Objective: This research investigates the usability of image-based CAPTCHAs using performance, eye-gaze and subjective measures. Background: Several forms of image-based CAPTCHAs have recently been developed. Few studies have been conducted to examine their usability. Method: Twenty participants received training using four different CAPTCHAs: Asirra, ESP-PIX, SQ-PIX and IMAGINATION. The participants were then asked to complete the CAPTCHA challenge using five CAPTCHAs of each type. The participants were subsequently asked to complete the computer system usability questionnaire (CSUQ) and the NASA Task Load Index test. Upon completing all the tasks, the participants completed a final post-test subjective questionnaire ranking the CAPTCHAs based on preference. Results: Statistically significant differences among the CAPTCHAs were found for all dependent variables other than task completion time. Conclusion: Participants preferred Asirra and ESP-PIX to the other two CAPTCHAs.