Whole-Brain Vasculature Reconstruction at the Single Capillary Level

Year: 2018

Authors: Di Giovanna A.P., Tibo A., Silvestri L., Muellenbroich M.C., Costantini I., Allegra Mascaro A.L., Sacconi L., Frasconi P., Pavone F.S.

Autors Affiliation: European Laboratory for Non-Linear Spectroscopy, University of Florence, Via Nello Carrara 1, Sesto Fiorentino, 50019, Italy; Department of Information Engineering (DINFO), University of Florence, Via di S. Marta 3, Florence, 50139, Italy; National Institute of Optics, National Research Council, Largo Fermi 6, Florence, 50125, Italy; Neuroscience Institute, National Research Council, Via Giuseppe Moruzzi 1, Pisa, 56125, Italy; Department of Physics and Astronomy, University of Florence, Via Sansone 1, Sesto Fiorentino, 50019, Italy

Abstract: The distinct organization of the brain’s vascular network ensures that it is adequately supplied with oxygen and nutrients. However, despite this fundamental role, a detailed reconstruction of the brain-wide vasculature at the capillary level remains elusive, due to insufficient image quality using the best available techniques. Here, we demonstrate a novel approach that improves vascular demarcation by combining CLARITY with a vascular staining approach that can fill the entire blood vessel lumen and imaging with light-sheet fluorescence microscopy. This method significantly improves image contrast, particularly in depth, thereby allowing reliable application of automatic segmentation algorithms, which play an increasingly important role in high-throughput imaging of the terabyte-sized datasets now routinely produced. Furthermore, our novel method is compatible with endogenous fluorescence, thus allowing simultaneous investigations of vasculature and genetically targeted neurons. We believe our new method will be valuable for future brain-wide investigations of the capillary network.

Journal/Review: SCIENTIFIC REPORTS

Volume: 8      Pages from: 12573-1  to: 12573-11

More Information: We gratefully acknowledge Giacomo Mazzamuto from the European Laboratory for Non-Linear Spectroscopy for converting the original whole-brain dataset in a compressed format, suitable for uploading on the online repository. This project received funding from the European Union’s H2020 research and innovation programme under grant agreements No. 720270 (Human Brain Project) and 654148 (Laserlab-Europe), and from the EU programme H2020 EXCELLENT SCIENCE – European Research Council (ERC) under grant agreement ID n. 692943 (BrainBIT). The project have also been supported by the Italian Ministry for Education, University, and Research in the framework of the Flagship Project NanoMAX and of Eurobioimaging Italian Nodes (ESFRI research infrastructure), and by Ente Cassa di Risparmio di Firenze (private foundation).
KeyWords: Light sheet microscopy; Magnetic-resonance angiography; Mouse-brain; Blood-flow; Clearing agent; Visualization; Tissue; Ultramicroscopy; Colocalization; Tomography; Brain blood vessel; Fluorescence microscopy; Nerve cell
DOI: 10.1038/s41598-018-30533-3

Citations: 85
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