A platform for brain-wide imaging and reconstruction of individual neurons

ELife
Michael N. EconomoJayaram Chandrashekar

Abstract

The structure of axonal arbors controls how signals from individual neurons are routed within the mammalian brain. However, the arbors of very few long-range projection neurons have been reconstructed in their entirety, as axons with diameters as small as 100 nm arborize in target regions dispersed over many millimeters of tissue. We introduce a platform for high-resolution, three-dimensional fluorescence imaging of complete tissue volumes that enables the visualization and reconstruction of long-range axonal arbors. This platform relies on a high-speed two-photon microscope integrated with a tissue vibratome and a suite of computational tools for large-scale image data. We demonstrate the power of this approach by reconstructing the axonal arbors of multiple neurons in the motor cortex across a single mouse brain.

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Methods Mentioned

BETA
volumetric microscopy
fluorescence imaging
fluorescence microscopy
light scattering
Fluorescence
light microscopy
electron microscopy
transmission electron microscopy
electron

Software Mentioned

CUBIC
Fiji
RegularizeData3d
Matlab
libavcodec
Janelia Workstation

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