Antonino Paolo Di Giovanna

University of Florence

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Francesco S. Pavone

University of Florence

Leonardo Sacconi

Istituto di Fisiologia Clinica

Ludovico Silvestri

University of Florence

Marie Caroline Müllenbroich

University of Glasgow

Irene Costantini

University of Florence

Anna Letizia Allegra Mascaro

National Research Council

Paolo Frasconi

University of Florence

Alessandro Panarese

University of Chieti-Pescara

Silvestro Micera

Scuola Superiore Sant'Anna

Claudia Alia

Neuroscience Institute

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University of Florence

National Research Council

National Academies of Sciences, Engineering, and Medicine

European Theoretical Spectroscopy Facility

National Institute of Optics

Instituto de Óptica "Daza de Valdés"

Florence (Netherlands)

Scuola Superiore Sant'Anna

Centre Hospitalier de Lens

Neuroscience Institute

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Whole-Brain Vasculature Reconstruction at the Single Capillary Level

Scientific Reports (2018)

Antonino Paolo Di Giovanna Alessandro Tibo Ludovico Silvestri Marie Caroline Müllenbroich Irene Costantini

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.

Connectomics

Vascular network

10.1038/s41598-018-30533-3

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Citations (111)

gel-BSA-FITC TPFM manual segmentation gel-BSA-FITC_manual_segmentation0242.tif

Harvard Dataverse (2018)

Antonino Paolo Di Giovanna

10.7910/dvn/hnls4b/qe73su

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gel-BSA-FITC labelled blood vessels Acquisition with TPFM gel-BSA-FITC_TPFM0091.tif