Quantifying Mesoscale Neuroanatomy Using X-Ray Microtomography

• Published in: eNeuro Vol. 4; no. 5; p. ENEURO.0195-17.2017
• Main Authors: Dyer, Eva L, Gray Roncal, William, Prasad, Judy A, Fernandes, Hugo L, Gürsoy, Doga, De Andrade, Vincent, Fezzaa, Kamel, Xiao, Xianghui, Vogelstein, Joshua T, Jacobsen, Chris, Körding, Konrad P, Kasthuri, Narayanan
• Format: Journal Article
• Language: English
• Published: United States Society for Neuroscience 01.09.2017
• Subjects: 60 APPLIED LIFE SCIENCES; Animals; automated segmentation; Blood Vessels - anatomy & histology; Blood Vessels - diagnostic imaging; Brain - anatomy & histology; Brain - diagnostic imaging; cell counting; electron microscopy; Female; Imaging, Three-Dimensional; MATHEMATICS AND COMPUTING; Methods/New Tools; Mice, Inbred BALB C; Myelin Sheath; neocortex; neuroanatomy; Neurons - cytology; Pattern Recognition, Automated; Reproducibility of Results; Software; X-ray microtomography; X-Ray Microtomography - instrumentation; X-Ray Microtomography - methods; electron microscopy; neocortex; cell counting; neuroanatomy; Automated segmentation; X-ray microtomography
• ISSN: 2373-2822; 2373-2822
• DOI: 10.1523/ENEURO.0195-17.2017

Methods for resolving the three-dimensional (3D) microstructure of the brain typically start by thinly slicing and staining the brain, followed by imaging numerous individual sections with visible light photons or electrons. In contrast, X-rays can be used to image thick samples, providing a rapid approach for producing large 3D brain maps without sectioning. Here we demonstrate the use of synchrotron X-ray microtomography (µCT) for producing mesoscale (∼1 µm resolution) brain maps from millimeter-scale volumes of mouse brain. We introduce a pipeline for µCT-based brain mapping that develops and integrates methods for sample preparation, imaging, and automated segmentation of cells, blood vessels, and myelinated axons, in addition to statistical analyses of these brain structures. Our results demonstrate that X-ray tomography achieves rapid quantification of large brain volumes, complementing other brain mapping and connectomics efforts.