Abstract
The specific organization and distribution of protein receptors and lipids on the cellular plasma membrane play a crucial role for the spatiotemporal control of many different cellular processes. A great deal of novel knowledge in this area is currently being generated thanks to the advent of modern surperresolution optical techniques combined with single-molecule approaches. In this chapter, we focus on near-field nanoscopy, a technique particularly well suited for the study of biological cell surfaces at the nanometer scale. We first describe the general concept of near-field scanning optical microscopy (NSOM) and specifically focus on how NSOM is being exploited to map the spatiotemporal organization of proteins and lipids. Novel routes toward surperresolution using optical nanoantennas and first applications for cell membrane nanoimaging are discussed. The last part of the chapter describes recent technical breakthroughs that enable the application of NSOM in living cells providing detailed dynamic information on diffusion processes occurring at the nanoscale.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Bibliography
Jares-Erijman EA, Jovin TM (2003) FRET imaging. Nat Biotechnol 21(11):1387–1395
Varma R, Mayor S (1998) GPI-anchored proteins are organized in submicron domains at the cell surface. Nature 394:798–801
Sharma P, Varma R, Sarasij RC, Ira GK, Krishnamoorthy G, Rao M, Mayor S (2004) Nanoscale organization of multiple GPI-anchored proteins in living cell membranes. Cell 116:577–589
Mayor S, Rao M (2004) Rafts: scale-dependent, active lipid organization at the cell surface. Traffic 5:231–240
Betzig E, Patterson GH, Sougrat R, Lindwasser OW, Olenych S, Bonifacino JS, Davidson MW, Lippincott-Schwartz J, Hess HF (2006) Imaging intracellular fluorescent proteins at nanometer resolution. Science 313:1642–1645
Hess ST, Girirajan TPK, Mason MD (2006) Ultra-high resolution imaging by fluorescence photoactivation localization microscopy. Biophys J 91:4258–4272
Rust MJ, Bates M, Zhuang X (2006) Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM). Nat Methods 3:793–795
Lillemeier BF, Mörtelmaier MA, Forstner MB, Huppa JB, Groves JT, Davis MM (2010) TCR and Lat are expressed on separate protein islands on T cell membranes and concatenate during activation. Nat Immunol 11:90–96
Sherman E, Barr V, Manley S, Patterson GH, Balagopalan L, Akpan I, Regan CK, Merrill RK, Sommers CL, Lippincott-Schwartz J, Samelson LE (2011) Functional nanoscale organization of signaling molecules downstream of the T cell antigen receptor. Immunity 35(5):705–720
Sengupta P, Jovanovic-Talisman T, Skoko D, Renz M, Veatch SL, Lippincott-Schwartz J (2011) Probing protein heterogeneity in the plasma membrane using PALM and pair correlation analysis. Nat Methods 8(11):969–975
Williamson DJ, Owen DM, Rossy J, Magenau A, Wehrmann M, Gooding JJ, Gaus K (2011) Pre-existing clusters of the adaptor Lat do not participate in early T cell signaling events. Nat Immunol 12:655–662
Sieber JJ, Willig KI, Kutzner C, Gerding-Reimers C, Harke B, Donnert G, Rammner B, Eggeling C, Hell SW, Grubmüller H, Lang T (2007) Anatomy and dynamics of a supramolecular membrane protein cluster. Science 317:1072–1076
Kellner RR, Baier CJ, Willig KI, Hell SW, Barrantes FJ (2007) Nanoscale organization of nicotinic acetylcholine receptors revealed by stimulated emission depletion microscopy. Neuroscience 144:135–143
Eggeling C, Ringemann C, Medda R, Schwarzmann G, Sandhoff K, Polyakova S, Belov VN, Hein B, von Middendorff C, Schönle A, Hell SW (2009) Direct observation of the nanoscale dynamics of membrane lipids in a living cell. Nature 457:1159–1162
Synge EA (1928) A suggested method for extending microscopic resolution into the ultra-microscopic region. Philos Mag 6:356–362.
