Abstract
Atomic force microscopy (AFM) can probe single living cells and single native membrane proteins in natural fluid environments with label-free high spatial resolution. It has thus become an important tool for cellular and molecular biology that significantly complements traditional biochemical and biophysical techniques such as optical and electron microscopy and X-ray crystallography. Imaging surface topography is the primary application of AFM in the life sciences. Since the early 1990s, researchers have used AFM to investigate morphological features of living cells and native membrane proteins with impressive results. Steady improvements in AFM techniques for imaging soft biological samples have greatly expanded its applications. Based on the authors’ own research in AFM imaging of living cell morphologies, a review of sample preparation procedures for single-cell and single-molecule imaging experiments is presented, along with a summary of recent progress in AFM imaging of living cells and native membrane proteins. Finally, the challenges of AFM high-resolution imaging at the single-cell and single-molecule levels are discussed.
Article PDF
Similar content being viewed by others
Explore related subjects
Discover the latest articles and news from researchers in related subjects, suggested using machine learning.Avoid common mistakes on your manuscript.
References
Binnig G, Quate C F, Gerber C. Atomic force microscope. Phys Rev Lett, 1986, 56: 930–933
Katan A J, Dekker C. High-speed AFM reveals the dynamics of single biomolecules at the nanometer scale. Cell, 2011, 147: 979–982
Hinterdorfer P, Garcia-Parajo M F, Dufrene Y F. Single-molecule imaging of cell surfaces using near-field nanoscopy. Acc Chem Res, 2012, 45: 327–336
Muscariello L, Rosso F, Marino G, et al. A critical overview of ESEM applications in the biological field. J Cell Physiol, 2005, 205: 328–334
Kodera N, Yamamoto D, Ishikawa R, et al. Video imaging of walking myosin V by high-speed atomic force microscopy. Nature, 2010, 468: 72–76
Robertson J W F, Kasianowicz J J, Banerjee S. Analytical approaches for studying transporters, channels and porins. Chem Rev, 2012, 112: 6227–6249
Muller D J, Dufrene Y F. Atomic force microscopy as a multifunctional molecular toolbox in nanobiotechnology. Nat Nanotechnol, 2008, 3: 261–269
Kirat K E, Burton I, Dupres V, et al. Sample preparation procedures for biological atomic force microscopy. J Microsc, 2005, 218: 199–207
Matzke R, Jacobson K, Radmacher M. Direct, high-resolution measurement of furrow stiffening during division of adherent cells. Nat Cell Biol, 2001, 3: 607–610
Puntheeranurak T, Wildling L, Gruber H J, et al. Ligands on the string: Single-molecule AFM studies on the interaction of antibodies and substrates with the Na+-glucose co-transporter SGLT1 in living cells. J Cell Sci, 2006, 119: 2960–2967
Dufrene Y F. Atomic force microscopy and chemical force microscopy of microbial cells. Nat Protoc, 2008, 3: 1132–1138
Rosenbluth M J, Lam W A, Fletcher D A. Force microscopy of nonadherent cells: A comparison of leukemia cell deformability. Biophys J, 2006, 90: 2994–3003
Li M, Liu L, Xi N, et al. Imaging and measuring the rituximab-induced changes of mechanical properties in B-lymphoma cells using atomic force microscopy. Biochem Biophys Res Commun, 2011, 404: 689–694
Li M, Liu L, Xi N, et al. Drug-induced changes of topography and elasticity in living B lymphoma cells based on atomic force microscopy. Acta Phys Chim Sin, 2012, 28: 1502–1508
Mari S A, Pessoa J, Altieri S, et al. Gating of the MlotiK1 potassium channel involves large rearrangements of the cyclic nucleotide-binding domains. Proc Natl Acad Sci USA, 2011, 108: 20802–20807
Fotiadis D. Atomic force microscopy for the study of membrane proteins. Curr Opin Biotechnol, 2012, 23: 510–515
Muller D J, Engel A. Atomic force microscopy and spectroscopy of native membrane proteins. Nat Protoc, 2007, 2: 2191–2197
Muller D J. Adsorption of biological molecules to a solid support for scanning probe microscopy. J Struct Biol, 1997, 119: 172–188
Muller D J, Hand G M, Engel A, et al. Conformational changes in surface structures of isolated connexin 26 gap junctions. EMBO J, 2002, 14: 3598–3607
Radmacher M, Tillmann R W, Fritz M, et al. From molecules to cells: Imaging soft samples with the atomic force microscope. Science, 1992, 257: 1900–1905
Henderson E, Haydon P G, Sakaguchi D S. Actin filament dynamics in living glial cells imaged by atomic force microscopy. Science, 1992, 257: 1944–1946
Vie V, Giocondi M C, Lesniewska E, et al. Tapping-mode atomic force microscopy on intact cells: Optimal adjustment of tapping conditions by using the deflection signal. Ultramicroscopy, 2000, 82: 279–288
