Abstract
The electron microscope has made paramount contributions to understanding the structure of biological molecules, cells, and tissues. In general, the most faithful preservation of biological specimens and other soft-organic materials is achieved through cryogenic fixation. The embedding medium is the native aqueous environment itself, immobilized in vitrified form by rapid or pressurized cooling. Until recently, imaging of such vitrified thin specimens by electron cryo-microscopy has been nearly synonymous with wide-field transmission electron microscopy (TEM). Several new approaches have entered the cryo-microscopy field, including soft x-ray imaging, serial surface imaging using focused ion beam scanning electron microscopy, phase plates, and scanning TEM (STEM). In this article, we focus on the STEM method and its adaptation to biological cryo-microscopy. Cryogenic imaging of unstained specimens by STEM introduces specific challenges. Difficulties were long considered insurmountable, and the potential advantages were underappreciated. Future developments in experimental setup and detector technologies will allow for extension of the method to thicker specimens with improved resolution and analytic capabilities.
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References
J. Frank, Electron Tomography: Methods for Three-Dimensional Visualization of Structures in the Cell (Springer, New York, 2008).
L. Gan, G.J. Jensen, Q. Rev. Biophys. 45, 27 (2012).
K.D. Micheva, S.J. Smith, Neuron 55, 25 (2007).
I. Wacker, R. Schroeder, J. Microsc. 252, 93 (2013).
W. Denk, H. Horstmann, PLoS Biol. 2, e329 (2004).
J.A.W. Heymann, D. Shi, S. Kim, D. Bliss, J.L.S. Milne, S. Subramaniam, J. Struct. Biol. 166, 1 (2009).
A. Weiner, N. Dahan-Pasternak, E. Shimoni, V. Shinder, P. von Huth, M. Elbaum, R. Dzikowski, Cell. Microbiol. 13, 967 (2011).
D.J. De Rosier, A. Klug, Nature 217, 130 (1968).
X. Shu, V. Lev-Ram, T.J. Deerinck, Y. Qi, E.B. Ramko, M.W. Davidson, Y. Jin, M.H. Ellisman, R.Y. Tsien, PLoS Biol. 9, e1001041 (2011).
E. Diestra, J. Fontana, P. Guichard, S. Marco, C. Risco, J. Struct. Biol. 165, 157 (2009).
S. Watanabe, A. Punge, G. Hollopeter, K.I. Willig, R.J. Hobson, M.W. Davis, S.W. Hell, E.M. Jorgensen, Nat. Methods 8, 80 (2011).
P. Zhang, Curr. Opin. Struct. Biol. 23, 763 (2013).
N. Mellouk, A. Weiner, N. Aulner, C. Schmitt, M. Elbaum, S.L. Shorte, A. Danckaert, J. Enninga, Cell Host Microbe 16, 517 (2014).
J. Koster, K. Grünewald, Ultramicroscopy 143, 1 (2014).
H. Falls, H.T. Davis, L.E. Scriven, Y. Talmon, Biochim. Biophys. Acta Biomembr. 693, 364 (1982).
J. Lepault, F.P. Booy, J. Dubochet, J. Microsc. 129, 89 (1983).
R. Dahl, L.A. Staehelin, J. Electron Microsc. Tech. 13, 165 (1989).
O. Medalia, I. Weber, A.S. Frangakis, D. Nicastro, G. Gerisch, W. Baumeister, Science 298, 1209 (2002).
V. Lučič, A. Rigort, W. Baumeister, J. Cell Biol. 202, 407 (2013).
A. Diebolder, A.J. Koster, R.I. Koning, J. Microsc. 248, 1 (2012).
W. Kühlbrandt, eLife 3, e03678 (2014).
A. Al-Amoudi, J.-J. Chang, A. Leforestier, A. McDowall, L.M. Salamin, L.P. Norlén, K. Richter, N.S. Blanc, D. Studer, J. Dubochet, EMBO J. 23, 3583 (2004).
E. Villa, M. Schaffer, J.M. Plitzko, W. Baumeister, Curr. Opin. Struct. Biol. 23, 771 (2013).
A. Hsieh, T. Schmelzer, G. Kishchenko, T. Wagenknecht, M. Marko, J. Struct. Biol. 185, 32 (2014).
X. Li, P. Mooney, S. Zheng, C.R. Booth, M.B. Braunfeld, S. Gubbens, D.A. Agard, Y. Cheng, Nat. Methods 10, 584 (2013).
T.A.M. Bharat, C.J. Russo, J. Löwe, L.A. Passmore, S.H.W. Scheres, Structure 23, 1743 (2015).
