Functional and multiscale 3D structural investigation of brain tissue through correlative in vivo physiology, synchrotron microtomography and volume electron microscopy

• Published in: Nature communications Vol. 13; no. 1; pp. 2923 - 16
• Main Authors: Bosch, Carles, Ackels, Tobias, Pacureanu, Alexandra, Zhang, Yuxin, Peddie, Christopher J., Berning, Manuel, Rzepka, Norman, Zdora, Marie-Christine, Whiteley, Isabell, Storm, Malte, Bonnin, Anne, Rau, Christoph, Margrie, Troy, Collinson, Lucy, Schaefer, Andreas T.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 25.05.2022; Nature Publishing Group; Nature Portfolio
• Subjects: 14/69; 147/135; 631/378/2597/2600; 631/378/2624/1703; 631/378/3920; 64/60; Animal tissues; Animals; Brain; Brain - diagnostic imaging; Brain - ultrastructure; Calcium imaging; Computed tomography; Context; Dehydration; Diffusion Magnetic Resonance Imaging; Electron microscopy; Humanities and Social Sciences; Imaging, Three-Dimensional; In vivo methods and tests; Mice; Microscopy; Microscopy, Electron; Microscopy, Electron, Scanning; Microtomography; multidisciplinary; Neuroimaging; Olfactory bulb; Olfactory pathways; Phase contrast; Physiology; Pyramidal cells; Science; Science (multidisciplinary); Spine; Structure-function relationships; Synchrotron radiation; Synchrotrons; Tissues; X-Ray Microtomography - methods
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-022-30199-6

Understanding the function of biological tissues requires a coordinated study of physiology and structure, exploring volumes that contain complete functional units at a detail that resolves the relevant features. Here, we introduce an approach to address this challenge: Mouse brain tissue sections containing a region where function was recorded using in vivo 2-photon calcium imaging were stained, dehydrated, resin-embedded and imaged with synchrotron X-ray computed tomography with propagation-based phase contrast (SXRT). SXRT provided context at subcellular detail, and could be followed by targeted acquisition of multiple volumes using serial block-face electron microscopy (SBEM). In the olfactory bulb, combining SXRT and SBEM enabled disambiguation of in vivo-assigned regions of interest. In the hippocampus, we found that superficial pyramidal neurons in CA1a displayed a larger density of spine apparati than deeper ones. Altogether, this approach can enable a functional and structural investigation of subcellular features in the context of cells and tissues. The function of biological tissues is encoded in their physiology and structure. Here, Bosch et al. have integrated both insights to study specific neuronal circuits by combining in vivo light, synchrotron X-ray and volume electron microscopy.