The aim of the present study was to characterize participants’ abilities to answer binary yes/no questions by mentally manipulating imagery to produce imagined changes in luminance, which would in turn cause reflexive perturbations in pupil diameter. First, a paired association was established with participants, linking “yes” responses with imagining a “sunny sky” and “no” responses with imagining a “dark room”. Participants (N=20) then answered 16 yes/no questions using this response method, in place of providing verbal or gestural (e.g., head nod) answers. Pupil diameters were recorded for a period of 8000 ms following each stimulus question while participants maintained the mental image that corresponded with their answer. We hypothesized that on average, “no” responses would yield a pupil dilation and increased diameter relative to baseline, while “yes” responses would instead result in constrictions and smaller pupil diameters compared to baseline. A 2-factor repeated measures analysis of variance (ANOVA), where time was one factor and response type (i.e., yes or no) was the other, revealed a statistically significant interaction of time and response type, a significant main effect of time, and a trend toward significance for response type in aggregated group data. Item level discrimination consisted of comparing the mean pupil diameter in response to a single item for a single participant (e.g., “yes” response on one trial) to the mean pupil diameter of all contrasting responses for that same participant (e.g., all “no” response trials). This method achieved a 64.5% discrimination accuracy. This investigation affirmed the plausibility of leveraging pupillometry and luminance-based mental imagery in favor of an alternative communication system for individuals who are locked-in, as well as its potential as a screening tool. However, further investigation is warranted prior to its implementation. / Communication Sciences