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High-resolution whole-brain staining for electron microscopic circuit reconstruction.

Shawn Mikula1, Winfried Denk1

  • 1Department of Biomedical Optics, Max Planck Institute for Medical Research, Heidelberg, Germany.

Nature Methods
|April 14, 2015
PubMed
Summary

A new method called BROPA (brain-wide reduced-osmium staining with pyrogallol-mediated amplification) enables high-resolution, brain-wide ultrastructural preservation. This technique is suitable for reconstructing neuronal circuits across an entire mouse brain.

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Area of Science:

  • Neuroscience
  • Electron Microscopy
  • Neuroanatomy

Background:

  • Reconstructing neuronal circuits requires high resolution, typically achieved with electron microscopy.
  • Understanding brain-wide computations necessitates mapping neural circuits throughout the entire brain.

Purpose of the Study:

  • To develop a novel preparation method for preserving and staining ultrastructural details throughout the brain.
  • To enable the reconstruction of neural circuits at a single-synapse resolution across an entire brain.

Main Methods:

  • Developed BROPA (brain-wide reduced-osmium staining with pyrogallol-mediated amplification) for tissue preparation.
  • Utilized serial block-face electron microscopy (SBEM) for imaging.
  • Assessed human annotator ability to trace neural processes and identify synapses.

Main Results:

  • BROPA provides preservation and staining of ultrastructural details necessary for tracing neuronal processes.
  • The method allows for the identification of synaptic contacts.
  • Human annotators could reliably follow neural 'wires' over long distances using SBEM data from BROPA-prepared tissue.

Conclusions:

  • The BROPA method yields a preparation suitable for reconstructing neural circuits across an entire mouse brain.
  • This advancement facilitates large-scale connectomics research.