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26 April 2016 Mapping whole-brain activity with cellular resolution by light-sheet microscopy and high-throughput image analysis (Conference Presentation)
Ludovico Silvestri, Nikita Rudinskiy, Marco Paciscopi, Marie Caroline Müllenbroich, Irene Costantini, Leonardo Sacconi, Paolo Frasconi, Bradley T. Hyman, Francesco S. Pavone
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Proceedings Volume 9690, Clinical and Translational Neurophotonics; Neural Imaging and Sensing; and Optogenetics and Optical Manipulation; 969012 (2016) https://doi.org/10.1117/12.2208603
Event: SPIE BiOS, 2016, San Francisco, California, United States
Abstract
Mapping neuronal activity patterns across the whole brain with cellular resolution is a challenging task for state-of-the-art imaging methods. Indeed, despite a number of technological efforts, quantitative cellular-resolution activation maps of the whole brain have not yet been obtained. Many techniques are limited by coarse resolution or by a narrow field of view. High-throughput imaging methods, such as light sheet microscopy, can be used to image large specimens with high resolution and in reasonable times. However, the bottleneck is then moved from image acquisition to image analysis, since many TeraBytes of data have to be processed to extract meaningful information. Here, we present a full experimental pipeline to quantify neuronal activity in the entire mouse brain with cellular resolution, based on a combination of genetics, optics and computer science. We used a transgenic mouse strain (Arc-dVenus mouse) in which neurons which have been active in the last hours before brain fixation are fluorescently labelled. Samples were cleared with CLARITY and imaged with a custom-made confocal light sheet microscope. To perform an automatic localization of fluorescent cells on the large images produced, we used a novel computational approach called semantic deconvolution. The combined approach presented here allows quantifying the amount of Arc-expressing neurons throughout the whole mouse brain. When applied to cohorts of mice subject to different stimuli and/or environmental conditions, this method helps finding correlations in activity between different neuronal populations, opening the possibility to infer a sort of brain-wide 'functional connectivity' with cellular resolution.
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Ludovico Silvestri, Nikita Rudinskiy, Marco Paciscopi, Marie Caroline Müllenbroich, Irene Costantini, Leonardo Sacconi, Paolo Frasconi, Bradley T. Hyman, and Francesco S. Pavone "Mapping whole-brain activity with cellular resolution by light-sheet microscopy and high-throughput image analysis (Conference Presentation)", Proc. SPIE 9690, Clinical and Translational Neurophotonics; Neural Imaging and Sensing; and Optogenetics and Optical Manipulation, 969012 (26 April 2016); https://doi.org/10.1117/12.2208603
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KEYWORDS
Brain
Brain mapping
Image resolution
Image analysis
Microscopy
Image processing
Neurons
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