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High-resolution whole-brain staining for electron microscopic circuit reconstruction.
1Department of Biomedical Optics, Max Planck Institute for Medical Research, Heidelberg, Germany.
Nature Methods
|April 14, 2015
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.
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.
