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Imaging modes of atomic force microscopy for application in molecular and cell biology

Abstract

Atomic force microscopy (AFM) is a powerful, multifunctional imaging platform that allows biological samples, from single molecules to living cells, to be visualized and manipulated. Soon after the instrument was invented, it was recognized that in order to maximize the opportunities of AFM imaging in biology, various technological developments would be required to address certain limitations of the method. This has led to the creation of a range of new imaging modes, which continue to push the capabilities of the technique today. Here, we review the basic principles, advantages and limitations of the most common AFM bioimaging modes, including the popular contact and dynamic modes, as well as recently developed modes such as multiparametric, molecular recognition, multifrequency and high-speed imaging. For each of these modes, we discuss recent experiments that highlight their unique capabilities.

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Figure 1: Timeline of key inventions, starting from the birth of AFM in 1986 to the latest AFM imaging modes in molecular and cell biology.
Figure 2: AFM-based imaging of native biological systems to molecular resolution.
Figure 3: Force–distance curve-based AFM.
Figure 4: Multifrequency AFM.
Figure 5: HS-AFM filming proteins in action.
Figure 6: AFM of cellular systems.

References

  1. 1

    Binnig, G., Quate, C. F. & Gerber, C. Atomic force microscope. Phys. Rev. Lett. 56, 930–933 (1986). This paper first described the principles of AFM.

    CAS  Article  Google Scholar 

  2. 2

    Gerber, C. & Lang, H. P. How the doors to the nanoworld were opened. Nat. Nanotech. 1, 3–5 (2006).

    CAS  Google Scholar 

  3. 3

    Binnig, G., Gerber, C., Stoll, E., Albrecht, T. R. & Quate, C. F. Atomic resolution with atomic force microscope. Europhys. Lett. 3, 1281–1286 (1987).

    CAS  Google Scholar 

  4. 4

    Drake, B. et al. Imaging crystals, polymers, and processes in water with the atomic force microscope. Science 243, 1586–1589 (1989).

    CAS  Google Scholar 

  5. 5

    Radmacher, M., Tillmann, R. W. & Gaub, H. E. Imaging viscoelasticity by force modulation with the atomic force microscope. Biophys. J. 64, 735–742 (1993).

    CAS  Google Scholar 

  6. 6

    Horber, J. K. & Miles, M. J. Scanning probe evolution in biology. Science 302, 1002–1005 (2003).

    CAS  Google Scholar 

  7. 7

    Binnig, G. & Rohrer, H. In touch with atoms. Rev. Mod. Phys. 71, S324 (1999).

    CAS  Google Scholar 

  8. 8

    Muller, D. J. & Dufrene, Y. F. Atomic force microscopy as a multifunctional molecular toolbox in nanobiotechnology. Nat. Nanotech. 3, 261–269 (2008).

    Google Scholar 

  9. 9

    Muller, D. J. & Dufrene, Y. F. Atomic force microscopy: a nanoscopic window on the cell surface. Trends Cell Biol. 21, 461–469 (2011).

    Google Scholar 

  10. 10

    Hansma, H. G. & Hoh, J. H. Biomolecular imaging with the atomic force microscope. Annu. Rev. Biophys. Biomol. Struct. 23, 115–139 (1994).

    CAS  Google Scholar 

  11. 11

    Hinterdorfer, P. & Dufrene, Y. F. Detection and localization of single molecular recognition events using atomic force microscopy. Nat. Methods 3, 347–355 (2006).

    CAS  Google Scholar 

  12. 12

    Ando, T., Uchihashi, T. & Kodera, N. High-speed AFM and applications to biomolecular systems. Ann. Rev. Biophys. 42, 393–414 (2013).

    CAS  Google Scholar 

  13. 13

    Dufrene, Y. F., Martinez-Martin, D., Medalsy, I., Alsteens, D. & Muller, D. J. Multiparametric imaging of biological systems by force-distance curve–based AFM. Nat. Methods 10, 847–854 (2013).

    CAS  Google Scholar 

  14. 14

    Garcia, R. & Proksch, R. Nanomechancial mapping of soft matter by bimodal force microscopy. Eur. Polym. J. 49, 1897–1906 (2013).

    CAS  Google Scholar 

  15. 15

    Radmacher, M., Tillmann, R. W., Fritz, M. & Gaub, H. E. From molecules to cells: imaging soft samples with the atomic force microscope. Science 257, 1900–1905 (1992).

    CAS  Google Scholar 

  16. 16

    Henderson, E., Haydon, P. G. & Sakaguchi, D. S. Actin filament dynamics in living glial cells imaged by atomic force microscopy. Science 257, 1944–1946 (1992).

    CAS  Google Scholar 

  17. 17

    Hoh, J. H. & Schoenenberger, C. A. Surface morphology and mechanical properties of MDCK monolayers by atomic force microscopy. J. Cell Sci. 107, 1105–1114 (1994).

