Abstract
The investigative power of a single technique can be significantly enhanced by the simultaneous addition of a second, independent technique. This is especially true for the combination of atomic force microscopy (AFM) and patch-clamp recording for the study of biological systems because each technique has the ability to observe dynamic molecular events under physiological conditions. Merging these approaches will have direct application in examining structures in which mechanical and electrical parameters are paramount; for example, the study of mechanosensitive ion channels (MSCs), flexoelectricity (mechanical/electrical coupling), and outer-hair-cell electromotility. In addition, subtle advantages of this arrangement can be beneficial in studying related problems of membrane mechanics and voltage- and ligand-sensitive ion-channel physiology. Correlating mechanical and electrical interactions on a molecular level has only been attempted in a few instances. This chapter introduces our attempt to create a setup of standard components that allows concurrent use of both methods without limiting the practical use of either technique individually. We discuss experimental results exploiting this setup and consider the possibilities for further applications. We will begin with a review of some history of use of AFM and patch-clamp (summarized in Table 1)
Keywords
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Hille, B. (1992) Ionic Channels of Excitable Membranes. Sinauer Associates Inc., Sunderland, MA.
Horn, R. (1991) Diffusion of nystatin in plasma membrane is inhibited by a glass-membrane seal [published erratum appears in Biophys. J. 60(4), 985]. Biophys. J. 60, 329–333.
Fernandez, J. M., Neher, E., and Gomperts, B. D. (1984) Capacitance measurements reveal stepwise fusion events in degranulating mast cells. Nature 312,5993), 453–455.
Joshi, C. and Fernandez, J. M. (1989) Capacitance measurements: an analysis of the phase detector technique used to study exocytosis and endocytosis. Biophys. J. 56, 1153–1162.
Dilger, J. P., Brett, R. S., Poppers, D. M., and Liu, Y. (1991) The temperature dependence of some kinetic and conductance properties of acetylcholine receptor channels. Biochim. Biophys. Acta 1063, 253–258.
Sachs, F. (1999) Practical limits on the maximal speed of solution exchange for patch-clamp experiments. Biophys J. 77, 682–690.
Besch, S. D. and Sachs, F. (2000) A Compact, High Speed, Air Pressure Servo. In press.
Mcbride, D. W. and Hamill, O. P. (1995) A fast pressure-clamp technique for studying mechanogated channels, in Single-Channel Recording (Sakmann, B. and Neher, E., eds.), Plenum, New York, pp. 329–340.
Martinac, B. (1993) Mechanosensitive ion channels: biophysics and physiology. Thermodynamics of Cell Surface Receptors (Jackson, M., ed.), CRC Press, Boca Raton, FL, pp. 327–351.
Sachs, F. (1992) Stretch sensitive ion channels: an update, in Sensory Transduction (Corey, D. P. and Roper, S. D., eds.), Rockefeller University Press, Society General Physiology, NY, pp. 241–260.
Sackin, H. (1995) Stretch activated ion channels. Kidney Int. 48, 1134–1147.
Sachs, F. and Morris, C. (1998) Mechanosensitive ion channels in non specialized cells, in Reviews of Physiology and Biochemistry and Pharmacology (Blaustein, M. P., et al., eds.), Springer, Berlin, pp. 1–78.
Sokabe, M. and Sachs, F. (1992) Towards a molecular mechanism of activation in mechanosensitive ion channels, in Advances in Comparative and Environmental Physiology, vol. 10 (Ito, F., ed.), Springer-Verlag, Berlin, pp. 55–77.
Sukharev, S., Sigurdson, W., Kung, C., and Sachs, F. (1999) Energetic and spatial parameters for gating of the bacterial large conductance mechanosensitive channel, MscL. J. Gen. Physiol. 113, 525–539.
Hamill, O. P., Marty, A., Neher, E., Sakmann, B., and Sigworth, F. J. (1981) Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflugers Arch. Eur. J. Physiol. 391, 85–100.
Guharay, F. and Sachs, F. (1984) Stretch-activated single ion channel currents in tissue-cultured embryonic chick skeletal muscle. J. Physiol. (Lond.) 352, 685–701.
Yang, X. C. and Sachs, F. (1989) Block of stretch-activated ion channels in Xenopus oocytes by gadolinium and calcium ions. Science 243, 1068–1071.
