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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
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: niels.de.jonge@inm-gmbh.de
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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.

Type
Biological Applications
Copyright
Copyright © Microscopy Society of America 2013 

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References

Aoyama, K., Takagi, T., Hirase, A. & Miyazawa, A. (2008). STEM tomography for thick biological specimens. Ultramicroscopy 109, 7080.CrossRefGoogle ScholarPubMed
Baudoin, J.P., Jerome, W.G., Kubel, C. & de Jonge, N. (2013). Whole-cell analysis of low-density lipoprotein uptake by macrophages using STEM tomography. PLoS One 8, e55022. Google Scholar
de Jonge, N., Poirier-Demers, N., Demers, H., Peckys, D.B. & Drouin, D. (2010a). Nanometer-resolution electron microscopy through micrometers-thick water layers. Ultramicroscopy 110, 11141119.CrossRefGoogle ScholarPubMed
de Jonge, N., Sougrat, R., Northan, B.M. & Pennycook, S.J. (2010b). Three-dimensional scanning transmission electron microscopy of biological specimens. Microsc Microanal 16, 5463.CrossRefGoogle ScholarPubMed
Dukes, M.J., Ramachandra, R., Baudoin, J.P., Jerome, W.G. & de Jonge, N. (2011). Three-dimensional locations of gold-labeled proteins in a whole mount eukaryotic cell obtained with 3 nm precision using aberration-corrected scanning transmission electron microscopy. J Struct Biol 174, 552562.Google Scholar
Engel, A. (2009). Scanning transmission electron microscopy: Biological applications. Adv Imag Elect Phys 159, 357386.Google Scholar
Frens, G. (1973). Controlled nucleation for regulation of particle-size in monodisperse gold suspensions. Nature Phys Sci 241, 2022.CrossRefGoogle Scholar
Gan, L. & Jensen, G.J. (2012). Electron tomography of cells. Q Rev Biophys 45, 2756.CrossRefGoogle ScholarPubMed
Gusnard, D. & Kirschner, R.H. (1977). Cell and organelle shrinkage during preparation for scanning electron microscopy: Effects of fixation, dehydration and critical point drying. J Microsc 110, 5157.Google Scholar
Haider, M., Hartel, P., Muller, H., Uhlemann, S. & Zach, J. (2009). Current and future aberration correctors for the improvement of resolution in electron microscopy. Philos Transact A Math Phys Eng Sci 367, 36653682.Google ScholarPubMed
Heymann, J.A., Hayles, M., Gestmann, I., Giannuzzi, L.A., Lich, B. & Subramaniam, S. (2006). Site-specific 3D imaging of cells and tissues with a dual beam microscope. J Struct Biol 155, 6373.Google Scholar
Hoenger, A. & McIntosh, J.R. (2009). Probing the macromolecular organization of cells by electron tomography. Curr Opin Cell Biol 21, 8996.Google Scholar
Hohmann-Marriott, M.F., Sousa, A.A., Azari, A.A., Glushakova, S., Zhang, G., Zimmerberg, J. & Leapman, R.D. (2009). Nanoscale 3D cellular imaging by axial scanning transmission electron tomography. Nat Methods 6, 729731.Google Scholar
Hohn, K., Sailer, M., Wang, L., Lorenz, M., Schneider, M.E. & Walther, P. (2011). Preparation of cryofixed cells for improved 3D ultrastructure with scanning transmission electron tomography. Histochem Cell Biol 135, 19.Google Scholar
Ikonen, E. (2008). Cellular cholesterol trafficking and compartmentalization. Nat Rev Mol Cell Biol 9, 125138.Google Scholar
Jerome, W.G., Cox, B.E., Griffin, E.E. & Ullery, J.C. (2008). Lysosomal cholesterol accumulation inhibits subsequent hydrolysis of lipoprotein cholesteryl ester. Microsc Microanal 14, 138149.Google Scholar
