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Fixation and Sectioning

Fixation and Sectioning

Two basic types of preparation are used to visualize specimens with a light microscope: wet mounts and fixed specimens.
The simplest type of preparation is the wet mount, in which the specimen is placed in a drop of liquid on the slide. A liquid specimen can be directly deposited on the slide using a dropper. Solid specimens, such as skin scraping, can be placed on the slide before adding a drop of liquid to prepare the wet mount. Sometimes the liquid is simply water, but stains are often added...

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Related Experiment Video

Updated: Mar 11, 2026

Hybrid-Cut: An Improved Sectioning Method for Recalcitrant Plant Tissue Samples

Hybrid-Cut: An Improved Sectioning Method for Recalcitrant Plant Tissue Samples

Published on: November 23, 2016

Hybrid-Cut: An Improved Sectioning Method for Recalcitrant Plant Tissue Samples.

Tien-Kuan Chen¹, Hui-Ting Yang¹, Su-Chiung Fang¹

¹Academia Sinica Biotechnology Center in Southern Taiwan; Agricultural Biotechnology Research Center, Academia Sinica.

Journal of Visualized Experiments : Jove

|December 3, 2016

Summary

This study introduces the Hybrid-Cut method for plant tissue sectioning. This technique improves section integrity, especially for challenging plant tissues, enabling detailed anatomical and gene expression studies.

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Last Updated: Mar 11, 2026

Hybrid-Cut: An Improved Sectioning Method for Recalcitrant Plant Tissue Samples

Hybrid-Cut: An Improved Sectioning Method for Recalcitrant Plant Tissue Samples

Published on: November 23, 2016

In Situ Hybridization for the Precise Localization of Transcripts in Plants

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The Cutting and Floating Method for Paraffin-embedded Tissue for Sectioning

The Cutting and Floating Method for Paraffin-embedded Tissue for Sectioning

Published on: September 5, 2018

Area of Science:

Plant Anatomy
Microtechnique
Molecular Biology

Background:

Plant tissue section integrity is crucial for detailed anatomical studies.
Rigid cell walls, fibers, crystals, and high water content in plant cells challenge tissue sectioning.
Existing methods often fail to preserve delicate structures in recalcitrant plant tissues.

Purpose of the Study:

To establish a simple and effective method for preserving plant tissue integrity during sectioning.
To improve the quality of plant tissue sections for anatomical and molecular analyses.
To provide a protocol suitable for recalcitrant plant tissues.

Main Methods:

A modified paraffin-based sectioning technique, termed Hybrid-Cut, was developed.
Plant tissues were embedded in paraffin and sectioned at low temperatures (-16 °C) using a cryostat.
The low-temperature sectioning hardened paraffin blocks, minimizing tearing and scratching.

Main Results:

The Hybrid-Cut method significantly improved tissue section integrity across various plant species.
Successfully applied to calcium oxalate-rich orchid tissues and recalcitrant rice, maize, and wheat tissues.
High-quality sections were obtained, suitable for in situ hybridization (ISH) to visualize gene expression patterns.

Conclusions:

The Hybrid-Cut protocol is an effective solution for sectioning recalcitrant plant tissues, particularly those with high crystal or silica content.
This method enhances the quality of plant tissue sections, facilitating detailed morphological and biological investigations.
Improved section quality supports advanced techniques like in situ hybridization for gene expression studies.