Sokabe, M., Sachs, F., and Jing, Z. (1991) Quantitative video microscopy of patch clamped membranes: stress, strain, capacitance and stretch channel activation. Biophys. J. 59, 722–728.
Small, D. L. and Morris, C. E. (1994) Delayed activation of single mechanosensitive channels in Lymnaea neurons. Am. J. Physiol. 267, C598–C606.
Hamill, O. P. and McBride, D. W., Jr. (1992) Rapid adaptation of single mechanosensitive channels in Xenopus oocytes. Proc. Natl. Acad. Sci. USA 89, 7462–7466.
Suchyna, T. M., Johnson, J. H., Clemo, H. F., Huang, Z. H., Gage, D. A., Baumgarten, C. M., and Sachs, F. (2000) Identification of a peptide toxin from Grammostola spatulata spider venom that blocks stretch activated channels. J. Gen. Physiol. 115, 583–598.
Fahlke, C. and Rudel, R. (1992) Giga-seal formation alters properties of sodium channels of human myoballs. Pflugers Arch. 420, 248–254.
Hochmuth, R. M. (2000) Micropipette aspiration of living cells. [Review] [34 refs]. J. Biomechanics 33, 15–22.
Horber, J. K., Mosbacher, J., Haberle, W., Ruppersberg, J. P., and Sakmann, B. (1995) A look at membrane patches with a scanning force microscope. Biophys. J. 68, 1687–1693.
Drake, B. Prater, C. B. Weisenhorn, A. L., Gould, S. A., Albrecht, T. R., Quate, C. F., et al. (1989) Imaging crystals, polymers, and processes in water with the atomic force microscope. Science 243, 1586–1589.
Radmacher, M., Hillner, P. E., and Hansma, P. K. (1994) Scanning nearfield optical microscope using microfabricated probes. Rev. Sci. Instrum. 65, 2737–2738.
Hansma, H. and Hoh, J. (1994) Biomolecular imaging with the atomic force microscope. Ann. Rev. Biophys. Biomol. Struct. 23, 115–139.
Lal, R., Kim, H., Garavito, R. M., and Arnsdorf, M. F. (1993) Imaging of reconstituted biological channels at molecular resolution by atomic force microscopy. Am. J. Physiol. 265, C851–C856.
Jena, B. P., Schneider, S. W., Geibel, J. P., Webster, P., Oberleithner, H., and Sritharan, K. C. (1997) Gi regulation of secretory vesicle swelling examined by atomic force microscopy. Proc. Natl. Acad. Sci. USA 94, 13,317–13,322.
Haberle, W., Horber, J. K., and Binnig, G. (1991) Force microscopy on living cells. J. Vac. Sci. Technol. B 9, 1210–1213.
Horber, J. K., Haberle, W., Ohnesorge, F., Binnig, G., Liebich, H. G., Czerny, C. P., et al. (1992) Investigation of living cells in the nanometer regime with the scanning force microscope. Scan. Microsc. 6, 919–929.
Henderson, E. (1994) Imaging of living cells by atomic force microscopy. Prog. Surface Sci. 46, 39–60.
Henderson, R. M., Schneider, S., Li, Q., Hornby, D., White, S. J., and Oberleithner, H. (1996) Imaging ROMK1 inwardly rectifying ATP-sensitive K+ channel protein using atomic force microscopy. Proc. Natl. Acad. Sci. USA 93, 8756–8760.
Radmacher, M. (1997) Measuring the elastic properties of biological samples with the AFM. IEEE Eng. Med. Biol. 16, 47–57.
Schneider, S. W., Yano, Y., Sumpio, B. E., Jena, B. P., Geibel, J. P., Gekle, M., and Oberleithner, H. (1997) Rapid aldosterone-induced cell volume increase of endothelial cells measured by the atomic force microscope. Cell Biol. Int. 21, 759–768.
Fritzsche, W., Takac, L., and Henderson, E. (1997) Application of atomic force microscopy to visualization of DNA, chromatin, and chromosomes. [Review] [65 refs]. Crit. Rev. Eukaryotic Gene Exp. 7, 231–240.
Hansma, H. G., Kim, K. J., Laney, D. E., Garcia, R. A., Argaman, M., Allen, M. J., Parsonsand, S. M. (1997) Properties of biomolecules measured from atomic force microscope images: a review. [Review] [59 refs]. J. Struct. Biol. 119, 99–108.