Ash EA, Nicholls G (1972) Super-resolution aperture scanning microscope. Nature 237(5357):510–512
Pohl DW, Denk W, Lanz M (1984) Optical stethoscopy – image recording with resolution lambda/20. Appl Phys Lett 44(7):651–653
Lewis A, Isaacson M, Harootunian A, Muray A (1984) Development of a 500-a spatial-resolution light-microscope. 1. Light is efficiently transmitted through gamma-16 diameter apertures. Ultramicroscopy 13(3):227–231
Peasler MA, Moyer PJ (1996) Near-field optics: theory, instrumentation and applications. Wiley, New York
Betzig E, Trautman JK, Harris TD, Weiner JS, Kostelak RL (1991) Breaking the diffraction barrier: optical microscopy on a nanometric scale. Science 251:1468–1470
Stockle R, Fokas C, Deckert V, Zenobi R, Sick B, Hecht B, Wild UP (1999) High-quality near-field optical probes by tube etching. Appl Phys Lett 75(2):160–162
Burgos P, Lu Z, Ianoul A, Hnatovsky C, Viriot M-L, Johnston LJ, Taylor RS (2003) Near-field scanning optical microscopy probes: a comparison of pulled and double-etched bent NSOM probes for fluorescence imaging of biological samples. J Microscopy 211:37–47
Veerman JA, Otter AM, Kuipers L, van Hulst NF (1998) High definition aperture probes for near-field optical microscopy fabricated by focused ion beam milling. Appl Phys Lett 72:3115–3117
Betzig E, Finn PL, Weiner JS (1992) Combined shear force and near-field scanning optical microscopy. Appl Phys Lett 60:2484–2486
Toledo-Crow R, Yang PC, Chen Y, Vaez-Iravani M (1992) Near-field differential scanning optical microscope with atomic force regulation. Appl Phys Lett 60:2957–2959
Lieberman K, BenAmi N, Lewis A (1996) Fully integrated near-field optical, far-field optical, and normal-force scanned probe microscope. Rev Sci Instrum 67(10):3567–3572
Koopman M, de Bakker BI, Garcia-Parajo MF, van Hulst NF (2003) Shear force imaging of soft samples in liquid using a diving bell concept. Appl Phys Lett 83:5083–5085
Taylor RS, Vobornik D, Lu Z, Chisholm RA, Johnston LJ (2010) Damping behavior of bent fiber NSOM probes in water. J Appl Phys 107(4):043526
Höppener C, Novotny L (2008) Antenna-based optical imaging of single Ca2+ transmembrane proteins in liquids. Nano Lett 8:642–646
Höppener C, Siebrasse JP, Peters R, Kubitscheck U, Naber A (2005) High-resolution near-field optical imaging of single nuclear pore complexes under physiological conditions. Biophys J 88:3681–3688
Heinzelmann H, Pohl DW (1994) Scanning near-field optical microscopy. Appl Phys A 59(2):89–101
Garcia-Parajo MF, de Bakker BI, Koopman M, Cambi A, de Lange F, Figdor CG, van Hulst NF (2005) Near-field fluorescence microscopy: an optical nanotool to study protein organization at the cell membrane. Nanobiotechnology 1:113–120
de Lange F, Cambi A, Huijbens R, de Bakker BI, Rensen WHJ, Garcia-Parajo MF, van Hulst NF, Figdor CG (2001) Cell biology beyond the diffraction limit: near-field scanning optical microscopy. J Cell Sci 114:4153–4160
Koopman M, Cambi A, de Bakker BI, Joosten B, Figdor CG, van Hulst NF, Garcia-Parajo MF (2004) Near-field scanning optical microscopy in liquid for high resolution single molecule detection on dendritic cells. FEBS Lett 573:6–10
Singer SJ, Nicolson GL (1972) The fluid mosaic model of the structure of cell membranes. Science 175(4023):720–731
