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Variations on Negative Stain Electron Microscopy Methods: Tools for Tackling Challenging Systems

Variations on Negative Stain Electron Microscopy Methods: Tools for Tackling Challenging Systems

Published on: February 6, 2018

A negative stain for electron microscopic tomography.

Andrea Fera¹, Jane E Farrington, Joshua Zimmerberg

¹Laboratory of Neurobiology, National Institute of Neurological Disorders and Stroke, Bethesda, MD 20892, USA.

Microscopy and Microanalysis : the Official Journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada

|February 28, 2012

Summary

This summary is machine-generated.

Negative stain tomography offers a new method for 3D imaging of biological structures. This technique, using a tungsten-based stain, provides detailed views of individual proteins on the influenza A virus surface.

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Area of Science:

Structural Biology
Electron Microscopy

Background:

Negative staining provides 2D images of biological structures.
3D imaging via negative staining and tomography is underdeveloped.

Purpose of the Study:

To evaluate a tungsten-based stain for negative stain tomography.
To assess the feasibility of 3D imaging of biological structures using this method.

Main Methods:

Utilized a commercially available tungsten-based stain.
Collected tomographic images of negatively stained influenza A virus.
Compared resulting surface renderings with cryomicroscopy and crystallized protein data.

Main Results:

The tungsten-based stain met prerequisites for high-dose electron microscopy.
Detailed 3D images of individual viral surface proteins were obtained.
Surface renderings closely matched existing cryomicroscopy and crystallographic data.

Conclusions:

Negative stain tomography with an appropriate stain is a viable method for 3D structural analysis.
This technique allows visualization of individual molecules in their native context.
It offers a valuable approach for studying viral surface structures.