G. Schneider, P. Guttmann, S. Heim, S. Rehbein, F. Mueller, K. Nagashima, J.B. Heymann, W.G. Müller, J.G. McNally, Nat. Methods 7, 985 (2010).
S. Kapishnikov, A. Weiner, E. Shimoni, P. Guttmann, G. Schneider, N. Dahan-Pasternak, R. Dzikowski, L. Leiserowitz, M. Elbaum, Proc. Natl. Acad. Sci. U.S.A. 109, 11188 (2012).
M. Do, S.A. Isaacson, G. McDermott, M.A. Le Gros, C.A. Larabell, Arch. Biochem. Biophys. (2015), doi:10.1016/j.abb.2015.01.011.
A. Schertel, N. Snaidero, H.-M. Han, T. Ruhwedel, M. Laue, M. Grabenbauer, W. Möbius, J. Struct. Biol. 184, 355 (2013).
N. Vidavsky, A. Masic, A. Schertel, S. Weiner, L. Addadi, J. Struct. Biol. 192, 358 (2015).
S.G. Wolf, L. Houben, M. Elbaum, Nat. Methods 11, 423 (2014).
A. Muller, Nat. Mater. 8, 263 (2009).
O.L. Krivanek, M.F. Chisholm, V. Nicolosi, T.J. Pennycook, G.J. Corbin, N. Dellby, M.F. Murfitt, C.S. Own, Z.S. Szilagyi, M.P. Oxley, S.T. Pantelides, S.J. Pennycook, Nature 464, 571 (2010).
S.J. Pennycook, Ultramicroscopy 123, 28 (2012).
C. Colliex, C. R. Phys. 15, 101 (2014).
C. Colliex, A. Gloter, K. March, C. Mory, O. Stéphan, K. Suenaga, M. Tencé, Ultramicroscopy 123, 80 (2012).
J.S. Wall, J.F. Hainfeld, Annu. Rev. Biophys. Biophys. Chem. 15, 355 (1986).
A. Engel, in Advances in Imaging and Electron Physics, P.W. Hawkes, Ed. (Elsevier, San Diego, 2009), vol. 159, pp. 357–386.
S. Trachtenberg, K.R. Leonard, W. Tichelaar, Ultramicroscopy 45, 307 (1992).
E. Carlemalm, E. Kellenberger, EMBO J. 1, 63 (1982).
E. Kellenberger, E. Carlemalm, W. Villiger, M. Wurtz, C. Mory, C. Colliex, Ann. N.Y. Acad. Sci. 483, 202 (1986).
J.S. Wu, A.M. Kim, R. Bleher, B.D. Myers, R.G. Marvin, H. Inada, K. Nakamura, X.F. Zhang, E. Roth, S.Y. Li, T.K. Woodruff, T.V. O’Halloran, V.P. Dravid, Ultramicroscopy 128, 24 (2013).
T. Klein, E. Buhr, C.G. Frase, in Advances in Imaging and Electron Physics, P.W. Hawkes, Ed. (Elsevier, San Diego, 2012), vol. 171, pp. 297–356.
M.A. Aronova, R.D. Leapman, MRS Bull. 37, 53 (2012).
A.E. Yakushevska, M.N. Lebbink, W.J.C. Geerts, L. Spek, E.G. van Donselaar, K.A. Jansen, B.M. Humbel, J.A. Post, A.J. Verkleij, A.J. Koster, J. Struct. Biol. 159, 381 (2007).
K. Aoyama, T. Takagi, A. Hirase, A. Miyazawa, Ultramicroscopy 109, 70 (2008).
M.F. Hohmann-Marriott, A.A. Sousa, A.A. Azari, S. Glushakova, G. Zhang, J. Zimmerberg, R.D. Leapman, Nat. Methods 6, 729 (2009).
D. Charuvi, V. Kiss, R. Nevo, E. Shimoni, Z. Adam, Z. Reich, Plant Cell 24, 1143–1157 (2012).
Y. Mutsafi, E. Shimoni, A. Shimon, A. Minsky, PLoS Pathog. 9, e1003367 (2013).
O. Tsabari, R. Nevo, S. Meir, L.R. Carrillo, D.M. Kramer, Z. Reich, Plant J. 81, 884 (2015).
E. Milrot, Y. Mutsafi, Y. Fridmann-Sirkis, E. Shimoni, K. Rechav, J.R. Gurnon, J.L. Van Etten, A. Minsky, Cell. Microbiol. 18, 3 (2016).