Jacobson K, Sheets ED, Simson R (1995) Revisiting the fluid mosaic model of membranes. Science 268(5216):1441–1442
Maxfield FR (2002) Plasma membrane microdomains. Curr Opin Cell Biol 14:483–487
Simons M, Friedrichson T, Schulz JB, Pitto M, Masserini M, Kurzchalia TV (1999) Exogenous administration of gangliosides displaces GPI-anchored proteins from lipid microdomains in living cells. Mol Biol Cell 10(10):3187–3196
Kusumi A, Suzuki KGN (2005) Toward understanding the dynamics of membrane-raft-based molecular interactions. Biochim Biophys Acta 1746(3):234–251
Douglass AD, Vale RD (2005) Single-molecule microscopy reveals plasma membrane microdomains created by protein-protein networks that exclude or trap signaling molecules in T cells. Cell 121:937–950
Lingwood D, Simons K (2010) Lipid rafts as a membrane-organizing principle. Science 327:46–50
Cambi A, Joosten B, Koopman M, de Lange F, Beeren I, Torensma R, Fransen JA, Garcia-Parajo MF, van Leeuwen FN, Figdor CG (2006) Organization of the integrin LFA-1 in nanoclusters regulates its activity. Mol Biol Cell 17:4270–4281
Cambi A, de Lange F, van Maarseveen NM, Nijhuis M, Joosten B, van Dijk EMHP, de Bakker BI, Fransen JA, Bovee-Geurts PHM, van Leeuwen FN, van Hulst NF, Figdor CG (2004) Microdomains of the C-type lectin DC-SIGN are portals for virus entry into dendritic cells. J Cell Biol 164:145–155
Mayor S, Pagano RE (2007) Pathways of clathrin-independent endocytosis. Nat Rev Mol Cell Biol 8:603–612
Mañes S, Viola A (2006) Lipid rafts in lymphocyte activation and migration. Mol Membr Biol 23:59–69
Suzuki KGN, Fujiwara TK, Sanematsu F, Iino R, Edidin MA, Kusumi A (2007) GPI-anchored receptor clusters transiently recruit Lyn and Gα for temporary cluster immobilization and Lyn activation: single-molecule tracking study 1. J Cell Biol 177:717–730
Lasserre R, Guo X-J, Conchonaud F, Hamon Y, Hawchar O, Bernard A-M, Soudja SMH, Lenne P-F, Rigneault H, Olive D, Bismuth G, Nunès JA, Payrastre B, Marguet D, He H-T (2008) Raft nanodomains contribute to Akt/PKB plasma membrane recruitment and activation. Nat Chem Biol 4:538–547
Prior IA, Muncke C, Parton RG, Hancock JF (2003) Direct visualization of Ras proteins in spatially distinct cell surface microdomains. J Cell Biol 160:165–170
Anderson RGW, Jacobson K (2002) A role for lipid shells in targeting proteins to caveolae, rafts, and other lipid domains. Science 296(5574):1821–1825
Cherukuri A, Carter RH, Brooks S, Bornmann W, Finn R, Dowd CS, Pierce SK (2004) B cell signaling is regulated by induced palmitoylation of CD81. J Biol Chem 279:31973–31982
Hemler ME (2005) Tetraspanin functions and associated microdomains. Nat Rev Mol Cell Biol 6:801–811
Maecker HT, Todd SC, Levy S (1997) The tetraspanin superfamily: molecular facilitators. FASEB J 11:428–442
Berditchevski F (2001) Complexes of tetraspanins with integrins: more than meets the eye. J Cell Sci 114:4143–4151
Charrin S, le Naour F, Silvie O, Milhiet P-E, Boucheix C, Rubinstein E (2009) Lateral organization of membrane proteins: tetraspanins spin their web. Biochem J 420:133–154
Yanez-Mo M, Barreiro O, Gordon-Alonso M, Sala-Valdés M, Sanchez-Madrid F (2009) Tetraspanin-enriched microdomains: a functional unit in cell plasma membranes. Trends Cell Biol 19:434–446
