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High-speed AFM R&D Division

Super-resolution AFM R&D Division

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walking myosin V

Proteins are dynamic in nature and work at the single-molecule level. Reflecting this fact, single-molecule fluorescence microscopy has been widely exploited to understand how proteins operate. However, what we can observe thereby is the dynamic behaviour of individual fluorescent spots (each being emitted from a fluorophore attached to a selected locus of the molecule), not of the protein molecules themselves. The structure of proteins has been studied by electron microscopy, X-ray crystallography and NMR, but the obtained structures are essentially static. As such, the simultaneous assessment of structure and dynamics has long been impossible. To overcome this limitation, we have been developing various techniques for increasing the imaging speed of AFM since 1993, and now a practically useful high-speed AFM is established.

Ongoing Research:  We are now developing the next generation of atomic force microscopy (AFM) techniques including a tip-scan type of high-speed AFM instrument that can be combined with various optical microscopy techniques. We are also working on the commercialization of the established high-speed AFM instrument, collaborating with manufacturers. 

Selected Publications@

T. Ando, "High-speed atomic force microscopy coming of age", Nanotechnology 23: 062001 (27 pp)  (2012).

K. Igarashi, T. Uchihashi, A. Koivula, M. Wada, S. Kimura, T. Okamoto, M. Penttilä, T. Ando, and M. Samejima, "Traffic jams reduce hydrolytic efficiency of cellulase on cellulose surface", Science 333:1279-1282 (2011).

T. Uchihashi, R. Iino, T. Ando, and H. Noji, "High-speed atomic force microscopy reveals rotory catalysis of rotorless F1-ATPase, Science 333: 755-758 (2011).

N. Kodera, D. Yamamoto, R. Ishikawa, and T. Ando, "Video imaging of walking myosin V by high-speed atomic force microscopy", Nature 468: 72-76 (2010).  (supplementary movies) (supplementary info) (News & View)

M. Shibata, T. Uchihashi, H. Yamashita, H. Kandori, and T. Ando, "Structural changes in bacteriorhodopsin in response to alternate illumination observed by high-speed atomic force microscopy",  Angewande Chemie International edition 50: 4410 –4413 (2011).

P.-E. Milhiet, D. Yamamoto., O. Berthoumieu, P. Dosset, C. Le Grimellec, J.-M.Verdier, S. Marchal, and T. Ando, "Deciphering the structure, growth and assembly of amyloid-like fibrils using high-speed atomic force microscopy", PLos One 5 (11): e13240 (8 pp) (2010).  (Movie S1, Movie S2, Movie S3) (supplementary info)

D. Yamamoto, T. Uchihashi, N. Kodera, H. Yamashita, S. Nishikori, T. Ogura, M. Shibata, and T. Ando, "High-speed atomic force microscopy techniques for observing dynamic biomolecular processes", Methods Enzymol. 475 (Part B): 541-564 (2010).

D. Yamamoto, T. Azuma, T. Uchihashi, H. Sasaki, H. Watanabe, T. Ando, and Y. Fukumori,  "Visualization and structural analysis of the bacterial magnetic organelle magnetosome using atomic force microscopy", Proc. Natl. Acad. Sci. USA 107: 9382-9387 (2010). (supplementary info) (supplementary movie)

S. Sugimoto, K. Yamanaka, S. Nishikori, A. Miyagi, T. Ando, and T. Ogura, "AAA+ chaperone ClpX regurates dynamics of prokaryotic cytoskeletal protein FtsZ", J. Biol. Chem. 285: 6648-6657 (2010).

M.-C. Giocondi, D. Yamamoto, E. Lesniewska, P.-E. Milhiet, T. Ando, and C. Le Grimellec, "Surface topography of membrane domains", Bioochim. Biophys. Acta.-Biomembranes 1798: 703-718 (2010)

M. Shibata, H. Yamashita, T. Uchihashi, H. Kandori, and T. Ando, "High-speed atomic force microscopy shows dynamic molecular processes in photo-activated bacteriorhodopsin", Nature Nanotechnology 5, 208 - 212 (2010).  (Supplementary Info) (Supplementary Movies) (npg asia materials highlight)

D. Yamamoto, N. Nagura, S. Omote, M. Taniguchi, and T. Ando, "Streptavidin 2D crystal substrates for visualizing biomolecular processes by atomic force microscopy", Biophys. J. 97(8): 2358–2367 (2009).

