Related Experiment Video
Updated: Jan 18, 2026
08:57
Focussed Ion Beam Milling and Scanning Electron Microscopy of Brain Tissue
Published on: July 6, 2011
28.7K
Protocol for fixation and staining of postmortem human brain tissue optimized for focused ion beam-scanning electron
Matthew J Maier1, David A Lewis1, Zachary Freyberg2
1University of Pittsburgh, School of Medicine, Department of Psychiatry, Pittsburgh, PA 15213, USA.
STAR Protocols
|January 16, 2026
Summary
This study details a protocol for preparing postmortem human brain tissue for high-resolution volume electron microscopy (vEM) using focused ion beam-scanning electron microscopy (FIB-SEM). The method ensures excellent ultrastructural preservation for detailed 3D reconstructions.
Area of Science:
- Neuroscience
- Microscopy
- Biotechnology
Background:
- High-resolution imaging of postmortem human brain tissue is crucial for understanding neurological disorders.
- Existing protocols often struggle with preserving delicate ultrastructure for advanced microscopy techniques.
Purpose of the Study:
- To present a robust protocol for generating postmortem human brain tissue samples optimized for volume electron microscopy (vEM).
- To enable detailed three-dimensional (3D) ultrastructural reconstructions of human brain tissue.
Main Methods:
- Immersion fixation of brain tissue.
- Sectioning and long-term cryopreservation techniques.
- Enhanced-contrast staining for improved vEM imaging.
Main Results:
- The protocol yields postmortem human brain samples with high-quality ultrastructural preservation.
- Samples are suitable for high-resolution focused ion beam-scanning electron microscopy (FIB-SEM).
- Enables dense and targeted 3D reconstructions of cellular and subcellular features.
Conclusions:
- This protocol provides a reliable method for preparing human postmortem brain tissue for vEM.
- Facilitates detailed ultrastructural analysis and 3D reconstruction of the human brain.
- Advances research in neurobiology and neuropathology through improved imaging capabilities.
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