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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 17, 2026

Preparation of Mouse Brain Tissue for Immunoelectron Microscopy

Preparation of Mouse Brain Tissue for Immunoelectron Microscopy

Published on: July 20, 2010

Rehydration of Freeze Substituted Brain Tissue for Pre-embedding Immunoelectron Microscopy.

Janeth Pérez-Garza¹, Emily Parrish-Mulliken¹, Zachary Deane¹

¹Department of Physiology and Neurobiology, University of Connecticut, 75 North Eagleville Rd. Unit 3156, Storrs, CT 06269-3156, USA.

Microscopy and Microanalysis : the Official Journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada

|August 16, 2023

Summary

We developed a cryofixation method for electron microscopy (EM) volume reconstruction. This technique enables sensitive molecular labeling while preserving ultrastructure for detailed brain circuit analysis.

Keywords:

EM volume reconstruction antibody labeling electron microscopy freeze substitution high-pressure freezing neuroanatomy

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Last Updated: Jun 17, 2026

Preparation of Mouse Brain Tissue for Immunoelectron Microscopy

Preparation of Mouse Brain Tissue for Immunoelectron Microscopy

Published on: July 20, 2010

Preparation of Non-human Primate Brain Tissue for Pre-embedding Immunohistochemistry and Electron Microscopy

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Published on: April 3, 2017

Free-floating Immunostaining of Mouse Brains

Free-floating Immunostaining of Mouse Brains

Published on: October 7, 2021

Area of Science:

Neuroscience
Cell Biology
Microscopy

Background:

Electron microscopy (EM) volume reconstruction is vital for studying brain circuits.
Integrating molecular data with EM is challenging due to morphological compromises from current labeling methods.
High-quality ultrastructural preservation and contrasted cell membranes are crucial for EM reconstruction.

Purpose of the Study:

To develop a novel method for combining sensitive molecular labeling with high-fidelity ultrastructural preservation for EM volume reconstruction.
To overcome the limitations of existing antibody labeling techniques that degrade cellular morphology.

Main Methods:

Utilized cryofixation for superior morphological preservation of biological samples.
Developed a cryofixation-based protocol for sensitive immunolabeling of endogenous molecules.
Ensured compatibility with high-contrast staining techniques required for serial EM reconstruction.

Main Results:

Achieved excellent ultrastructural preservation using the cryofixation method.
Demonstrated sensitive immunolabeling of endogenous molecules without compromising morphology.
Validated compatibility with high-contrast staining for serial EM analysis.

Conclusions:

The developed cryofixation method enhances EM volume reconstruction by enabling integrated molecular and ultrastructural analysis.
This technique overcomes previous limitations, allowing for more comprehensive studies of brain circuitry.
The method preserves valuable samples and facilitates detailed neuronal process tracing.