H. Yamashita, K. Voïtchovsky, T. Uchihashi, S. Antoranz Contera, J. F. Ryan, and T. Ando, "Dynamics of bacteriorhodopsin 2D crystal observed by high-speed atomic force microscopy", J. Struct. Biol. 167: 153-158 (2009).

http://www.sciencedirect.com/science/journal/10478477 (Supplementary Movie)

T. Ando, T. Uchihashi, and T. Fukuma, "High-speed atomic force microscopy for nano-visualization of dynamic biomolecular processes", Prog. Surf. Sci. 83: 337-437 (2008).

D. Yamamoto, T. Uchihashi, N. Kodera, and T. Ando, "Anisotropic diffusion of point defects in two-dimensional crystal of streptavidin observed by high-speed atomic force microscopy", Nanotechnol. 19: 384009 (9 pp) (2008).  (Supplementary Movie)  

A. Miyagi, Y. Tsunaka, T. Uchihashi, K. Mayanagi, S. Hirose, K. Morikawa, and T. Ando, "Visualization of intrinsically disordered regions of proteins by high-speed atomic force microscopy", Chem. Phys. Chem. 9(13):1859-1866 (2008).  (Supplementary Movies)

T. Fukuma, Y. Okazaki, N. Kodera, T. Uchihashi, and T. Ando, "High resonance frequency force microscope scanner using inertia balance support", Appl. Phys. Lett. 92:243119 (2008).

T. Ando, T. Uchihashi, N. Kodera, D. Yamamoto, M. Taniguchi, A. Miyagi, and H. Yamashita, "Invited Review: High-speed AFM and nano-visualization of biomolecular processes", Pflügers Archiv -Eur. J. Physiol. 456:211-225 (2008). 

T. Ando, T. Uchihashi, N. Kodera, D. Yamamoto, M. Taniguchi, A. Miyagi, and H. Yamashita, "Review: High-speed atomic force microscopy for observing dynamic biomolecular processes", J. Mol. Recognit. 20:448-458 (2007).  

H. Yamashita, T. Uchihashi, N. Kodera, A. Miyagi, D. Yamamoto, and T. Ando, "Tip-sample distance control using photo-thermal actuation of a small cantilever for high-speed atomic force microscopy", Rev. Sci. Instrum. 78:083702 (5 pages) (2007).

T. Uchihashi, H. Yamashita, and T. Ando, "Fast phase imaging in liquids using a rapid scan atomic force microscope.

Appl. Phys. Lett. 89:213112 (3 pages) (2006).

N. Kodera, M. Sakashita, and T. Ando, "Dynamic proportional-integral-differential controller for high-speed atomic force microscopy", Rev. Sci. Instrum. 77(8):083704 (7 pages) (2006).

M. Yokokawa, C. Wada, T. Ando, N. Sakai , A. Yagi, S.H. Yoshimura, and K. Takeyasu, "Fast-scanning atomic force microscopy reveals the ATP/ADP-dependent conformational changes of GroEL", EMBO J. 25:4567-4576 (2006). 

T. Ando, T. Uchihashi, N. Kodera, A. Miyagi, R. Nakakita, H. Yamashita, and M. Sakashita, "High-speed atomic force microscopy for studying the dynamic behavior of protein molecules at work", Jpn. J. Appl. Phys. 45(3B):1897-1903 (2006).

T. Uchihashi, N. Kodera, H. Itoh, H. Yamashita, and T. Ando, "Feed-forward control for high-speed AFM imaging of biomolecules", Jpn. J. Appl. Phys. 45(3B):1904-1908 (2006).

T. Ando, T. Uchihashi, N. Kodera, A. Miyagi, R. Nakakita, H. Yamashita, and K. Matada, "High-speed AFM for studying the dynamic behavior of protein molecules at work", Surf. Sci. Nanotechnol. 3:384-392 (2005).  (Supplementary Data)

N. Kodera, H. Yamashita, and T. Ando, "Active damping of the scanner for high-speed atomic force microscopy", Rev. Sci. Instrum. 76: 053708 (5pages) (2005). (Supplementary Data)

T. Ando, N. Kodera, Y. Naito, T. Kinoshita, K. Furuta, and Y.Y. Toyoshima, "A high-speed atomic force microscope for studying biological macromolecules in action", Chem. Phys. Chem. 4:1196-1202 (2003).

T. Ando, N. Kodera, D. Maruyama,  E. Takai, K. Saito, and A. Toda, "A High-speed atomic force microscope for studying biological macromolecules in action", Jpn. J. Appl. Phys. 41:4851-4856 (2002).

T. Ando, N. Kodera, E. Takai, D. Maruyama,  K. Saito, and A. Toda, "A High-speed atomic force microscope for studying biological macromolecules", Proc. Natl. Acad. Sci. USA  98:12468-12472 (2001).

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