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Chromatic Aberration-Corrected Tilt Series Transmission Electron Microscopy of Nanoparticles in a Whole Mount Macrophage Cell

Published online by Cambridge University Press:  09 May 2013

Jean-Pierre Baudoin ,
Joerg R. Jinschek ,
Chris B. Boothroyd ,
Rafal E. Dunin-Borkowski  and
Niels de Jonge
Jean-Pierre Baudoin
Affiliation:
Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, Nashville, TN 37232-0615, USA
Joerg R. Jinschek
Affiliation:
FEI Company Europe, 5600 KA Eindhoven, The Netherlands
Chris B. Boothroyd
Affiliation:
Forschungszentrum Jülich, Ernst Ruska-Centre for Microscopy and Spectroscopy with Electrons (ER-C) and Peter Grünberg Institute (PGI), D-52425 Jülich, Germany
Rafal E. Dunin-Borkowski
Affiliation:
Forschungszentrum Jülich, Ernst Ruska-Centre for Microscopy and Spectroscopy with Electrons (ER-C) and Peter Grünberg Institute (PGI), D-52425 Jülich, Germany
Niels de Jonge*
Affiliation:
Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, Nashville, TN 37232-0615, USA INM-Leibniz Institute for New Materials, D-66123 Saarbrücken, Germany
*
*Corresponding author. E-mail: [email protected]
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Abstract

Transmission electron microscopy (TEM) in combination with electron tomography is widely used to obtain nanometer scale three-dimensional (3D) structural information about biological samples. However, studies of whole eukaryotic cells are limited in resolution and/or contrast on account of the effect of chromatic aberration of the TEM objective lens on electrons that have been scattered inelastically in the specimen. As a result, 3D information is usually obtained from sections and not from whole cells. Here, we use chromatic aberration-corrected TEM to record bright-field TEM images of nanoparticles in a whole mount macrophage cell. Tilt series of images are used to generate electron tomograms, which are analyzed to assess the spatial resolution that can be achieved for different vertical positions in the specimen. The uptake of gold nanoparticles coated with low-density lipoprotein (LDL) is studied. The LDL is found to assemble in clusters. The clusters contain nanoparticles taken up on different days, which are joined without mixing their nanoparticle cargo.

Keywords

whole eukaryotic celllow-density lipoproteinspecific labelsgold nanoparticlestomographythree dimensionsspatial resolutionthick sample
Type
Biological Applications
Information
Microscopy and Microanalysis , Volume 19 , Issue 4 , August 2013 , pp. 814 - 820
Copyright
Copyright © Microscopy Society of America 2013 

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