Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

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...

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...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

Global survey of microscopy core facilities.

Journal of microscopy·2026

Same author

Global survey of Flow Cytometry core facilities 2021.

Journal of microscopy·2026

Same author

Contextual High-Throughput 3D Volume Electron Microscopy Data Acquisition Using Artificial Intelligence.

Microscopy and microanalysis : the official journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada·2026

Same author

NCR13 peptide protects soybean against Cercospora sojina using multi-faceted modes of action and additive interaction with chemical fungicides.

Journal of experimental botany·2025

Same author

Best practices in plant fluorescence imaging and reporting: A primer.

The Plant cell·2025

Same author

Plants in the expansion era: recent advances in expansion microscopy in plant systems.

Journal of experimental botany·2025

Same journal

Quantification of cell viability by automated analysis of live cell imaging.

Methods in cell biology·2026

Same journal

Flow cytometry evaluation of cytotoxicity exerted by effector immune cells against tumor cells.

Methods in cell biology·2026

Same journal

Time-lapse confocal laser scanning microscopy analysis of FOOD formation.

Methods in cell biology·2026

Same journal

Screening and identification of protein-protein interaction using proximity labeling.

Methods in cell biology·2026

Same journal

Quantitative high-content profiling of mitochondrial morphology with automated statistical analysis and integrated data visualization.

Methods in cell biology·2026

Same journal

Super-resolution imaging of cell death in Drosophila tissues via expansion and pan-expansion microscopy.

Methods in cell biology·2026

See all related articles

Search research articles

Related Experiment Video

Updated: Jul 23, 2025

Microscopy Techniques for Interpreting Fungal Colonization in Mycoheterotrophic Plants Tissues and Symbiotic Germination of Seeds

Microscopy Techniques for Interpreting Fungal Colonization in Mycoheterotrophic Plants Tissues and Symbiotic Germination of Seeds

Published on: May 17, 2022

A conventional fixation volume electron microscopy protocol for plants.

Janithri S Wickramanayake¹, Kirk J Czymmek¹

¹Donald Danforth Plant Science Center, Saint Louis, MO, United States; Advanced Bioimaging Laboratory, Donald Danforth Plant Science Center, Saint Louis, MO, United States.

Methods in Cell Biology

|July 14, 2023

Summary

This summary is machine-generated.

This study presents a new chemical fixation protocol for plant tissues, enabling high-resolution volume electron microscopy (EM) of cellular structures. The optimized method overcomes challenges posed by plant sample properties for advanced EM imaging.

Keywords:

Chloroplasts Duckweed Focused ion-beam scanning electron microscopy Nicotiana benthamiana OTO Osmium-thiocarbohydrazide-osmium Serial block-face scanning electron microscopy Tobacco

More Related Videos

Scanning Electron Microscopy SEM Protocols for Problematic Plant, Oomycete, and Fungal Samples

Scanning Electron Microscopy SEM Protocols for Problematic Plant, Oomycete, and Fungal Samples

Published on: February 3, 2017

Tandem High-pressure Freezing and Quick Freeze Substitution of Plant Tissues for Transmission Electron Microscopy

Tandem High-pressure Freezing and Quick Freeze Substitution of Plant Tissues for Transmission Electron Microscopy

Published on: October 13, 2014

Related Experiment Videos

Last Updated: Jul 23, 2025

Microscopy Techniques for Interpreting Fungal Colonization in Mycoheterotrophic Plants Tissues and Symbiotic Germination of Seeds

Microscopy Techniques for Interpreting Fungal Colonization in Mycoheterotrophic Plants Tissues and Symbiotic Germination of Seeds

Published on: May 17, 2022

Scanning Electron Microscopy SEM Protocols for Problematic Plant, Oomycete, and Fungal Samples

Scanning Electron Microscopy SEM Protocols for Problematic Plant, Oomycete, and Fungal Samples

Published on: February 3, 2017

Tandem High-pressure Freezing and Quick Freeze Substitution of Plant Tissues for Transmission Electron Microscopy

Tandem High-pressure Freezing and Quick Freeze Substitution of Plant Tissues for Transmission Electron Microscopy

Published on: October 13, 2014

Area of Science:

Plant Biology
Microscopy
Cell Biology

Background:

Volume electron microscopy (EM) is crucial for plant research, but plant tissues present unique challenges for high-resolution imaging.
Challenges include large air voids, cell walls, and waxy cuticles, hindering morphology, staining, and conductivity.

Purpose of the Study:

To develop and validate a robust chemical fixation protocol for preparing plant samples for high-resolution volume EM.
To address the specific challenges associated with plant tissues for techniques like serial block-face scanning electron microscopy (SB-SEM) and focused ion beam scanning electron microscopy (FIB-SEM).

Main Methods:

A modified chemical fixation strategy was employed, starting with paraformaldehyde and glutaraldehyde, followed by secondary fixation with osmium tetroxide, potassium ferrocyanide, thiocarbohydrazide, and osmium tetroxide.
Further staining involved uranyl acetate and lead aspartate, dehydration in acetone, and embedding in Quetol 651 resin.
Samples were imaged using SB-SEM with focal charge compensation.

Main Results:

The protocol successfully produced high-contrast images of cellular structures in tobacco and duckweed leaf tissues.
Key organelles such as mitochondria, Golgi, endoplasmic reticulum, and nuclear envelope membranes were clearly visualized.
Chloroplast thylakoid membranes and grana lamellae were distinctly resolved, demonstrating the protocol's efficacy.

Conclusions:

This optimized sample preparation protocol provides a reliable method for routine plant volume electron microscopy.
It effectively overcomes common challenges in plant tissue preparation for advanced EM techniques.
The protocol facilitates detailed ultrastructural analysis of plant cells and tissues.