Yang, Y., Sweeney, W. V., Schneider, K., Chait, B. T., and Tam, J. P. (1994) Two-step selective formation of three disulfide bridges in the synthesis of the C-terminal epidermal growth factor-like domain in human blood coagulation factor IX. Protein Sci. 3, 1267–1275.
Lal, R. and Yu, L. (1993) Atomic force microscopy of cloned nicotinic acetylcholine receptor expressed in Xenopus oocytes. Proc. Natl. Acad. Sci. USA 90, 7280–7284.
Lal, R. and John, S. A. (1994) Biological applications of atomic force microscopy. [Review]. Am. J. Physiol. 266, C1–21.
John, S. A., Saner, D., Pitts, J. D., Holzenburg, A., Finbow, M. E., and Lal, R. (1997) Atomic force microscopy of arthropod gap junctions. J. Struct. Biol. 120, 22–31.
Lal, R. (1996) Imaging molecular structure of channels and receptors with an atomic force microscope. Scan. Microsc. 10(Suppl.), 81–95.
Lal, R., John, S. A., Laird, D. W., and Arnsdorf, M. F. (1995) Heart gap junction preparations reveal hemiplaques by atomic force microscopy. Am. J. Physiol. 268, C968–C977.
Hoh, J. H., Sosinsky, G. E., Revel, J. P., and Hansma, P. K. (1993) Structure of the extracellular surface of the gap junction by atomic force microscopy. Biophys. J. 65, 149–163.
Oberleithner, H., Brinckmann, E., Schwab, A., and Krohne, G. (1994) Imaging nuclear pores of aldosterone-sensitive kidney cells by atomic force microscopy. Proc. Natl. Acad. Sci. USA 91, 9784–9788.
Folprecht, G., Schneider, S., and Oberleithner, H. (1996) Aldosterone activates the nuclear pore transporter in cultured kidney cells imaged with atomic force microscopy. Pflugers Arch. Eur. J. Physiol. 432, 831–838.
Schneider, S., Folprecht, G., Krohne, G., and Oberleithner, H. (1995) Immunolocalization of lamins and nuclear pore complex proteins by atomic force microscopy. Pflugers Arch. Eur. J. Physiol. 430, 795–801.
Perez-Terzic, C., Pyle, J., Jaconi, M., Stehno-Bittel, L., and Clapham, D. E. (1996) Conformational states of the nuclear pore complex induced by depletion of nuclear Ca2+ stores. Science 273, 1875–1877.
Wang, H. and Clapham, D. E. (1999) Conformational changes of the in situ nuclear pore complex. Biophys. J. 77, 241–247.
Radmacher, M., Fritz, M., and Hansma, P. K. (1995) Imaging soft samples with the atomic force microscope: gelatin in water and propanol. Biophys. J. 69, 264–270.
Wagner, P. (1998) Immobilization strategies for biological scanning probe microscopy. [Review] [58 refs]. FEBS Lett. 430, 112–115.
Korchev, Y. E., Bashford, C. L., Milovanovic, M., Vodyanoy, I., and Laband, M. J. (1997) Scanning ion conductance microscopy of living cells. Biophys. J. 73, 653–658.
Merkel, R., Nassoy, P., Leung, A., Ritchie, K., and Evans, E. (1999) Energy landscapes of receptor-ligand bonds explored with dynamic force spectroscopy. Nature 397, 50–53.
Engel, A., Lyubchenko, Y., and Muller, D. (1999) Atomic force microscopy: a powerful tool to observe biomolecules at work. [Review] [39 refs]. Trends Cell Biol. 9, 77–80.
Schabert, F. A., Henn, C., and Engel, A. (1995) Native Escherichia coli OmpF porin surfaces probed by atomic force microscopy. Science 268, 92–94.
Muller, D. J. and Engel, A. (1999) Voltage and pH-induced channel closure of porin OmpF visualized by atomic force microscopy. J. Mol. Biol. 285, 1347–1351.
Muller, D. J., Sass, H. J., Muller, S. A., Buldt, G., and Engel, A. (1999) Surface structures of native bacteriorhodopsin depend on the molecular packing arrangement in the membrane. J. Mol. Biol. 285, 1903–1909.
Yang, J., Mou, J., and Shao, Z. (1994) Structure and stability of pertussis toxin studied by in situ atomic force microscopy. FEBS Lett. 338, 89–92.
Czajkowsky, D. M. and Shao, Z. (1998) Submolecular resolution of single macromolecules with atomic force microscopy. [Review] [40 refs]. FEBS Lett. 430, 51–54.