S. Boonrungsiman, E. Gentleman, R. Carzaniga, N.D. Evans, D.W. McComb, A.E. Porter, M.M. Stevens, Proc. Natl. Acad. Sci. U.S.A. 109, 14170 (2012).
K. Murata, M. Esaki, T. Ogura, S. Arai, Y. Yamamoto, N. Tanaka, Ultramicroscopy 146, 39 (2014).
C. Suarez, G. Andres, A. Kolovou, S. Hoppe, M.L. Salas, P. Walther, J. Krijnse-Locker, Cell. Microbiol. 17, 1683 (2015).
E. Papanikou, K.J. Day, J.A. Ii, B.S. Glick, eLife 4, e13232 (2015).
I.D. Pokrovskaya, M.A. Aronova, J.A. Kamykowski, A.A. Prince, J.D. Hoyne, G.N. Calco, B.C. Kuo, Q. He, R.D. Leapman, B. Storrie, J. Thromb. Haemost. 14, 572 (2016).
D. Kirchenbuechler, Y. Mutsafi, B. Horowitz, S. Levin-Zaidman, D. Fass, S.G. Wolf, M. Elbaum, AIMS Biophys. 2, 259 (2015).
A. Philippsen, H.-A. Engel, A. Engel, Ultramicroscopy 107, 202 (2007).
A. Zanetti, J.D. Riches, S.D. Fuller, J.A.G. Briggs, J. Struct. Biol. 168, 305 (2009).
F. Zernike, Science 121, 345 (1955).
K. Nagayama, J. Electron Microsc. (Tokyo) 60 (Suppl. 1), S43 (2011).
R.M. Glaeser, Rev. Sci. Instrum. 84, 111101 (2013).
R. Danev, B. Buijsse, M. Khoshouei, J.M. Plitzko, W. Baumeister, Proc. Natl. Acad. Sci. U.S.A. 111, 15635 (2014).
J. Mahamid, S. Pfeffer, M. Schaffer, E. Villa, R. Danev, L.K. Cuellar, F. Förster, A.A. Hyman, J.M. Plitzko, W. Baumeister, Science 351, 969 (2016).
C. Ophus, J. Ciston, J. Pierce, T.R. Harvey, J. Chess, B.J. McMorran, C. Czarnik, H.H. Rose, P. Ercius, Nat. Commun. 7, 10719 (2016).
J.K. Hyun, P. Ercius, D.A. Muller, Ultramicroscopy 109, 1 (2008).
J. Biskupek, J. Leschner, P. Walther, U. Kaiser, Ultramicroscopy 110, 1231 (2010).
P. Rez, Ultramicroscopy 96, 117 (2003).
H. Rose, Ninth Int. Congr. Electron Microsc. Tor. III, 230–243 (1978).
S. Trepout, C. Messaoudi, S. Perrot, P. Bastin, S. Marco, Micron 77, 9 (2015).
L.R. Comolli, K.H. Downing, J. Struct. Biol. 152, 149 (2005).
S.G. Wolf, P. Rez, M. Elbaum, J. Microsc. 260, 227 (2015).
M. Seufferheld, M.C.F. Vieira, F.A. Ruiz, C.O. Rodrigues, S.N.J. Moreno, R. Docampo, J. Biol. Chem. 278, 29971 (2003).
F.G.A. Faas, M.C. Avramut, B.M. van den Berg, A.M. Mommaas, A.J. Koster, R.B.G. Ravelli, J. Cell Biol. 198, 457 (2012).
L. Gan, M.S. Ladinsky, G.J. Jensen, Chromosoma 122, 377 (2013).
I. Romero-Brey, R. Bartenschlager, Viruses 7, 6316 (2015).
A.V. Crewe, J. Wall, J. Langmore, Science 168, 1338 (1970).
E.J. Kirkland, in Advanced Computing in Electron Microscopy (Springer, New York), pp. 1–4.
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Elbaum, M., Wolf, S.G. & Houben, L. Cryo-scanning transmission electron tomography of biological cells. MRS Bulletin 41, 542–548 (2016). https://doi.org/10.1557/mrs.2016.136
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DOI: https://doi.org/10.1557/mrs.2016.136