Combined three-dimensional, haptic-enabled, virtual reality (3D HE VR) systems allow students to actively engage and explore various science concepts by leveraging user-friendly and immersive interfaces. Successful implementation of these learning tools in science classrooms hinges upon teachers’ perceptions of the technology’s potential as a viable pedagogical tool. Prior studies using the Technology Acceptance (TA) Model (TAM) suggest pre-service teachers have greater TA compared to in-service teachers. This study sought to explore how 3D HE VR designed to diminish Ease of Use (EOU) issues, influenced TA (through reported preferences) between pre-service and in-service science teachers. Five pre-service and five in-service teachers reported Perceived Utility (PU) and EOU upon using a 3D HE VR system (zSpace®) to learn science concepts. Quantitative data were collected from pre- and post-test content assessments. Qualitative data were collected and transcribed from field notes and interviews. Both teacher groups evidenced learning gains and reported EOU using zSpace®. However, preference for the technology compared to traditional methods varied between teacher groups. Sampled pre-service teachers held a significant preference for hands-on activities for instruction whereas in-service teachers reported greater TA, citing its potential to increase student interest in science and opportunity for personalized learning. This research suggests that when perceived EOU is mitigated, PU may more readily mediate TA among in-service teachers as they can envision the use of 3D HE VR technology use in teaching practices. Further exploration is needed to leverage in-service teachers’ classroom experience to implement novel forms of technology into their science instruction.