Introduction: Habituation to pain is a generally known phenomenon that involves a decrease in response after repeated painful stimulation 1. A variety of methods is available to measure short-term habituation. Nevertheless, the neural correlates are not well understood. Therefore, we developed a protocol integrating psychophysical and neural measures. Methods: Participants received three blocks of 25 brief painful electric stimuli while fMRI data was collected. After each stimulus, participants rated their pain on a visual analogue scale (VAS). The protocol was designed in a way to include ratings, allow for habituation and limit the influence of motor responses on the pain response. Analysis of habituation includes multilevel models for pain ratings and brain activity over blocks and on a trial-by-trial basis. Results: Preliminary results (n = 5) indicated a decrease in pain ratings, with large individual differences. In addition , we found brain activity in areas involved in pain processing in general (i.e., insula, cingulate cortex), which showed decreased activity over blocks of painful stimulation. Furthermore, in more refined analyses (trial-by-trial), we expect to find a linear decrease in activity in the anterior/midcingulate cortex, reflecting the decrease in subjective pain perception. Discussion: Our developed protocol allows for the investigation of short-term habituation with fMRI using self-report, block and trial-by-trial analyses and increases our understanding of individual differences in habitua-tion to pain. Process evaluation: Habituation can be measured in multiple ways, which complicates standardisation of paradigms and analyses. A protocol which allows for the integration and comparison of these methods helps to define and understand habituation. References: 1. Rankin CH, Abrams T, Barry RJ, et al. Habituation revisited: an updated and revised description of the behavioral characteristics of habituation. Neurobiology of learning and memory 2009; 92(2): 135-138. Introduction: The relevance ...