Neuropathic pain is linked to ongoing activity of peripheral nerve fibers. Although we have some ideas on which components may contribute to this phenomenon, we are still in the dark on its underlying mechanism in human nociceptors and sensory neurons. New, innovative approaches are needed to improve our capability to investigate nociceptor’s spontaneous activity. In this symposium we aim to discuss cutting-edge tools that are providing new mechanistic information on the origin of spontaneous activity of peripheral nerve fibers at the cellular and molecular level. We will present new multi-electrode-array (MEA) recording tools (Aguiar) for assessing neuronal excitability in cell cultures. These tools can be used to their full translational potential by using induced pluripotent stem cell (iPSC) derived nociceptors, for which we present differentiation approaches (Röderer) and biophysical analysis of pain linked spontaneous activity and hyperexcitability (Eberhardt). In an attempt to recreate a nerve in a dish, we will present results of the combination of MEAs with microfluidics and their use with iPSC derived neurons. Experimental findings are integrated into an in silico simulation of a peripheral nerve ending (Tigerholm), linking wet lab results with observations during microneurography recordings in humans.