    Google Scholar 

  18. 18

    Hoh, J. H., Lal, R., John, S. A., Revel, J.-P. & Arnsdorf, M. F. Atomic force microscopy and dissection of gap junctions. Science 253, 1405–1408 (1991).

    CAS  Google Scholar 

  19. 19

    Mou, J., Yang, J. & Shao, Z. Atomic force microscopy of cholera toxin B-oligomers bound to bilayers of biologically relevant lipids. J. Mol. Biol. 248, 507–512 (1995).

    CAS  Google Scholar 

  20. 20

    Schabert, F. A., Henn, C. & Engel, A. Native Escherichia coli OmpF porin surfaces probed by atomic force microscopy. Science 268, 92–94 (1995).

    CAS  Google Scholar 

  21. 21

    Hansma, H. G. et al. Reproducible imaging and dissection of plasmid DNA under liquid with the atomic force microscope. Science 256, 1180–1184 (1992).

    CAS  Google Scholar 

  22. 22

    Egger, M. et al. Wet lipid protein membranes imaged at submolecular resolution by atomic force microscopy. J. Struct. Biol. 103, 89–94 (1990).

    CAS  Google Scholar 

  23. 23

    Zasadzinski, J. A., Viswanathan, R., Madsen, L., Garnaes, J. & Schwartz, D. K. Langmuir-Blodgett films. Science 263, 1726–1733 (1994).

    CAS  Google Scholar 

  24. 24

    Yang, J., Mou, J. X. & Shao, Z. F. Structure and stability of pertussis toxin studied by in situ atomic force microscopy. FEBS Lett. 338, 89–92 (1994).

    CAS  Google Scholar 

  25. 25

    Müller, D. J., Schabert, F. A., Büldt, G. & Engel, A. Imaging purple membranes in aqueous solutions at sub-nanometer resolution by atomic force microscopy. Biophys. J. 68, 1681–1686 (1995).

    Google Scholar 

  26. 26

    Müller, D. J., Engel, A., Carrascosa, J. & Veléz, M. The bacteriophage phi29 head–tail connector imaged at high resolution with atomic force microscopy in buffer solution. EMBO J. 16, 2547–2553 (1997).

    Google Scholar 

  27. 27

    Czajkowsky, D. M., Sheng, S. & Shao, Z. Staphylococcal alpha-hemolysin can form hexamers in phospholipid bilayers. J. Mol. Biol. 276, 325–330 (1998).

    CAS  Google Scholar 

  28. 28

    Seelert, H. et al. Proton powered turbine of a plant motor. Nature 405, 418–419 (2000).

    CAS  Google Scholar 

  29. 29

    Scheuring, S., Reiss-Husson, F., Engel, A., Rigaud, J. L. & Ranck, J. L. High-resolution AFM topographs of Rubrivivax gelatinosus light- harvesting complex LH2. EMBO J. 20, 3029–3035 (2001).

    CAS  Google Scholar 

  30. 30

    Fotiadis, D. et al. Atomic-force microscopy: rhodopsin dimers in native disc membranes. Nature 421, 127–128 (2003).

    CAS  Google Scholar 

  31. 31

    Goldsbury, C., Kistler, J., Aebi, U., Arvinte, T. & Cooper, G. J. Watching amyloid fibrils grow by time-lapse atomic force microscopy. J. Mol. Biol. 285, 33–39 (1999).

    CAS  Google Scholar 

  32. 32

    Bezanilla, M. et al. Motion and enzymatic degradation of DNA in the atomic force microscope. Biophys. J. 67, 2454–2459 (1994).

    CAS  Google Scholar 

  33. 33

    Grandbois, M., Clausen-Schaumann, H. & Gaub, H. Atomic force microscope imaging of phospholipid bilayer degradation by phospholipase A2. Biophys. J. 74, 2398–2404 (1998).

    CAS  Google Scholar 

  34. 34

    Muller, D. J. & Engel, A. Voltage and pH-induced channel closure of porin OmpF visualized by atomic force microscopy. J. Mol. Biol. 285, 1347–1351 (1999).

    CAS  Google Scholar 

  35. 35

    Müller, D. J., Hand, G. M., Engel, A. & Sosinsky, G. Conformational changes in surface structures of isolated connexin 26 gap junctions. EMBO J. 21, 3598–3607 (2002). This paper reports using AFM to image animal communication channels at work with high-resolution.

    Google Scholar 

  36. 36

    Stoffler, D., Goldie, K. N., Feja, B & Aebi, U. Calcium-mediated structural changes of native nuclear pore complexes monitored by time-lapse atomic force microscopy. J. Mol. Biol. 287, 741–752 (1999).

