%0 Journal Article %A Norio Takata %A Nobuhiko Sato %A Yuji Komaki %A Hideyuki Okano %A Kenji F. Tanaka %T Flexible annotation atlas of the mouse brain: combining and dividing brain structures of the Allen Brain Atlas while maintaining anatomical hierarchy %D 2020 %R 10.1101/2020.02.17.953547 %J bioRxiv %P 2020.02.17.953547 %X A brain atlas is necessary for analyzing structure and function in neuroimaging research. Although various annotation volumes (AVs) for the mouse brain have been proposed, it is common in magnetic resonance imaging (MRI) of the mouse brain that regions-of-interest (ROIs) for brain structures (nodes) are created arbitrarily according to each researcher’s necessity, leading to inconsistent ROIs among studies. One reason for such a situation is the fact that earlier AVs were fixed, i.e. combination and division of nodes were not implemented. This report presents a pipeline for constructing a flexible annotation atlas (FAA) of the mouse brain by leveraging public resources of the Allen Institute for Brain Science on brain structure, gene expression, and axonal projection. A mere two-step procedure with user-specified, text-based information and Python codes constructs FAA with nodes which can be combined or divided objectively while maintaining anatomical hierarchy of brain structures. Four FAAs with total node count of 4, 101, 866, and 1,381 were demonstrated. Unique characteristics of FAA realized analysis of resting-state functional connectivity (FC) across the anatomical hierarchy and among cortical layers, which were thin but large brain structures. FAA can improve the consistency of whole brain ROI definition among laboratories by fulfilling various requests from researchers with its flexibility and reproducibility.Highlights– A flexible annotation atlas (FAA) for the mouse brain is proposed.– FAA is expected to improve whole brain ROI-definition consistency among laboratories.– The ROI can be combined or divided objectively while maintaining anatomical hierarchy.– FAA realizes functional connectivity analysis across the anatomical hierarchy.– Codes for FAA reconstruction is available at https://github.com/ntakata/flexible-annotation-atlas– Datasets for resting-state fMRI in awake mice are available at https://openneuro.org/datasets/ds002551 %U https://www.biorxiv.org/content/biorxiv/early/2020/02/28/2020.02.17.953547.full.pdf