Dense neuronal reconstruction through X-ray holographic nano-tomography

Nat Neurosci. 2020 Dec;23(12):1637-1643. doi: 10.1038/s41593-020-0704-9. Epub 2020 Sep 14.

Abstract

Imaging neuronal networks provides a foundation for understanding the nervous system, but resolving dense nanometer-scale structures over large volumes remains challenging for light microscopy (LM) and electron microscopy (EM). Here we show that X-ray holographic nano-tomography (XNH) can image millimeter-scale volumes with sub-100-nm resolution, enabling reconstruction of dense wiring in Drosophila melanogaster and mouse nervous tissue. We performed correlative XNH and EM to reconstruct hundreds of cortical pyramidal cells and show that more superficial cells receive stronger synaptic inhibition on their apical dendrites. By combining multiple XNH scans, we imaged an adult Drosophila leg with sufficient resolution to comprehensively catalog mechanosensory neurons and trace individual motor axons from muscles to the central nervous system. To accelerate neuronal reconstructions, we trained a convolutional neural network to automatically segment neurons from XNH volumes. Thus, XNH bridges a key gap between LM and EM, providing a new avenue for neural circuit discovery.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Axons / physiology
  • Axons / ultrastructure
  • Cerebral Cortex / cytology
  • Cerebral Cortex / physiology
  • Cerebral Cortex / ultrastructure
  • Dendrites / physiology
  • Dendrites / ultrastructure
  • Drosophila melanogaster
  • Female
  • Holography
  • Image Processing, Computer-Assisted / methods*
  • Imaging, Three-Dimensional
  • Machine Learning
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Motor Neurons / physiology
  • Motor Neurons / ultrastructure
  • Muscle, Skeletal / innervation
  • Muscle, Skeletal / ultrastructure
  • Nanotechnology
  • Neural Networks, Computer
  • Neurons / ultrastructure*
  • Pyramidal Cells / ultrastructure
  • Sensory Receptor Cells / physiology
  • Sensory Receptor Cells / ultrastructure
  • Tomography