or, equivalently, reduced bandwidths ultimately decrease the utility of such instruments for biosensing applications. Here, we describe in detail the system we have developed that overcomes these difficulties. In particular, we are able to precisely control the temperature of the solution in which a nanopore sensor is immersed by using a closed-loop fluidics system. The ultra-low electrical conductivity heat transfer fluid is used to bring heat from outside of the Faraday cage to the nanopore sensor within the cage, resulting in minimal electrical noise during high-bandwidth measurements while maintaining localized temperature control. As proof-of-concept, we characterize silicon nitride nanopore stability over time at elevated temperatures using electrical measurements and present single-molecule data showing the impact of temperature on capture rate, dwell time, and blockage depth. This tool can unlock the ability to perform a wide range of temperature-sensitive biophysical experiments with solid-state nanopores.