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Preparation of Samples for Electron Microscopy

Preparation of Samples for Electron Microscopy

To be visualized by an electron microscope, either transmission or scanning, biological samples need to be fixed (stabilized) so the electron beam does not destroy them and dried thoroughly (desiccated/dehydrated) so the vacuum does not affect them. Fixation needs to be done as quickly as possible because the sample properties will start changing as soon as it is removed from its natural environment. For example, in a tissue sample, the oxygen levels begin decreasing, causing an altered...

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Related Experiment Video

Updated: Jun 28, 2025

Preparation of Mouse Brain Tissue for Immunoelectron Microscopy

Preparation of Mouse Brain Tissue for Immunoelectron Microscopy

Published on: July 20, 2010

Mouse Brain Tissue Preparation for Scanning Electron Microscopy.

Kosuke Mori¹, Chihiro Takada², Hideshi Okada²

¹Departments of Tumor Pathology, Gifu University Graduate School of Medicine, Gifu, Japan.

Methods in Molecular Biology (Clifton, N.J.)

|April 17, 2024

Summary

Scanning electron microscopy (SEM) offers detailed 3D surface imaging. A new fixation and freeze-cracking method enables high-resolution ultrastructure observation in water-rich samples like mouse brains.

Keywords:

Freeze-cracking Freeze-drying Perfusion fixation Scanning electron microscope

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Last Updated: Jun 28, 2025

Preparation of Mouse Brain Tissue for Immunoelectron Microscopy

Preparation of Mouse Brain Tissue for Immunoelectron Microscopy

Published on: July 20, 2010

Specimen Preparation, Imaging, and Analysis Protocols for Knife-edge Scanning Microscopy

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Published on: December 9, 2011

Micron-scale Resolution Optical Tomography of Entire Mouse Brains with Confocal Light Sheet Microscopy

Micron-scale Resolution Optical Tomography of Entire Mouse Brains with Confocal Light Sheet Microscopy

Published on: October 8, 2013

Area of Science:

Microscopy
Biological Imaging
Neuroscience

Background:

Scanning electron microscopy (SEM) is crucial for observing surface topography with high resolution and large depth of field.
Conventional SEM methods face challenges in preserving the ultrastructure of hydrated biological tissues.
Freeze-cracking offers a pathway to visualize internal structures, but requires optimized sample preparation.

Purpose of the Study:

To present an optimized fixation and freeze-cracking protocol for mouse brain samples.
To enable high-resolution imaging of ultrastructure in hydrated biological specimens using SEM.
To overcome limitations in observing delicate biological samples with SEM.

Main Methods:

Sample preparation involving chemical fixation of mouse brain tissue.
Cryo-fracturing of fixed tissue to expose internal structures.
Post-fracture processing and sputter-coating for SEM observation.
Utilizing Scanning Electron Microscopy (SEM) for high-resolution imaging.

Main Results:

Successful preservation of fine ultrastructural details in mouse brain tissue after fixation and freeze-cracking.
Demonstration of the three-dimensional morphology of cellular and extracellular components.
High-resolution imaging of cross-sections, revealing tissue architecture.

Conclusions:

The described fixation and freeze-cracking method is effective for SEM analysis of mouse brain ultrastructure.
This technique significantly improves the ability to study hydrated biological samples at the nanoscale.
The protocol facilitates detailed investigation of tissue morphology and cellular organization.