Jerome, W.G. & Yancey, P.G. (2003). The role of microscopy in understanding atherosclerotic lysosomal lipid metabolism. Microsc Microanal 9, 5467.CrossRefGoogle ScholarPubMed
Kabius, B., Hartel, P., Haider, M., Muller, H., Uhlemann, S., Loebau, U., Zach, J. & Rose, H. (2009). First application of Cc-corrected imaging for high-resolution and energy-filtered TEM. J Electron Microsc (Tokyo) 58, 147155.Google Scholar
Koster, A.J., Grimm, R., Typke, D., Hegerl, R., Stoschek, A., Walz, J. & Baumeister, W. (1997). Perspectives of molecular and cellular electron tomography. J Struct Biol 120, 276308.Google Scholar
Kourkoutis, L.F., Plitzko, J.M. & Baumeister, W. (2012). Electron microscopy of biological materials at the nanometer scale. Annu Rev Mater Res 42, 3358.Google Scholar
Kremer, J.R., Mastronarde, D.N. & McIntosch, J.R. (1996). Computer visualization of three-dimensional image data using IMOD. J Struct Biol 116, 7176.Google Scholar
Larabell, C.A. & Nugent, K.A. (2010). Imaging cellular architecture with X-rays. Curr Opin Struct Biol 20, 623631.Google Scholar
Leary, R. & Brydson, R. (2011). Chromatic aberration correction: The next step on electron microscopy. Adv Imag Elect Phys 165, 73130.Google Scholar
Luzio, J.P., Pryor, P.R. & Bright, N.A. (2007). Lysosomes: Fusion and function. Nat Rev Mol Cell Biol 8, 622632.Google Scholar
Medalia, O., Weber, I., Frangakis, A.S., Nicastro, D., Gerisch, G. & Baumeister, W. (2002). Macromolecular architecture in eukaryotic cells visualized by cryoelectron tomography. Science 298, 12091213.CrossRefGoogle ScholarPubMed
Moore, K.J. & Tabas, I. (2011). Macrophages in the pathogenesis of atherosclerosis. Cell 145, 341355.Google Scholar
Pierson, J., Sani, M., Tomova, C., Godsave, S. & Peters, P.J. (2009). Toward visualization of nanomachines in their native cellular environment. Histochem Cell Biol 132, 253262.Google Scholar
Radermacher, M.H.W. (1980). Properties of three-dimensionally reconstructed objects from projections by conical tilting compared to single axis tilting. Proc 7th Eur Congr Electron Microsc Den Haag 1, 132133.Google Scholar
Reimer, L. & Gentsch, P. (1975). Superposition of chromatic error and beam broadening in transmission electron microscopy of thick carbon and organic specimens. Ultramicroscopy 1, 15.Google Scholar
Reimer, L. & Kohl, H. (2008). Transmission Electron Microscopy: Physics of Image Formation. New York: Springer.Google Scholar
Reimer, L. & Ross-Messemer, M. (1987). Top-bottom effect in energy-selecting transmission electron microscopy. Ultramicroscopy 21, 385387.Google Scholar
Sousa, A.A., Aronova, M.A., Kim, Y.C., Dorward, L.M., Zhang, G. & Leapman, R.D. (2007). On the feasibility of visualizing ultrasmall gold labels in biological specimens by STEM tomography. J Struct Biol 159, 507522.CrossRefGoogle ScholarPubMed
van Aert, S., Batenburg, K.J., Rossell, M.D., Erni, R. & van Tendeloo, G. (2011). Three-dimensional atomic imaging of crystalline nanoparticles. Nature 470, 374377.Google Scholar
Yakushevska, A.E., Lebbink, M.N., Geerts, W.J., Spek, L., van Donselaar, E.G., Jansen, K.A., Humbel, B.M., Post, J.A., Verkleij, A.J. & Koster, A.J. (2007). STEM tomography in cell biology. J Struct Biol 159, 381391.Google Scholar
Yancey, P.G., Miles, S., Schwegel, J. & Jerome, W.G. (2002). Uptake and trafficking of mildly oxidized LDL and acetylated LDL in THP-1 cells does not explain the differences in lysosomal metabolism of these two lipoproteins. Microsc Microanal 8, 8193.Google Scholar

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