Skip to main content Accessibility help
×
Home

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

  • Jean-Pierre Baudoin (a1), Joerg R. Jinschek (a2), Chris B. Boothroyd (a3), Rafal E. Dunin-Borkowski (a3) and Niels de Jonge (a1) (a4)...

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.

Copyright

Corresponding author

* Corresponding author. E-mail: niels.de.jonge@inm-gmbh.de

References

Hide All
Aoyama, K., Takagi, T., Hirase, A. & Miyazawa, A. (2008). STEM tomography for thick biological specimens. Ultramicroscopy 109, 7080.
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.
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.
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.
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.
Engel, A. (2009). Scanning transmission electron microscopy: Biological applications. Adv Imag Elect Phys 159, 357386.
Frens, G. (1973). Controlled nucleation for regulation of particle-size in monodisperse gold suspensions. Nature Phys Sci 241, 2022.
Gan, L. & Jensen, G.J. (2012). Electron tomography of cells. Q Rev Biophys 45, 2756.
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.
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.
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.
Hoenger, A. & McIntosh, J.R. (2009). Probing the macromolecular organization of cells by electron tomography. Curr Opin Cell Biol 21, 8996.
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.
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.
Ikonen, E. (2008). Cellular cholesterol trafficking and compartmentalization. Nat Rev Mol Cell Biol 9, 125138.
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.
Jerome, W.G. & Yancey, P.G. (2003). The role of microscopy in understanding atherosclerotic lysosomal lipid metabolism. Microsc Microanal 9, 5467.
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.
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.
Kourkoutis, L.F., Plitzko, J.M. & Baumeister, W. (2012). Electron microscopy of biological materials at the nanometer scale. Annu Rev Mater Res 42, 3358.
Kremer, J.R., Mastronarde, D.N. & McIntosch, J.R. (1996). Computer visualization of three-dimensional image data using IMOD. J Struct Biol 116, 7176.
Larabell, C.A. & Nugent, K.A. (2010). Imaging cellular architecture with X-rays. Curr Opin Struct Biol 20, 623631.
Leary, R. & Brydson, R. (2011). Chromatic aberration correction: The next step on electron microscopy. Adv Imag Elect Phys 165, 73130.
Luzio, J.P., Pryor, P.R. & Bright, N.A. (2007). Lysosomes: Fusion and function. Nat Rev Mol Cell Biol 8, 622632.
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.
Moore, K.J. & Tabas, I. (2011). Macrophages in the pathogenesis of atherosclerosis. Cell 145, 341355.
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.
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.
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.
Reimer, L. & Kohl, H. (2008). Transmission Electron Microscopy: Physics of Image Formation. New York: Springer.
Reimer, L. & Ross-Messemer, M. (1987). Top-bottom effect in energy-selecting transmission electron microscopy. Ultramicroscopy 21, 385387.
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.
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.
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.
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.

Keywords

Type Description Title
VIDEO
Supplementary materials

Baudoin Supplementary Material
Movie 1

 Video (930 KB)
930 KB
VIDEO
Supplementary materials

Baudoin Supplementary Material
Movie 2

 Video (11.4 MB)
11.4 MB
VIDEO
Supplementary materials

Baudoin Supplementary Material
Movie 3

 Video (1.6 MB)
1.6 MB
VIDEO
Supplementary materials

Baudoin Supplementary Material
Movie 4

 Video (4.0 MB)
4.0 MB
VIDEO
Supplementary materials

Baudoin Supplementary Material
Movie 5

 Video (14.2 MB)
14.2 MB
PDF
Supplementary materials

Baudoin Supplementary Material
Movie Legends

 PDF (239 KB)
239 KB

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

  • Jean-Pierre Baudoin (a1), Joerg R. Jinschek (a2), Chris B. Boothroyd (a3), Rafal E. Dunin-Borkowski (a3) and Niels de Jonge (a1) (a4)...

Metrics

Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed