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The promise of using immersive technologies in learning has increasingly been attracting researchers' and practitioners' attention. However, relevant empirical works are usually conducted in fully controlled Virtual Reality (VR) laboratories, as opposed to conventional settings. This quasi‐experimental study compares the effectiveness of video learning resources to that of stereoscopic 360° VR, as supplements to the traditional instructional approach. The potential of such methods was examined in high school settings, in the context of the 'Life and Evolution' module, with participants (n = 70) divided equally into control and experimental groups. As a point of reference (control condition), we considered the adoption of Video Learning Resources, as students are more acquainted with this instructional method. In the intervention approach (experimental condition), students adopted the use of low‐end mobile‐VR (VeeR Mini VR Goggles). The key findings indicate differences in the learning motivation, confidence and satisfaction, but no statistically significant difference was identified regarding the factual or conceptual knowledge gains. The study offers insights on the potential of the investigated technologies in the subject of secondary school Biology and further provides implications for theory and practice. Practitioner notesWhat is already known about this topicResearchers' interest over the potential of Virtual Reality on different STEM disciplines is increasing consistently.An increasing number of efforts can be identified discussing the integration of multimedia learning resources in the secondary school context.Empirical studies on the subject of Biology are focusing on students' academic performance and achievement but not on learning motivation and satisfaction.What this paper addsThis quasi‐experimental study comparatively examines academic performance, with the focus being on learning motivation and satisfaction, across different modalities (stereoscopic 360° Virtual Reality applications‐VR, Video Learning Recourses‐VLR).The findings demonstrate that both instructional methods are sufficient in enhancing students' knowledge acquisition and academic performance.The adoption of stereoscopic 360° VR influences students' learning motivation and impacts long‐term memory retention.Implications for practice and policyEducators are advised to consider the systematic adoption of "immersive" multimedia tools to enhance the subject of Biology as they can greatly encourage scientific inquiry.Instructional designers are advised to adopt open educational resources aligned to the curriculum of the local context.Educational researchers are advised to integrate stereoscopic 360°‐VR solutions in the conventional classroom settings. [ABSTRACT FROM AUTHOR]