Barreiro O, Zamai M, Yanez-Mo M, Tejera E, López-Romero P, Monk PN, Gratton E, Caiolfa VR, Sanchez-Madrid F (2008) Endothelial adhesion receptors are recruited to adherent leukocytes by inclusion in preformed tetraspanin nanoplatforms. J Cell Biol 183:527–542
Belanis L, Plowman SJ, Rotblat B, Hancock JF, Kloog Y (2008) Galectin-1 is a novel structural component and a major regulator of H-ras nanoclusters. Mol Biol Cell 19(4):1404–1414
Nieminen J, Kuno A, Hirabayashi J, Sato S (2007) Visualization of galectin-3 oligomerization on the surface of neutrophils and endothelial cells using fluorescence resonance energy transfer. J Biol Chem 282(2):1374–1383
Kusumi A, Sako Y (1996) Cell surface organization by the membrane skeleton. Curr Opin Cell Biol 8:566–574
Kusumi A, Nakada C, Ritchie K, Murase K, Suzuki KGN, Murakoshi H, Kasai RS, Kondo J, Fujiwara TK (2005) Paradigm shift of the plasma membrane concept from the two-dimensional continuum fluid to the partitioned fluid: high-speed single-molecule tracking of membrane molecules. Annu Rev Biophys Biomol Struct 34:351–378
Andrews NL, Lidke KA, Pfeiffer JR, Burns AR, Wilson BS, Oliver JM, Lidke DS (2008) Actin restricts FcεRI diffusion and facilitates antigen-induced receptor immobilization. Nat Cell Biol 10(8):955–963
Betzig E, Trautman JK (1992) Near-field optics: microscopy, spectroscopy, and surface modification beyond the diffraction limit. Science 257:189–195
Enderle T, Ha T, Ogletree DF, Chemla DS, Magowan C, Weiss S (1997) Membrane specific mapping and colocalization of malarial and host skeletal proteins in the Plasmodium falciparum infected erythrocyte by dual-color near-field scanning optical microscopy. Proc Natl Acad Sci 94(2):520–525
Chen Y, Qin J, Chen Z W (2008) Fluorescence-topographic NSOM directly visualizes peak-valley polarities of GM1/GM3 rafts in cell membrane fluctuations. J Lipid Res 49(10):2268–2275
Gomez-Mouton C, Abad JL, Mira E, Lacalle RA, Gallardo E, Jimenez-Baranda S, Illa I, Bernad A, Mañes S, Martinez-A C (2001) Segregation of leading-edge and uropod components into specific lipid rafts during T cell polarization. Proc Natl Acad Sci 98(17):9642–9647
Abulrob A, Lu Z, Brunette E, Pulla D, Stanimirovic D, Johnston LJ (2008) Near-field scanning optical microscopy detects nanoscale glycolipid domains in the plasma membrane. J Microscopy 232:225–234
van Zanten TS, Gómez J, Manzo C, Cambi A, Buceta J, Reigada R, Garcia-Parajo MF (2010) Direct mapping of nanoscale compositional connectivity on intact cell membranes. Proc Natl Acad Sci 107:15437–15442
Lillemeier BF, Pfeiffer JR, Surviladze Z, Wilson BS, Davis MM (2006) Plasma membrane-associated proteins are clustered into islands attached to the cytoskeleton. Proc Natl Acad Sci 103:18992–18997
de Bakker BI, de Lange F, Cambi A, Korterik JP, van Dijk EMHP, van Hulst NF, Figdor CG, Garcia-Parajo MF (2007) Nanoscale organization of the pathogen receptor DC-SIGN mapped by single-molecule high-resolution fluorescence microscopy. ChemPhysChem 8:1473–1480
de Bakker BI, Bodná A, van Dijk EMHP, Vámosi G, Damjanovich S, Waldmann TA, van Hulst NF, Jenei A, Garcia-Parajo MF (2008) Nanometer-scale organization of the alpha subunits of the receptors for IL2 and IL15 in human T lymphoma cells. J Cell Sci 121:627–633.
