Naturalistic stimulation (i.e., movies and auditory narratives of some minutes’ length) has been a powerful approach to bringing more real-life experiences into laboratory experiments. Data-driven, intersubject correlation (ISC) analysis permits examining to what extent activity in a specific brain region correlates across participants during exposure to a naturalistic stimulus, as well as testing whether neural activity correlates with behavioral measures. Notably, most of the previous research with naturalistic stimuli was conducted using functional fMRI (fMRI). Here, we tested whether a naturalistic approach and the ISC are feasible using functional near-infrared spectroscopy (fNIRS) - the imaging method particularly suited for populations of patients and children. Fifty-three healthy adult participants watched twice a 3-min segment of a Charlie Chaplin movie while we recorded the brain activity on the surface of their prefrontal cortex using fNIRS. In addition, an independent group of 18 participants used a continuous scoring procedure to rate the extent to which they felt that different parts of the movie fragment were funny. Our two findings were as follows. First, we found higher-than-zero ISC in fNIRS signals in the prefrontal cortex lobes, a result that was particularly high in the oxygenated channels during the first repetition of the movie. Second, we found a significant negative correlation between oxygenated brain signals and ratings of the movie’s humorousness. In a series of control analyses we demonstrated that this latter correlation could not be explained by various non-humor-related movie sensory properties (e.g., auditory volume and image brightness). The key overall outcome of the present study is that fNIRS in combination with the naturalistic paradigms and the ISC might be a sensitive and powerful research method to explore cognitive processing. Our results also suggest a potential role of the prefrontal cortex in humor appreciation.