Fisher, T. E., Marszalek, P. E., Oberhauser, A. F., Carrion, V., and Fernandez, J. M. (1999) The micro-mechanics of single molecules studied with atomic force microscopy. [Review] [35 refs]. J. Physiol. 520 Pt 1, 5–14.
Shao, Z., Yang, J., and Somlyo, A. P. (1995) Biological atomic force microscopy: from microns to nanometers and beyond. [Review] [134 refs]. Annu. Rev. Cell. Dev. Biol. 11, 241–265.
Fritz, M., Radmacher, M., and Gaub, H. E. (1994) Granula motion and membrane spreading during activation of human platelets imaged by atomic force microscopy. Biophys. J. 66, 1328–1334.
Muller, D. J., Baumeister, W., and Engel, A. (1996) Conformational change of the hexagonally packed intermediate layer of Deinococcus radiodurans monitored by atomic force microscopy. J. Bacteriol. 178, 3025–3030.
Thalhammer, S., Stark, R. W., Muller, S., Wienberg, J., and Heckl, W. M. (1997) The atomic force microscope as a new microdissecting tool for the generation of genetic probes. J. Struct. Biol. 119, 232–237.
Fotiadis, D., Muller, D. J., Tsiotis, G., Hasler, L., Tittmann, P., Mini, T., et al. (1998) Surface analysis of the photosystem I complex by electron and atomic force microscopy. J. Mol. Biol. 283, 83–94.
Muller, D. J., Buldt, G., and Engel, A. (1995) Force-induced conformational change of bacteriorhodopsin. J. Mol. Biol. 249, 239–243.
Schoenenberger, C. A. and Hoh, J. H. (1994) Slow cellular dynamics in MDCK and R5 cells monitored by time-lapse atomic force microscopy. Biophys. J. 67, 929–936.
Radmacher, M., Fritz, M., Kacher, C. M., Cleveland, J. P., and Hansma, P. K. (1996) Measuring the viscoelastic properties of human platelets with the atomic force microscope. Biophys. J. 70, 556–567.
Rief, M., Gautel, M., Oesterhelt, F., Fernandez, J. M., and Gaub, H. E. (1997) Reversible unfolding of individual titin immunoglobulin domains by AFM [see comments]. Science 276, 1109–1112.
Florin, E. L., Rief, M., Lehmann, H., Ludwig, M., Dornmair, C., Moy, V. T., and Gaub, H. E. (1995) Sensing specific molecular interasctions with the Atomic Force Microscope. Biosens. Bioelectron. 10, 895–901.
Moy, V. T., Florin, E. L., and Gaub, H. E. (1994) Adhesive forces between ligand and receptor measured by Afm. Colloids Surfaces A-Physicochem. Eng. Aspects 93, 343–348.
Bustamante, J. O., Liepins, A., Prendergast, R. A., Hanover, J. A., and Oberleithner, H. (1995) Patch clamp and atomic force microscopy demonstrate TATA-binding protein (TBP) interactions with the nuclear pore complex. J. Membr. Biol. 146, 263–272.
Horber, J. K., Mosbacher, J., and Haberle, W. (1995) Force microscopy on membrane patches, in Single-Channel Recording (Sakmann, B. and Neher, E., eds.), Plenum, New York, pp. 375–393.
Hoh, J. H. and Schoenenberger, C. A. (1994) Surface morphology and mechanical properties of MDCK monolayers by atomic force microscopy. J. Cell Sci. 107, 1105–1114.
Sokabe, M. and Sachs, F. (1990) The structure and dynamics of patch-clamped membranes: a study by differential interference microscopy. J. Cell Biol. 111, 599–606.
McEwen, B. F., Song, M. J., Ruknudin, A., Barnard, D. P., Frank, J., and Sachs, F. (1990) Tomographic three dimensional reconstruction of patch clamped membranes imaged with the high voltage electron microscope. XII Intl. Cong. El. Microsc. 522–523 (Abstract).
Ruknudin, A., Song, M. J., and Sachs, F. (1991) The ultrastructure of patchclamped membranes: a study using high voltage electron microscopy. J. Cell Biol. 112, 125–134.
Akinlaja, J. and Sachs, F. (1998) The breakdown of cell membranes by electrical and mechanical stress. Biophys. J. 75, 247–254.
