We describe the incorporation of gated ion channels into probes for scanning ion conductance microscopy (SICM) as a robust platform for collecting spatial information at interfaces. Specifically, a dual-barrel pipet is used, where one barrel controls the pipet position and the second barrel houses voltage-gated transient receptor potential vanilloid 1 (TRPV1) channels excised in a sniffer-patch configuration. Spatially resolved sensing with TRPV1 channels is demonstrated by imaging a porous membrane where a transmembrane potential across the membrane generates local electric field gradients at pores that activate TRPV1 channels when the probe is in the vicinity of the pore. The scanning routine and automated signal analysis demonstrated provide a generalizable approach to employing gated ion channels as sensors for imaging applications.