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Overview of Electron Microscopy

Overview of Electron Microscopy

The wavelengths of visible light ultimately limit the maximum theoretical resolution of images created by light microscopes. Most light microscopes can only magnify 1000X, and a few can magnify up to 1500X. Electrons, like electromagnetic radiation, can behave like waves, but with wavelengths of 0.005 nm, they produce significantly greater resolution up to 0.05 nm as compared to 500 nm for visible light. An electron microscope (EM) can create a sharp image that is magnified up to 2,000,000X.

Cryo-electron Microscopy

Cryo-electron Microscopy

Conventional electron microscopy (EM) involves dehydration, fixation, and staining of biological samples, which distorts the native state of biological molecules and results in several artifacts. Also, the high-energy electron beam damages the sample and makes it difficult to obtain high-resolution images. These issues can be addressed using cryo-EM, which uses frozen samples and gentler electron beams. The technique was developed by Jacques Dubochet, Joachim Frank, and Richard Henderson, for...

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Electron microscopy in yeast.

¹Department of Chemical and Biological Sciences, Faculty of Science, Japan Women's University, Tokyo, Japan.

Methods in Enzymology

|February 3, 2009

Summary

Electron microscopy and cryofixation reveal yeast membrane dynamics during autophagy and the cytoplasm-to-vacuole targeting (Cvt) pathway. These methods clarify similarities between these essential cellular processes.

Area of Science:

Cell Biology
Molecular Biology

Background:

Autophagy is a crucial cellular process for degrading and recycling cellular components.
The cytoplasm-to-vacuole targeting (Cvt) pathway shares mechanistic similarities with autophagy.
Understanding the membrane dynamics of these pathways is key to cellular health.

Purpose of the Study:

To describe the application of electron microscopy (EM) methods for studying membrane dynamics in yeast.
To illustrate the dynamic events in autophagy and the Cvt pathway using EM.
To highlight the similarities between autophagy and the Cvt pathway.

Main Methods:

Cryofixation technique for rapid sample preservation.
Various electron microscopy (EM) methods, including transmission electron microscopy (TEM).

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High-resolution imaging of cellular structures and dynamic events.

Main Results:

Detailed visualization of membrane dynamics during yeast autophagy.
Clarification of the dynamic process of the Cvt pathway.
Direct comparison of EM images revealing similarities between autophagy and Cvt pathway membrane dynamics.

Conclusions:

Electron microscopy combined with cryofixation is effective for resolving yeast membrane dynamics.
EM studies have elucidated the similarities between the autophagy and Cvt pathways.
This chapter provides a visual guide to these essential cellular processes in yeast.