In contrast to human brain imaging, study-independent templates are rarely available in preclinical settings, limiting comparability and the use of common brain coordinates. Furthermore, most available studies are restricted to male mice, hindering the use of available data for comparative evaluation in projects including female animals. To balance this shortcoming and provide a resource for future studies, to enhance comparability and reduce animal numbers, we present a novel, in-vivo brain template derived from a large, exclusively female cohort of adult C57Bl6/N mice.80 treatment-naïve, female C57Bl6/N mice (Charles River, Germany), aged 12-15 weeks, underwent in-vivo magnetic resonance imaging (MRI) under general anesthesia (isoflurane, medetomidine) as baseline measurements for a longitudinal study. All experimental procedures were approved by national authorities and conducted adhering to the ARRIVE guideline. Measurements were performed on a 9.4T Bruker BioSpec® 94/30 USR μMRI scanner using a helium-cooled CryoProbe™.The structural, T2-weighted MR scans (TurboRARE) were spatially aligned, bias-corrected and semi-automatically masked in MATLAB using SPM12 and Turone Mouse Brain Atlas and Template (TMBTA) tissue priors. After visual inspection and mask refinement (3D Slicer) or exclusion of scans, a 60µm-isotropic brain template was created with the software package Advanced Normalization Tools from 77 axial (75x250x75µm) and 76 coronal (78x38x500µm) scans.The resulting FeBls template brain (see Figure 1) was stripped of any remaining non-brain tissue and will be made available to the public via the NeuroImaging Tools and Resources Collaboratory (nitrc.org).