    CAS  Google Scholar 

  37. 37

    Czajkowsky, D. M., Hotze, E. M., Shao, Z. & Tweten, R. K. Vertical collapse of a cytolysin prepore moves its transmembrane beta-hairpins to the membrane. EMBO J. 23, 3206–3215 (2004).

    CAS  Google Scholar 

  38. 38

    Scheuring, S. & Sturgis, J. N. Chromatic adaptation of photosynthetic membranes. Science 309, 484–487 (2005).

    CAS  Google Scholar 

  39. 39

    Engel, A. & Muller, D. J. Observing single biomolecules at work with the atomic force microscope. Nat. Struct. Biol. 7, 715–718 (2000).

    CAS  Google Scholar 

  40. 40

    Albrecht, T. R., Grutter, P., Horne, D. & Rugar, D. Frequency-modulation detection using high-Q cantilevers for enhanced force microscope sensitivity. J. Appl. Phys. 69, 668–673 (1991).

    Google Scholar 

  41. 41

    Putman, C. A. J., Vanderwerf, K. O., Degrooth, B. G., Vanhulst, N. F. & Greve, J. Tapping mode atomic-force microscopy in liquid. Appl. Phys. Lett. 64, 2454–2456 (1994).

    CAS  Google Scholar 

  42. 42

    Garcia, R. & Herruzo, E. T. The emergence of multifrequency force microscopy. Nat. Nanotech. 7, 217–226 (2012). A review describing recent progress in multifrequency force microscopy, and discussing its potential for studying proteins and cells.

    CAS  Google Scholar 

  43. 43

    Hansma, P. K. et al. Tapping mode atomic-force microscopy in liquids. Appl. Phys. Lett. 64, 1738–1740 (1994).

    CAS  Google Scholar 

  44. 44

    Wegmann, S. et al. Human Tau isoforms assemble into ribbon-like fibrils that display polymorphic structure and stability. J. Biol. Chem. 285, 27302–27313 (2010).

    CAS  Google Scholar 

  45. 45

    Ido, S. et al. Beyond the helix pitch: direct visualization of native DNA in aqueous solution. ACS Nano 7, 1817–1822 (2013).

    CAS  Google Scholar 

  46. 46

    Ido, S. et al. Immunoactive two-dimensional self-assembly of monoclonal antibodies in aqueous solution revealed by atomic force microscopy. Nat. Mater. 13, 264–270 (2014).

    CAS  Google Scholar 

  47. 47

    Möller, C., Allen, M., Elings, V., Engel, A. & Müller, D. J. Tapping mode atomic force microscopy produces faithful high-resolution images of protein surfaces. Biophys. J. 77, 1050–1058 (1999).

    Google Scholar 

  48. 48

    Stark, M., Moller, C., Muller, D. J. & Guckenberger, R. From images to interactions: high-resolution phase imaging in tapping-mode atomic force microscopy. Biophys. J. 80, 3009–3018 (2001).

    CAS  Google Scholar 

  49. 49

    Kasas, S. & Ikai, A. A method for anchoring round shaped cells for atomic force microscope imaging. Biophys. J. 68, 1678–1680 (1995).

    CAS  Google Scholar 

  50. 50

    Andre, G. et al. Imaging the nanoscale organization of peptidoglycan in living Lactococcus lactis cells. Nat. Commun. 1, 27 (2010).

    Google Scholar 

  51. 51

    Hansma, P. K., Drake, B., Marti, O., Gould, S. A. & Prater, C. B. The scanning ion-conductance microscope. Science 243, 641–643 (1989).

    CAS  Google Scholar 

  52. 52

    Novak, P. et al. Nanoscale live-cell imaging using hopping probe ion conductance microscopy. Nat. Methods 6, 279–281 (2009).

    CAS  Google Scholar 

  53. 53

    Drake, B., Randall, C., Bridges, D. & Hansma, P. K. A new ion sensing deep atomic force microscope. Rev. Sci. Instrum. 85, 083706 (2014).

    Google Scholar 

  54. 54

    Roos, W. H., Bruinsma, R. & Wuite, G. J. L. Physical virology. Nat. Phys. 6, 733–743 (2010).

    CAS  Google Scholar 

  55. 55

    Oesterhelt, F. et al. Unfolding pathways of individual bacteriorhodopsins. Science 288, 143–146 (2000).

    CAS  Google Scholar 

  56. 56

    Kufer, S. K., Puchner, E. M., Gumpp, H., Liedl, T. & Gaub, H. E. Single-molecule cut-and-paste surface assembly. Science 319, 594–596 (2008).

    CAS  Google Scholar 

  57. 57

    Braunschweig, A. B., Huo, F. & Mirkin, C. A. Molecular printing. Nat. Chem. 1, 353–358 (2009).

    CAS  Google Scholar 

  58. 58

    Cattin, C. J. et al. Mechanical control of mitotic progression in single animal cells. Proc. Natl Acad. Sci. USA 112, 11258–11263 (2015).

