Objectives: Stroke predominantly affects the elderly. Universities of the Third Age (U3A) are presented with an opportunity to target them. The goal of our study was to improve older adults' preparedness to call 911 as a response to symptoms of stroke. Materials and Methods: Participants were recruited from U3A in Brno, Czech Republic in year 2018. The program included an educational movie about stroke and testing with pretest posttest design. Stroke awareness was measured by Stroke Action Test and video-clips portraying stroke and stroke mimicking symptoms. Respondents had to answer close-ended questions. Composite scores were com-pared using paired t-test. Results: Data were obtained from 206 attendees of the pro-gram, that is 2% of all students, from 4 of 5 U3A in Brno. The mean test score improved from 80% to 87% (paired p < 0.001). Participants with a lower baseline knowledge improved by 12% (95% CI 9% to 16%) and with a higher baseline knowledge by 0% (95% CI 3% to 4%). The score for calling 911 for stroke mimicking symptoms improved from 29% to 20% (paired p < 0.001). Conclusions: Video-based educational program improved senior preparedness to call 911 as a response to stroke. The improvement was mild, which is at least partly due to a high baseline level of preparedness of seniors active in U3A. A lower baseline knowledge was however associated with a bigger improvement, which might be important for use in a less active/educated population. Educational intervention also decreased inten-tion to call 911 for stroke mimicking symptoms, which could have important impli-cations for decreasing unnecessary activation of pre-hospital services.