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Updated: Jun 8, 2026

A Hydrophobic Tissue Clearing Method for Rat Brain Tissue

A Hydrophobic Tissue Clearing Method for Rat Brain Tissue

Published on: December 23, 2020

Rapidly excised and cryofixed rat tissue.

Dimitri Vanhecke¹, Werner Graber, Daniel Studer

¹Institute of Anatomy, University of Berne, CH-3000 Bern 9, Switzerland.

Methods in Cell Biology

|September 28, 2010

Summary

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High-pressure freezing of microbiopsies offers a superior alternative to chemical fixation for preserving biological sample ultrastructure. This method minimizes artifacts, ensuring more accurate electron microscopy of tissues like brain, liver, kidney, and muscle.

Area of Science:

Electron microscopy
Cell biology
Biotechnology

Background:

Preserving native biological structures is critical for electron microscopy.
Chemical fixation methods like perfusion and excision introduce artifacts due to slow diffusion and osmotic effects.
Existing methods often result in cell swelling or shrinkage, compromising ultrastructural integrity.

Purpose of the Study:

To introduce microbiopsy and high-pressure freezing/freeze-substitution (HPF/FS) as an improved sample preparation technique.
To overcome the limitations of chemical fixation in preserving cellular ultrastructure.
To provide a rapid and effective method for obtaining high-quality biological samples for microscopy.

Main Methods:

Utilizing microbiopsies for rapid tissue excision, enabling faster fixation.

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Last Updated: Jun 8, 2026

A Hydrophobic Tissue Clearing Method for Rat Brain Tissue

A Hydrophobic Tissue Clearing Method for Rat Brain Tissue

Published on: December 23, 2020

Collection of Frozen Rodent Brain Regions for Downstream Analyses

Collection of Frozen Rodent Brain Regions for Downstream Analyses

Published on: April 23, 2020

Preparation of the Rat Vocal Fold for Neuromuscular Analyses

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Applying high-pressure freezing to microbiopsied tissues (brain, liver, kidney, muscle).

Employing freeze-substitution as a follow-up procedure to HPF/FS.

Main Results:

High-pressure freezing of microbiopsies effectively preserves cellular ultrastructure, minimizing artifacts.
This technique offers a non-lethal approach for obtaining certain tissue types.
Compared to chemical fixation, HPF/FS provides superior preservation of organelles and cellular morphology.

Conclusions:

Microbiopsy combined with HPF/FS is a powerful alternative to chemical fixation for electron microscopy.
This method significantly reduces fixation-induced artifacts, leading to more accurate ultrastructural analysis.
While cryosectioning offers optimal preservation, HPF/FS provides a more accessible and routine approach for high-quality sample preparation.