    CAS  Google Scholar 

  59. 59

    Butt, H. J., Cappella, B. & Kappl, M. Force measurements with the atomic force microscope: technique, interpretation and applications. Surf. Sci. Rep. 59, 1–152 (2005).

    CAS  Google Scholar 

  60. 60

    Dong, M., Husale, S. & Sahin, O. Determination of protein structural flexibility by microsecond force spectroscopy. Nat. Nanotech. 4, 514–517 (2009).

    CAS  Google Scholar 

  61. 61

    Martinez-Martin, D., Herruzo, E. T., Dietz, C., Gomez-Herrero, J. & Garcia, R. Noninvasive protein structural flexibility mapping by bimodal dynamic force microscopy. Phys. Rev. Lett. 106, 198101 (2011).

    CAS  Google Scholar 

  62. 62

    Herruzo, E. T., Perrino, A. P. & Garcia, R. Fast nanomechanical spectroscopy of soft matter. Nat. Commun. 5, 3126 (2014).

    Google Scholar 

  63. 63

    Preiner, J. et al. High-speed AFM images of thermal motion provide stiffness map of interfacial membrane protein moieties. Nano Lett. 15, 759–763 (2015).

    CAS  Google Scholar 

  64. 64

    Radmacher, M., Cleveland, J. P., Fritz, M., Hansma, H. G. & Hansma, P. K. Mapping interaction forces with the atomic force microscope. Biophys. J. 66, 2159–2165 (1994).

    CAS  Google Scholar 

  65. 65

    Heinz, W. F. & Hoh, J. H. Spatially resolved force spectroscopy of biological surfaces using the atomic force microscope. Trends Biotechnol. 17, 143–150 (1999).

    CAS  Google Scholar 

  66. 66

    Rotsch, C. & Radmacher, M. Drug-induced changes of cytoskeletal structure and mechanics in fibroblasts: an atomic force microscopy study. Biophys. J. 78, 520–535 (2000).

    CAS  Google Scholar 

  67. 67

    Matzke, R., Jacobson, K. & Radmacher, M. Direct, high-resolution measurement of furrow stiffening during division of adherent cells. Nat. Cell Biol. 3, 607–610 (2001).

    CAS  Google Scholar 

  68. 68

    Plodinec, M. et al. The nanomechanical signature of breast cancer. Nat. Nanotech. 7, 757–765 (2012).

    CAS  Google Scholar 

  69. 69

    Rebelo, L. M., de Sousa, J. S., Mendes Filho, J. & Radmacher, M. Comparison of the viscoelastic properties of cells from different kidney cancer phenotypes measured with atomic force microscopy. Nanotechnology 24, 055102 (2013).

    CAS  Google Scholar 

  70. 70

    Touhami, A., Nysten, B. & Dufrêne, Y. F. Nanoscale mapping of the elasticity of microbial cells by atomic force microscopy. Langmuir 19, 4539–4543 (2003).

    CAS  Google Scholar 

  71. 71

    Viani, M. B. et al. Fast imaging and fast force spectroscopy of single biopolymers with a new atomic force microscope designed for small cantilevers. Rev. Sci. Instrum. 70, 4300–4303 (1999). This paper reports the invention of small cantilevers for fast AFM imaging and force spectroscopy.

    CAS  Google Scholar 

  72. 72

    Viani, M. B. et al. Small cantilevers for force spectroscopy of single molecules. J. Appl. Phys. 86, 2258–2262 (1999).

    CAS  Google Scholar 

  73. 73

    Ando, T. et al. A high-speed atomic force microscope for studying biological macromolecules. Proc. Natl Acad. Sci. USA 98, 12468–12472 (2001).

    CAS  Google Scholar 

  74. 74

    Alcaraz, J. et al. Correction of microrheological measurements of soft samples with atomic force microscopy for the hydrodynamic drag on the cantilever. Langmuir 18, 716–721 (2002).

    CAS  Google Scholar 

  75. 75

    Sahin, O., Magonov, S., Su, C., Quate, C. F. & Solgaard, O. An atomic force microscope tip designed to measure time-varying nanomechanical forces. Nat. Nanotech. 2, 507–514 (2007).

    Google Scholar 

  76. 76

    Medalsy, I., Hensen, U. & Muller, D. J. Imaging and quantifying chemical and physical properties of native proteins at molecular resolution by force-volume AFM. Angew. Chem. Int. Ed. 50, 12103–12108 (2011).

    CAS  Google Scholar 

  77. 77

    Sullan, R. M., Li, J. K. & Zou, S. Direct correlation of structures and nanomechanical properties of multicomponent lipid bilayers. Langmuir 25, 7471–7477 (2009).

    CAS  Google Scholar 

  78. 78

    Heu, C., Berquand, A., Elie-Caille, C. & Nicod, L. Glyphosate-induced stiffening of HaCaT keratinocytes, a Peak Force Tapping study on living cells. J. Struct. Biol. 178, 1–7 (2012).

