High-speed imaging with light-sheet fluorescence microscopy poses several challenges throughout the whole pipeline, from data acquisition in the lab to image stitching and post-processing. Here we present our custom hardware and software solutions that allow us to map large biological samples at the cellular level, e.g. large portions of human brain cortex. Our custom optical setup—a dual-view, inverted, light-sheet microscope—is capable of simultaneous two-color acquisition at a data rate of 1 GB/s. Our open source tools include the instrument’s data acquisition and control software and also cover volumetric image stitching and post-processing.
3D reconstruction of the human brain at high resolution is one of the most important challenges of neuroscience. We present a new clearing method named SHORT that in combination with an advanced double-view light-sheet fluorescence microscope and an automated machine-learning based images analysis allow to perform volumetric study of the human brain. We applied our methodology to a Broca’s area block of 4 x 4 x 2 cm3, demonstrating the possibility of obtaining a fast 3D reconstruction of the human brain at high-resolution, paving the way to the possibility of finally mapping a comprehensive atlas of the human brain.
This conference presentation, “3D molecular phenotyping of the human brain Broca’s area using light-sheet fluorescence microscopy” was prepared for the Biomedical Spectroscopy, Microscopy, and Imaging II conference at SPIE Photonics Europe 2022.
We still lack a detailed map of the anatomical disposition of neurons in the human brain. A complete map would be an important step for deeply understanding the brain function, providing anatomical information useful to decipher the neuronal pattern in healthy and diseased conditions. Here, we present several important advances towards this goal, obtained by combining a new clearing method, advanced Light Sheet Microscopy and automated machine-learning based image analysis. We perform volumetric imaging of large sequentially stained human brain slices, labelled for two different neuronal markers NeuN and GAD67, discriminating the inhibitory population and reconstructing the brain connectivity.
We still lack a detailed map of the anatomical disposition of neurons in the human brain. A complete map would be an important step for deeply understanding the brain function, providing anatomical information useful to decipher the neuronal pattern in healthy and diseased conditions. Here, we present several important advances towards this goal, obtained by combining a new clearing method, advanced Light Sheet Microscopy and automated machine-learning based image analysis. We perform volumetric imaging of large sequentially stained human brain slices, labelled for two different neuronal markers NeuN and GAD67, discriminating the inhibitory population and reconstructing the brain connectivity.
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