High-resolution synchrotron-based X-ray microtomography as a tool to unveil the three-dimensional neuronal architecture of the brain

Sci Rep. 2018 Aug 13;8(1):12074. doi: 10.1038/s41598-018-30501-x.

Abstract

The assessment of neuronal number, spatial organization and connectivity is fundamental for a complete understanding of brain function. However, the evaluation of the three-dimensional (3D) brain cytoarchitecture at cellular resolution persists as a great challenge in the field of neuroscience. In this context, X-ray microtomography has shown to be a valuable non-destructive tool for imaging a broad range of samples, from dense materials to soft biological specimens, arisen as a new method for deciphering the cytoarchitecture and connectivity of the brain. In this work we present a method for imaging whole neurons in the brain, combining synchrotron-based X-ray microtomography with the Golgi-Cox mercury-based impregnation protocol. In contrast to optical 3D techniques, the approach shown here does neither require tissue slicing or clearing, and allows the investigation of several cells within a 3D region of the brain.

MeSH terms

  • Animals
  • Brain / cytology*
  • Brain / diagnostic imaging
  • Imaging, Three-Dimensional / instrumentation
  • Imaging, Three-Dimensional / methods*
  • Mercuric Chloride / chemistry
  • Mice
  • Neurons*
  • Silver Staining / methods
  • Synchrotrons
  • Tissue Fixation / methods
  • X-Ray Microtomography / instrumentation
  • X-Ray Microtomography / methods*

Substances

  • Mercuric Chloride