    CAS  Google Scholar 

  79. 79

    Formosa-Dague, C., Speziale, P., Foster, T. J., Geoghegan, J. A. & Dufrene, Y. F. Zinc-dependent mechanical properties of Staphylococcus aureus biofilm-forming surface protein SasG. Proc. Natl Acad. Sci. USA 113, 410–415 (2016).

    CAS  Google Scholar 

  80. 80

    Alsteens, D., Trabelsi, H., Soumillion, P. & Dufrene, Y. F. Multiparametric atomic force microscopy imaging of single bacteriophages extruding from living bacteria. Nat. Commun. 4, 2926 (2013).

    Google Scholar 

  81. 81

    Carrasco, C. et al. Built-in mechanical stress in viral shells. Biophys. J. 100, 1100–1108 (2011).

    CAS  Google Scholar 

  82. 82

    Zink, M. & Grubmuller, H. Mechanical properties of the icosahedral shell of southern bean mosaic virus: a molecular dynamics study. Biophys. J. 96, 1350–1363 (2009).

    CAS  Google Scholar 

  83. 83

    Carrasco, C. et al. DNA-mediated anisotropic mechanical reinforcement of a virus. Proc. Natl Acad. Sci. USA 103, 13706–13711 (2006).

    CAS  Google Scholar 

  84. 84

    Alsteens, D., Garcia, M. C., Lipke, P. N. & Dufrene, Y. F. Force-induced formation and propagation of adhesion nanodomains in living fungal cells. Proc. Natl Acad. Sci. USA 107, 20744–20749 (2010). This paper reports using recognition imaging to demonstrate that microbial cell adhesion proteins form nanoclusters under mechanical force.

    CAS  Google Scholar 

  85. 85

    Pfreundschuh, M., Hensen, U. & Muller, D. J. Quantitative imaging of the electrostatic field and potential generated by a transmembrane protein pore at subnanometer resolution. Nano Lett. 13, 5585–5593 (2013).

    CAS  Google Scholar 

  86. 86

    Evans, E. A. & Calderwood, D. A. Forces and bond dynamics in cell adhesion. Science 316, 1148–1153 (2007).

    CAS  Google Scholar 

  87. 87

    Medalsy, I. D. & Muller, D. J. Nanomechanical properties of proteins and membranes depend on loading rate and electrostatic interactions. ACS Nano 7, 2642–2650 (2013).

    CAS  Google Scholar 

  88. 88

    Frisbie, C. D., Rozsnyai, L. F., Noy, A., Wrighton, M. S. & Lieber, C. M. Functional group imaging by chemical force microscopy. Science 265, 2071–2074 (1994).

    CAS  Google Scholar 

  89. 89

    Hinterdorfer, P., Baumgartner, W., Gruber, H. J., Schilcher, K. & Schindler, H. Detection and localization of individual antibody-antigen recognition events by atomic force microscopy. Proc. Natl Acad. Sci. USA 93, 3477–3481 (1996).

    CAS  Google Scholar 

  90. 90

    Ludwig, M., Dettmann, W. & Gaub, H. E. Atomic force microscope imaging contrast based on molecular recognition. Biophys. J. 72, 445–448 (1997).

    CAS  Google Scholar 

  91. 91

    Grandbois, M., Dettmann, W., Benoit, M. & Gaub, H. E. Affinity imaging of red blood cells using an atomic force microscope. J. Histochem. Cytochem. 48, 719–724 (2000).

    CAS  Google Scholar 

  92. 92

    Florin, E. L., Moy, V. T. & Gaub, H. E. Adhesion forces between individual ligand-receptor pairs. Science 264, 415–417 (1994).

    CAS  Google Scholar 

  93. 93

    Lee, G. U., Chrisey, L. A. & Colton, R. J. Direct measurement of the forces between complementary strands of DNA. Science 266, 771–773 (1994).

    CAS  Google Scholar 

  94. 94

    Kienberger, F. et al. Recognition force spectroscopy studies of the NTA-His6 bond. Single Mol. 1, 59–65 (2000).

    CAS  Google Scholar 

  95. 95

    Thie, M. et al. Interactions between trophoblast and uterine epithelium: monitoring of adhesive forces. Hum. Reprod. 13, 3211–3219 (1998).

    CAS  Google Scholar 

  96. 96

    Kim, H., Arakawa, H., Osada, T. & Ikai, A. Quantification of cell adhesion force with AFM: distribution of vitronectin receptors on a living MC3T3-E1 cell. Ultramicroscopy 97, 359–363 (2003).

    CAS  Google Scholar 

  97. 97

    Kim, H. et al. Quantification of the number of EP3 receptors on a living CHO cell surface by the AFM. Ultramicroscopy 106, 652–662 (2006).

