Hands-on dissections of animals are traditionally regarded as an essential part of biology education. Nowadays, regardless of the reported educational benefits, there is a gradual change in the concept and acceptance of hands-on dissection, leading in many cases to its abandonment in schools and its replacement with alternatives such as 3D models, figurines, plastination and computer-based alternatives. However, the position of hands-on dissection has recently been challenged, mostly by computer-supported alternatives. The aim of the study was to explore whether teacher self-efficacy can be recognized as a predictor of the application of the different kinds of animals in hands-on and virtual dissections in Biology lessons. Based on responses from 405 Czech Biology teachers there are differences in the acceptability and implementation of organisms for hands-on and virtual dissection. It was established, that self-efficacy is not a predictor of either the acceptability of organisms or actual behaviour in both variants of dissection.