Chen P P, Dong H T, Chen L, et al. Application of atomic force microscopy to living samples from cells to fresh tissues. Chin Sci Bull, 2009, 54: 2410–2415
Alsteens D, Dupres V, Yunus S, et al. High-resolution imaging of chemical and biological sites on living cells using peak force tapping atomic force microscopy. Langmuir, 2012, 28: 16738–16744
Fantner G E, Barbero R J, Gray D S, et al. Kinetics of antimicrobial peptide activity measured on individual bacterial cells using high-speed atomic force microscopy. Nat Nanotechnol, 2010, 5: 280–285
Kirmse R, Otto H, Ludwig T. Interdependency of cell adhesion, force generation and extracellular proteolysis in matrix remodeling. J Cell Sci, 2011, 124: 1857–1866
El-Kirt-Chatel S, Dufrene Y F. Nanoscale imaging of the candidamacrophage interaction using correlated fluorescence-atomic force microscopy. ACS Nano, 2012, 6: 10792–10799
Baker M. Making membrane proteins for structures a trillion tiny tweaks. Nat Methods, 2010, 7: 429–433
Bill R M, Henderson P J F, Iwata S, et al. Overcoming barriers to membrane protein structure determination. Nat Biotechnol, 2011, 29: 335–340
Zhang X, Ren W, Decaen P, et al. Crystal structure of an orthologue of the NaChBac voltage-gated sodium channel. Nature, 2012, 486: 130–134
Ma D, Lu P, Yan C, et al. Structure and mechanism of a glutamate-GABA antiporter. Nature, 2012, 483: 632–636
Muller D J, Schabert F A, Buldt G, et al. Imaging purple membranes in aqueous solutions at sub-nanometer resolution by atomic force microscopy. Biophys J, 1995, 68: 1681–1686
Fotiadis D, Liang Y, Filipek S, et al. Rhodopsin dimers in native disc membranes. Nature, 2003, 421: 127–128
Shibata M, Yamashita H, Uchihashi T, et al. High-speed atomic force microscopy shows dynamic molecular processes in photoactivated bacteriorhodopsin. Nat Nanotechnol, 2010, 5: 208–212
Casuso I, Sens P, Rico F, et al. Experimental evidence for membrane-mediated protein-protein interaction. Biophys J, 2010, 99: L47–L49
Li M, Liu L Q, Xi N, et al. Atomic force microscopy imaging and mechanical properties measurement of red blood cells and aggressive cancer cells. Sci China Life Sci, 2012, 55: 968–973
Li M, Liu L Q, Xi N, et al. Detecting CD20-Rituximab interaction forces using AFM single-molecule force spectroscopy. Chin Sci Bull, 2011, 56: 3829–3835
Li M, Liu L Q, Xi N, et al. Imaging and mechanical property measurement of the lymphoma cells by atomic force microscopy (in Chinese). Chin Sci Bull (Chin Ver), 2010, 55: 2188–2196
Li M, Xiao X, Liu L, et al. Imaging and measuring the molecular force of lymphoma pathological cells using atomic force microscopy. Scanning, 2013, 35: 40–46
Li M, Liu L Q, Xi N, et al. Mapping CD20 molecules on the lymphoma cell surface using atomic force microscopy. Chin Sci Bull, 2013, 58: 1516–1519
Muller D J, Dufrene Y F. Force nanoscopy of living cells. Curr Biol, 2011, 21: R212–R216
Gross L, Mohn F, Moll N, et al. The chemical structure of a molecule resolved by atomic force microscopy. Science, 2009, 325: 1110–1114
Casuso I, Rico F, Sheuring S. High-speed atomic force microscopy: Structure and dynamics of single proteins. Curr Opin Chem Biol, 2011, 15: 704–709
Heinisch J J, Lipke P N, Beaussart A, et al. Atomic force microscopy-looking at mechanosensors on the cell surface. J Cell Sci, 2012, 125: 4189–4195
Dufrene Y F, Evans E, Engel A, et al. Five challenges to bringing single-molecule force spectroscopy into live cells. Nat Methods, 2011, 8: 123–127
Zhang P C, Keleshian A M, Sachs F. Voltage-induced membrane movement. Nature, 2001, 413: 428–432
Shekhawat G S, Dravid V P. Nanoscale imaging of buried structures via scanning near-field ultrasound holography. Science, 2005, 310: 89–92
Author information
Authors and Affiliations
Corresponding authors
Additional information
This article is published with open access at Springerlink.com
Rights and permissions
This article is published under an open access license. Please check the 'Copyright Information' section either on this page or in the PDF for details of this license and what re-use is permitted. If your intended use exceeds what is permitted by the license or if you are unable to locate the licence and re-use information, please contact the Rights and Permissions team.
About this article
Cite this article
Li, M., Liu, L., Xi, N. et al. Progress of AFM single-cell and single-molecule morphology imaging. Chin. Sci. Bull. 58, 3177–3182 (2013). https://doi.org/10.1007/s11434-013-5906-z
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11434-013-5906-z