    CAS  Google Scholar 

  98. 98

    Roduit, C. et al. Elastic membrane heterogeneity of living cells revealed by stiff nanoscale membrane domains. Biophys. J. 94, 1521–1532 (2008).

    CAS  Google Scholar 

  99. 99

    Alsteens, D. et al. Imaging G protein–coupled receptors while quantifying their ligand-binding free-energy landscape. Nat. Methods 12, 845–851 (2015). This paper showed that attaching a ligand to the AFM stylus allows it to image and map its binding to human G protein–coupled receptors and to reconstruct the ligand-binding free-energy landscape.

    CAS  Google Scholar 

  100. 100

    Andre, G. et al. Fluorescence and atomic force microscopy imaging of wall teichoic acids in Lactobacillus plantarum. ACS Chem. Biol. 6, 366–376 (2011).

    CAS  Google Scholar 

  101. 101

    Dupres, V. et al. Nanoscale mapping and functional analysis of individual adhesins on living bacteria. Nat. Methods 2, 515–520 (2005). This paper reports that AFM tips labelled with bioligands can map the distribution of single adhesion proteins on bacterial pathogens and reveal their assembly into nanodomains.

    CAS  Google Scholar 

  102. 102

    Pfreundschuh, M. et al. Identifying and quantifying two ligand-binding sites while imaging native human membrane receptors by AFM. Nat. Commun. 6, 8857 (2015).

    CAS  Google Scholar 

  103. 103

    Raab, A. et al. Antibody recognition imaging by force microscopy. Nat. Biotechnol. 17, 901–905 (1999).

    CAS  Google Scholar 

  104. 104

    Stroh, C. et al. Single-molecule recognition imaging microscopy. Proc. Natl Acad. Sci. USA 101, 12503–12507 (2004).

    CAS  Google Scholar 

  105. 105

    Chtcheglova, L. A., Waschke, J., Wildling, L., Drenckhahn, D. & Hinterdorfer, P. Nano-scale dynamic recognition imaging on vascular endothelial cells. Biophys. J. 93, L11–L13 (2007).

    CAS  Google Scholar 

  106. 106

    Zhang, S., Aslan, H., Besenbacher, F. & Dong. M. Quantitative biomolecular imaging by dynamic nanomechanical mapping. Chem. Soc. Rev. 43, 7412–7429 (2014).

    CAS  Google Scholar 

  107. 107

    Dietz, C., Herruzo, E. T., Lozano, J. R. & Garcia, R. Nanomechanical coupling enables detection and imaging of 5 nm superparamagnetic particles in liquid. Nanotechnology 22, 125708 (2011).

    Google Scholar 

  108. 108

    Herruzo, E. T., Asakawa, H., Fukuma, T. & Garcia, R. Three-dimensional quantitative force maps in liquid with 10 piconewton, angstrom and sub-minute resolutions. Nanoscale 5, 2678–2685 (2013).

    CAS  Google Scholar 

  109. 109

    Fukuma, T., Higgins, M. J. & Jarvis, S. P. Direct imaging of individual intrinsic hydration layers on lipid bilayers at Angstrom resolution. Biophys. J. 92, 3603–3609 (2007).

    CAS  Google Scholar 

  110. 110

    Cartagena, A., Hernando-Perez, M., Carrascosa, J. L., de Pablo, P. J. & Raman, A. Mapping in vitro local material properties of intact and disrupted virions at high resolution using multi-harmonic atomic force microscopy. Nanoscale 5, 4729–4736 (2013).

    CAS  Google Scholar 

  111. 111

    Cartagena-Rivera, A. X., Wang, W. H., Geahlen, R. L. & Raman, A. Fast, multi-frequency, and quantitative nanomechanical mapping of live cells using the atomic force microscope. Sci. Rep. 5, 11692 (2015).

    CAS  Google Scholar 

  112. 112

    Kim, D. & Sahin, O. Imaging and three-dimensional reconstruction of chemical groups inside a protein complex using atomic force microscopy. Nat. Nanotech. 10, 264–269 (2015).

    CAS  Google Scholar 

  113. 113

    Shekhawat, G. S. & Dravid, V. P. Nanoscale imaging of buried structures via scanning near-field ultrasound holography. Science 310, 89–92 (2005).

    CAS  Google Scholar 

  114. 114

    Tetard, L. et al. Imaging nanoparticles in cells by nanomechanical holography. Nat. Nanotech. 3, 501–505 (2008).

    CAS  Google Scholar 

  115. 115

    Verbiest, G. J. & Rost, M. J. Beating beats mixing in heterodyne detection schemes. Nat. Commun. 6, 6444 (2015).

    CAS  Google Scholar 

  116. 116

    Kindt, J. H., Fantner, G. E., Cutroni, J. A. & Hansma, P. K. Rigid design of fast scanning probe microscopes using finite element analysis. Ultramicroscopy 100, 259–265 (2004).

