Repeatable target localization for long-term in vivo imaging of mice with 2-photon microscopy

► A new head fixation device for repetitive in vivo imaging was designed. ► The new device allows fast, precise and automatic re-location to previously chosen brain volumes. ► Reduction of translational and rotational errors simplifies image post-processing. ► As laser exposure is limited to data acquisition, the risk of laser induced tissue damage and photobleaching is greatly reduced.