This study assessed students’ perceptions of a ubiquitous 360° learning environment (ULE) in histotechnology comparing ULE perceptions to conventional web-based learning environment (WLE). Based on pervasive, seamless ubiquitous technologies and computing, ULE combines an authentic learning environment, i.e. histotechnology laboratory, with digital learning resources, embedded functional objects, mobile devices and sensing technologies. A highly usable user interface was created using a 360° spherical panorama imaging technique. The ULE was developed and conducted by a research group. Students’ perceptions on 360° ULE use were assessed in a quasi-experimental pilot study. From a histotechnology course, 57 out of 100 students participated voluntarily. The 360° ULE experimental group (n = 29) and WLE control group (n = 28) studied for 8 wks in 2014–2105. Assessment focused on the main criteria for ULE, i.e. flexibility, personalization, context-awareness, interactivity subscales. The WLE control group was assessed as more valuable from flexibility, context-awareness, and interactivity perspectives. Only the personalization aspect was perceived as higher in ULE than WLE. WLE group numerical values were systematically higher than the ULE group values, though statistically significant differences were revealed between groups. Both ULE and WLE environments supported learning. All measured subscales were assessed as high. Personalization and context-awareness aspects were evaluated as highest in both ULE and WLE groups, while interactivity was perceived as lowest. This study has implications for those developing technology-enhanced learning environments. Our developed 360° ULE could be easily implemented for health, medical, or nursing education as either practical training or continuing education.