    CAS  Google Scholar 

  117. 117

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

    CAS  Google Scholar 

  118. 118

    Kodera, M., Yamashita, H. & Ando, T. Active damping of the scanner for high-speed atomic force microscopy. Rev. Sci. Instrum. 76, 053708 (2005).

    Google Scholar 

  119. 119

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

    Google Scholar 

  120. 120

    Viani, M. B. et al. Probing protein–protein interactions in real time. Nat. Struct. Biol. 7, 644–647 (2000).

    CAS  Google Scholar 

  121. 121

    Ando, T. et al. A high-speed atomic force microscope for studying biological macromolecules in action. ChemPhysChem 4, 1196–1202 (2003).

    CAS  Google Scholar 

  122. 122

    Shibata, M., Yamashita, H., Uchihashi, T., Kandori, H. & Ando, T. High-speed atomic force microscopy shows dynamic molecular processes in photoactivated bacteriorhodopsin. Nat. Nanotech. 5, 208–212 (2010).

    CAS  Google Scholar 

  123. 123

    Kodera, N., Yamamoto, D., Ishikawa, R. & Ando, T. Video imaging of walking myosin V by high-speed atomic force microscopy. Nature 468, 72–76 (2010). This paper showed that high-speed AFM can be used to watch proteins functioning in real-time.

    CAS  Google Scholar 

  124. 124

    Uchihashi, T., Iino, R., Ando, T. & Noji, H. High-speed atomic force microscopy reveals rotary catalysis of rotorless F(1)-ATPase. Science 333, 755–758 (2011).

    CAS  Google Scholar 

  125. 125

    Chiaruttini, N. et al. Relaxation of loaded ESCRT-III spiral springs drives membrane deformation. Cell 163, 866–879 (2015).

    CAS  Google Scholar 

  126. 126

    Sakiyama, Y., Mazur, A., Kapinos, L. E. & Lim, R. Y. Spatiotemporal dynamics of the nuclear pore complex transport barrier resolved by high-speed atomic force microscopy. Nat. Nanotech. 11, 719–723 (2016).

    CAS  Google Scholar 

  127. 127

    Fantner, G. E., Barbero, R. J., Gray, D. S. & Belcher, A. M. Kinetics of antimicrobial peptide activity measured on individual bacterial cells using high-speed atomic force microscopy. Nat. Nanotech. 5, 280–285 (2010).

    CAS  Google Scholar 

  128. 128

    Yamashita, H. et al. Single-molecule imaging on living bacterial cell surface by high-speed AFM. J. Mol. Biol. 422, 300–309 (2012).

    CAS  Google Scholar 

  129. 129

    Shibata, M., Uchihashi, T., Ando, T. & Yasuda, R. Long-tip high-speed atomic force microscopy for nanometer-scale imaging in live cells. Sci. Rep. 5, 8724 (2015).

    CAS  Google Scholar 

  130. 130

    Uchihashi, T., Watanabe, H., Fukuda, S., Shibata, M. & Ando, T. Functional extension of high-speed AFM for wider biological applications. Ultramicroscopy 160, 182–196 (2016).

    CAS  Google Scholar 

  131. 131

    El-Kirat-Chatel, S. & Dufrene, Y. F. Nanoscale imaging of the Candida — macrophage interaction using correlated fluorescence-atomic force microscopy. ACS Nano 6, 10792–10799 (2012).

    CAS  Google Scholar 

  132. 132

    Sharma, A., Anderson, K. & Muller, D. J. Actin microridges characterized by laser scanning confocal and atomic force microscopy. FEBS Lett. 579, 2001–2009 (2005).

    CAS  Google Scholar 

  133. 133

    Schillers, H., Medalsy, I., Hu, S., Slade, A. L. & Shaw, J. E. PeakForce Tapping resolves individual microvilli on living cells. J. Mol. Recognit. 29, 95–101 (2016).

    CAS  Google Scholar 

  134. 134

    Benoit, M., Gabriel, D., Gerisch, G. & Gaub, H. E. Discrete interactions in cell adhesion measured by single-molecule force spectroscopy. Nat. Cell Biol. 2, 313–317 (2000).

    CAS  Google Scholar 

  135. 135

    Krieg, M. et al. Tensile forces govern germ-layer organization in zebrafish. Nat. Cell Biol. 10, 429–436 (2008).

    CAS  Google Scholar 

  136. 136

    Cuerrier, C. M., Gagner, A., Lebel, R., Gobeil, F. Jr & Grandbois, M. Effect of thrombin and bradykinin on endothelial cell mechanical properties monitored through membrane deformation. J. Mol. Recognit. 22, 389–396 (2009).

    CAS  Google Scholar 

  137. 137

    Pelling, A. E., Veraitch, F. S., Chu, C. P., Mason, C. & Horton, M. A. Mechanical dynamics of single cells during early apoptosis. Cell Motil. Cytoskel. 66, 409–422 (2009).

