The assessment of pain, distress and suffering, as well as evaluation of the efficacy of stress-reduction strategies, is crucial in animal experimentation but can be challenging in laboratory mice. Nest building and burrowing performance, observed in the home cage, have proved to be valuable and easy-to-use tools to assess brain damage or malfunction as well as neurodegenerative diseases. Both behaviors are used as parameters in models of psychiatric disorders or to monitor sickness behavior following infection. Their use has been proposed in more realistic and clinically relevant preclinical models of disease, and reduction of these behaviors seems to be especially useful as an early sign of dysfunction and to monitor disease progression. Finally, both behaviors are reduced by pain and stress. Therefore, in combination with specific disease markers, changes in nest building and burrowing performance may help provide a global picture of a mouse`s state, and thus aid monitoring to ensure well-being in animal experimentation.