Simple SummaryIn 2017, 9.4 million animals were used for research and testing in the European Union. Animal testing always entails the potential for harm caused to the animals. In order to minimize animal suffering, it is of ethical and scientific interest to have a research-based severity assessment of animal experiments. In the past, many methods have been developed to investigate animal suffering. Initially, the focus was on physiological parameters, such as body weight or glucocorticoids as an indicator of stress. In addition, the animals’ behavior has come more into focus and has been included as an indicator of severity. However, in order to obtain a comprehensive understanding of animal suffering, an animal’s individual perspective should also be taken into account. Preference tests might be used, for example, to “ask” animals what they prefer, and providing such goods in turn allows, among other things, to improve housing conditions. In this review, different methods are introduced, which can be used to investigate and evaluate animal suffering and well-being with a special focus on animal-centric strategies.It has become mandatory for the application for allowance of animal experimentation to rate the severity of the experimental procedures. In order to minimize suffering related to animal experimentation it is therefore crucial to develop appropriate methods for the assessment of animal suffering. Physiological parameters such as hormones or body weight are used to assess stress in laboratory animals. However, such physiological parameters alone are often difficult to interpret and leave a wide scope for interpretation. More recently, behavior, feelings and emotions have come increasingly into the focus of welfare research. Tests like preference tests or cognitive bias tests give insight on how animals evaluate certain situations or objects, how they feel and what their emotional state is. These methods should be combined in order to obtain a comprehensive understanding of the well-being of laboratory animals.