Different online courses and training programs in Laboratory Animal Science (LAS) have emerged across Europe in recent years. E-learning appears to be a promising solution to achieve flexibility in training while meeting the quality criteria of demanding programs in short training periods. However, little is known about how students perceive e-learning in this context, and there is also a lack of specific and valid instruments to measure this perception. Within an exploratory study framework, the e-learning perception of 229 participants in 15 courses in Portugal using two different online training formats, flipped classroom and full online theoretical training, was assessed. For this purpose, the Questionnaire of E-learning Acceptance (QELA), a 32-item accordance Likert-type scale comprising five subscales was developed to explore the following: how participant perceive e-learning, satisfaction with organization and contents, perception of e-learning relevance for the time management, and its influence for practical training. In general, e-learning was well accepted and perceived to work well and be useful by the majority of courses participants, independently of the course level and e-learning format approach. These results indeed suggest that integration of e-learning is useful in LAS training. We also propose the QELA as a starting point for development and implementation of specific instruments to assess e-learning acceptance in LAS across a wider range of geographical and training contexts.