Horber, J. K. H., Mosbacher, J., Haberle, W., Ruppersberg, J. P., and Sakmann, B. (1995) A look at membrane patches with a scanning force microscope. Biophys. J. 68, 1687–1693.
Mosbacher, J., Haberle, W., and Horber, J. K. (1996) Studying membranes with scanning force microscopy and patch-clamp. J. Vac. Sci. Technol. B 14, 1449–1452.
Larmer, J., Schneider, S. W., Danker, T., Schwab, A., and Oberleithner, H. (1997) Imaging excised apical plasma membrane patches of MDCK cells in physiological conditions with atomic force microscopy. Pflugers Arch. Eur. J. Physiol. 434, 254–260.
Crowe, J. H. and Crowe, L. M. (1982) Induction of anhydrobiosis: membrane changes during drying. Cryobiology 19, 317–328.
Danker, T., Mazzanti, M., Tonini, R., Rakowska, A., and Oberleithner, H. (1997) Using atomic force microscopy to investigate patch-clamped nuclear membrane. Cell Biol. Int. 21, 747–757.
Bustamante, J. O. (1994) Nuclear electrophysiology. J. Membr. Biol. 138, 105–112.
Mosbacher, J., Langer, M., Horber, H., and Sachs, F. (1998) Voltage-dependent membrane displacements measure by atomic force microscopy. J. Gen. Physiol. 111, 65–74.
Petrov, A. G. (1999) Lyotropic State of Matter: Molecular Physics and Living Matter Physics. Gordon & Breach Publishing Group, Amsterdam.
Todorov, A. T., Petrov, A. G., and Fendler, J. H. (1994) 1st observation of the converse flexoelectric effect in bilayer-lipid membranes. J. Phys. Chem. 98, 3076–3079.
Todorov, A. T., Petrov, A. G., and Fendler, J. H. (1994) Flexoelectricity of charged and polar bilayer-lipid membranes studied by stroboscopic interferometry. Langmuir 10, 2344–2350.
Petrov, A. G., Ramsey, R. L., and Usherwood, P. N. (1989) Curvature-electric effects in artificial and natural membranes studied using patch-clamp techniques. Eur. Biophys. J. 17, 13–17.
Petrov, A. G., Miller, B. A., Hristova, K., and Usherwood, P. N. (1993) Flexoelectric effects in model and native membranes containing ion channels. Eur. Biophys. J. 22, 289–300.
Iwamoto, H., Czajkowsky, D. M., Cover, T. L., Szabo, G., and Shao, Z. (1999) VacA from Helicobacter pylori: a hexameric chloride channel. FEBS Lett. 450, 101–104.
Bett, G. C. L. and Sachs, F. (2000) Activation and inactivation of mechanosensitive currents in the chick heart. J. Membr. Biol. 173, 237–254.
Langer, M. G., Offner, W., Wittmann, H., Flosser, H., Schaar, H., Haberle, W., et al. (1997) A scanning force microscope for simultaneous force and patchclamp measurements on living cell tissues. Rev. Sci. Instr. 68, 2583–2590.
Snyder, K., Besch, S. D., Zhang, P. C., and Sachs, F. (2000) Hardware and software modifications of the Quesant AFM for use in biology. In press.
Sachs, F. (1995) A low drift micropipette holder. Eur. J. Physiol. 429, 434–435.
Snyder, K. V., Kreigstein, A. and Sachs, F. (1999) A convenient electrode holder for glass pipettes to stabilize electrode potentials. Eur. J. Physiol. 438, 405–411.
Beyder, A., Snyder, K. V., and Sachs, F. (2000) New AFM cantilever for low force applications in liquids. In press.
Ashmore, J. F. (1987) A fast motile response in guinea-pig outer hair cells: the cellular basis of the cochlear amplifier. J. Physiol. (Lond.) 388, 323–347.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2001 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Snyder, K., Zhang, P.C., Sachs, F. (2001). Dynamic AFM of Patch Clamped Membranes. In: Lopatin, A.N., Nichols, C.G. (eds) Ion Channel Localization. Methods in Pharmacology and Toxicology. Humana Press. https://doi.org/10.1385/1-59259-118-3:425
Download citation
DOI: https://doi.org/10.1385/1-59259-118-3:425
Publisher Name: Humana Press
Print ISBN: 978-0-89603-833-2
Online ISBN: 978-1-59259-118-3
eBook Packages: Springer Protocols