    CAS  Google Scholar 

  138. 138

    Ramanathan, S. P. et al. Cdk1-dependent mitotic enrichment of cortical myosin II promotes cell rounding against confinement. Nat. Cell Biol. 17, 148–159 (2015).

    CAS  Google Scholar 

  139. 139

    Stewart, M. P. et al. Hydrostatic pressure and the actomyosin cortex drive mitotic cell rounding. Nature 469, 226–230 (2011).

    CAS  Google Scholar 

  140. 140

    Duman, M. et al. Improved localization of cellular membrane receptors using combined fluorescence microscopy and simultaneous topography and recognition imaging. Nanotechnology 21, 115504 (2010).

    CAS  Google Scholar 

  141. 141

    Lipke, P. N. et al. Strengthening relationships: amyloids create adhesion nanodomains in yeasts. Trends Microbiol. 20, 59–65 (2012).

    CAS  Google Scholar 

  142. 142

    Alsteens, D. et al. Nanomechanical mapping of first binding steps of a virus to animal cells. Nat. Nanotech. 12, 177–183 (2017). This paper showed that attaching a rabies virus to the AFM stylus allows living animal cells to be imaged with confocal microscopy and AFM, to simultaneously localize virus-binding, and to quantify the virus-binding process and free-energy landscape.

    CAS  Google Scholar 

  143. 143

    Churnside, A. B. & Perkins, T. T. Ultrastable atomic force microscopy: improved force and positional stability. FEBS Lett. 588, 3621–3630 (2014).

    CAS  Google Scholar 

  144. 144

    King, G. M., Carter, A. R., Churnside, A. B., Eberle, L. S. & Perkins, T. T. Ultrastable atomic force microscopy: atomic-scale stability and registration in ambient conditions. Nano Lett. 9, 1451–1456 (2009).

    CAS  Google Scholar 

  145. 145

    Franz, C. M. & Muller, D. J. Analysing focal adhesion structure by AFM. J. Cell Sci. 118, 5315–5323 (2005).

    CAS  Google Scholar 

  146. 146

    Lucas, R. W., Kuznetsov, Y. G., Larson, S. B. & McPherson, A. Crystallization of Brome mosaic virus and T = 1 Brome mosaic virus particles following a structural transition. Virology 286, 290–303 (2001).

    CAS  Google Scholar 

  147. 147

    Friedrichs, J., Taubenberger, A., Franz, C. M. & Muller, D. J. Cellular remodelling of individual collagen fibrils visualized by time-lapse AFM. J. Mol. Biol. 372, 594–607 (2007).

    CAS  Google Scholar 

  148. 148

    Mari, S. A. et al. Gating of the MlotiK1 potassium channel involves large rearrangements of the cyclic nucleotide-binding domains. Proc. Natl Acad. Sci. USA 108, 20802–20807 (2011).

    Google Scholar 

  149. 149

    Bestembayeva, A. et al. Nanoscale stiffness topography reveals structure and mechanics of the transport barrier in intact nuclear pore complexes. Nat. Nanotech. 10, 60–64 (2015).

    CAS  Google Scholar 

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Acknowledgements

Y.F.D. was supported by the Université catholique de Louvain, the European Research Council (ERC) under the European Union's Horizon 2020 research and innovation programme (grant agreement no. 693630), the WELBIO (grant no. WELBIO-CR-2015A-05), the National Fund for Scientific Research (FNRS), the Federal Office for Scientific, Technical and Cultural Affairs (Interuniversity Poles of Attraction Programme) and the Research Department of the Communauté française de Belgique (Concerted Research Action). D.A. and D.M.M. were supported by the European Molecular Biology Organization (EMBO; ALTF 265-2013 and ALTF 506-2012). D.J.M. was supported by the Swiss National Science Foundation (SNF; grants 205320_160199 and 310030B_160225), the NCCR Molecular Systems Engineering and the Swiss Commission for Technology and Innovation (CTI; grant 17970.1). C.G. was supported by the Swiss Nano Institute (SNI) of the University of Basel. R.G. acknowledges financial support from the European Research Council AdG no. 340177 and the Ministerio de Economia y Competitividad MAT2016-76507-R. T.A. was supported by the Japan Society for the Promotion of Science (JSPS; grants 24227005 and 26119003) and by the Japan Science and Technology Agency (JST; CREST program on Structural Life Science and Advanced Core Technology for Innovative Life Science Research).

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Correspondence to Yves F. Dufrêne or Daniel J. Müller.

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Dufrêne, Y., Ando, T., Garcia, R. et al. Imaging modes of atomic force microscopy for application in molecular and cell biology. Nature Nanotech 12, 295–307 (2017). https://doi.org/10.1038/nnano.2017.45

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