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Liquid Scanning Transmission Electron Microscopy: Imaging Protein Complexes in their Native Environment in Whole Eukaryotic Cells

Published online by Cambridge University Press:  19 February 2014

Diana B. Peckys  and
Niels de Jonge
Diana B. Peckys
Affiliation:
Leibniz Institute for New Materials (INM), 66123 Saarbrücken, Germany
Niels de Jonge*
Affiliation:
Leibniz Institute for New Materials (INM), 66123 Saarbrücken, Germany Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, Nashville, TN 37232-0615, USA
*
*Corresponding author. [email protected]
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Abstract

Scanning transmission electron microscopy (STEM) of specimens in liquid, so-called Liquid STEM, is capable of imaging the individual subunits of macromolecular complexes in whole eukaryotic cells in liquid. This paper discusses this new microscopy modality within the context of state-of-the-art microscopy of cells. The principle of operation and equations for the resolution are described. The obtained images are different from those acquired with standard transmission electron microscopy showing the cellular ultrastructure. Instead, contrast is obtained on specific labels. Images can be recorded in two ways, either via STEM at 200 keV electron beam energy using a microfluidic chamber enclosing the cells, or via environmental scanning electron microscopy at 30 keV of cells in a wet environment. The first series of experiments involved the epidermal growth factor receptor labeled with gold nanoparticles. The labels were imaged in whole fixed cells with nanometer resolution. Since the cells can be kept alive in the microfluidic chamber, it is also feasible to detect the labels in unfixed, live cells. The rapid sample preparation and imaging allows studies of multiple whole cells.

Keywords

Liquid STEMESEM-STEMgold nanoparticlespecific labelepidermal growth factor receptor (EGFR)whole celllive cell microscopyenvironmental scanning electron microscopy
Type
In Situ Special Section
Information
Microscopy and Microanalysis , Volume 20 , Issue 2 , April 2014 , pp. 346 - 365
Copyright
© Microscopy Society of America 2014 

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