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Related Experiment Video

Updated: Jul 14, 2025

Wholemount Immunohistochemistry for Revealing Complex Brain Topography
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A Scalable Staining Strategy for Whole-Brain Connectomics.

Xiaotang Lu1, Yuelong Wu1, Richard L Schalek1

  • 1Department of Molecular and Cellular Biology and The Center for Brain Science, Harvard University, Cambridge, Massachusetts, 02138, USA.

Biorxiv : the Preprint Server for Biology
|October 9, 2023
PubMed
Summary

Developing a new protocol, ODeCO, enables complete synaptic mapping of mouse brains. This method preserves ultrastructure and tissue integrity for comprehensive connectome construction.

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Area of Science:

  • Neuroscience
  • Connectomics
  • Electron Microscopy

Background:

  • Mapping mammalian brain synaptic connectivity is crucial for understanding perception, behavior, and memory.
  • Serial section electron microscopy is vital for visualizing neural connections but faces challenges in whole brains.
  • Chemical treatments and volume changes can compromise ultrastructure and tissue integrity.

Approach:

  • Developed a 12-step protocol, ODeCO (Osmium Diffusion and Clearing Optimization).
  • Utilized time-lapse X-ray imaging and brain proxies for optimized osmium infiltration.
  • Ensured uniform osmium penetration throughout the entire mouse brain.

Key Points:

  • ODeCO protocol effectively infiltrates osmium tetroxide across whole mouse brains.
  • Preserves ultrastructural quality and macroscopic tissue integrity, preventing cracks and fragmentation.
  • Overcomes key challenges in preparing large-scale neural tissue for electron microscopy.

Conclusions:

  • The ODeCO protocol is a prerequisite for constructing the first comprehensive mouse brain connectome.
  • Enables high-resolution mapping of neural pathways at the synaptic level.
  • Advances the field of connectomics for mammalian brain research.