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Related Experiment Video

Updated: May 18, 2026

Mitochondria and Endoplasmic Reticulum Imaging by Correlative Light and Volume Electron Microscopy

Mitochondria and Endoplasmic Reticulum Imaging by Correlative Light and Volume Electron Microscopy

Published on: July 20, 2019

Electron microscopy of endocytic pathways.

Carmen Ranftler¹, Peter Auinger, Claudia Meisslitzer-Ruppitsch

¹Department of Cell Biology and Ultrastructure Research, Center for Anatomy and Cell Biology, Medical University of Vienna, Vienna, Austria.

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

|October 3, 2012

Summary

This summary is machine-generated.

This study introduces a new cytochemical method using wheat germ agglutinin (WGA) for electron microscopy. This technique enhances visualization of endocytic pathways and cellular transport, improving ultrastructural analysis.

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Area of Science:

Cell Biology
Microscopy
Biochemistry

Background:

Understanding the endocytic system's architecture is crucial for analyzing retrograde transport.
Existing methods for visualizing endocytic pathways have limitations in resolution and preservation.

Purpose of the Study:

To develop and validate a cytochemical approach for detailed electron microscopic study of endocytic pathways.
To combine wheat germ agglutinin (WGA) labeling with advanced fixation techniques for enhanced ultrastructural analysis.

Main Methods:

Utilized horseradish peroxidase-labeled WGA as a marker for endocytosis in human hepatoma cells.
Employed both conventional chemical fixation and high pressure-freezing (HPF) for cell fixation.
Visualized intracellular routes via diaminobenzidine (DAB) oxidation, performed before or after fixation.

Main Results:

The combined cytochemistry and HPF protocol yielded distinct cytochemical reactions.
Achieved excellently preserved fine structures suitable for detailed electron microscopy and 3D reconstruction.
Demonstrated the technique's efficacy in studying complex and dynamic endocytic architectures.

Conclusions:

The described cytochemical approach, particularly with HPF, offers high spatial and temporal resolution for endocytic pathway analysis.
This method provides superior ultrastructural detail for morpho-functional studies of intracellular transport.