Vobornik D, Rouleau Y, Haley J, Bani-Yaghoub M, Taylor RS, Johnston LJ, Pezacki JP (2009) Nanoscale organization of β2-adrenergic receptor-Venus fusion protein domains on the surface of mammalian cells. Biochem Biophys Res Commun 382:85–90
Abulrob A, Lu Z, Baumann E, Vobornik D, Taylor RS, Stanimirovic D, Johnston LJ (2010) Nanoscale imaging of epidermal growth factor receptor clustering: effects of inhibitors. J Biol Chem 285:3145–3156
Chen J, Pei Y, Chen ZW, Cai J (2010) Quantum dot labeling based on near-field optical imaging of CD44 molecules. Micron 41(3):198–202
Zhong L, Zhang Z, Lu X, Huang D, Chen CY, Wang RC, Chen ZW (2011) NSOM/QD-based fluorescence and topographic image fusion directly reveals nano-spatial peak and valley polarities of CD69 and CD71 activation molecules on cell-membrane fluctuations during T-cell activation. Immunol Lett 140(1–2):44–51
Zhong L, Zeng G, Lu X, Wang RC, Gong G, Yan L, Huang D, Chen ZW (2009) NSOM/QD-based direct visualization of CD3-induced and CD28-enhanced nanospatial coclustering of TCR and coreceptor in nanodomains in T cell activation. PloS one 4:e5945
Ianoul A, Grant DD, Rouleau Y, Bani-Yaghoub M, Johnston LJ, Pezacki JP (2005) Imaging nanometer domains of beta-adrenergic receptor complexes on the surface of cardiac myocytes. Nat Chem Biol 1:196–202
van Zanten TS, Cambi A, Koopman M, Joosten B, Figdor CG, Garcia-Parajo MF (2009) Hotspots of GPI-anchored proteins and integrin nanoclusters function as nucleation sites for cell adhesion. Proc Natl Acad Sci 106:18557–18562
Meder D, Moreno MJ, Verkade P, Vaz WLC, Simons K (2006) Phase coexistence and connectivity in the apical membrane of polarized epithelial cells. Proc Natl Acad Sci 103:329–334
Kenworthy AK, Nichols BJ, Remmert CL, Hendrix GM, Kumar M, Zimmerberg J, Lippincott-Schwartz J (2004) Dynamics of putative raft-associated proteins at the cell surface. J Cell Biol 165:735–746
Saxton MJ, Jacobson K (1997) Single-particle tracking: applications to membrane dynamics. Annu Rev Biophys Biomol Struct 26:373–399
Schmidt T, Schütz GJ, Baumgartner W, Gruber HJ, Schindler H (1996) Imaging of single molecule diffusion. Proc Natl Acad Sci 93:2926–2929
Schwille P, Haupts U, Maiti S, Webb WW (1999) Molecular dynamics in living cells observed by fluorescence correlation spectroscopy with one- and two-photon excitation. Biophys J 77(4):2251–2265
Kim SA, Heinze KG, Schwille P (2007) Fluorescence correlation spectroscopy in living cells. Nat Methods 4:963–973
Banks DS, Fradin C (2005) Anomalous diffusion of proteins due to molecular crowding. Biophys J 89:2960–2971
Vobornik D, Banks DS, Lu Z, Fradin C, Taylor RS, Johnston LJ (2008) Fluorescence correlation spectroscopy with sub-diffraction-limited resolution using near-field optical probes. Appl Phys Lett 93:163904
Herrmann M, Neuberth N, Wissler J, Pérez J, Gradl D, Naber A (2009) Near-field optical study of protein transport kinetics at a single nuclear pore. Nano Lett 9:3330–3336
Manzo C, van Zanten TS, Garcia-Parajo MF (2011) Nanoscale fluorescence correlation spectroscopy on intact living cell membranes with NSOM probes. Biophys J 100:L8–L10
Schwille P, Haustein E (2009) Fluorescence correlation spectroscopy an introduction to its concepts and applications. Anal Chem 94(22):1–33
Sahl SJ, Leutenegger M, Hilbert M, Hell SW, Eggeling C (2010) Fast molecular tracking maps nanoscale dynamics of plasma membrane lipids. Proc Natl Acad Sci 107:6829–6834
Wawrezinieck L, Rigneault H, Marguet D, Lenne P-F (2005) Fluorescence correlation spectroscopy diffusion laws to probe submicron membrane organization. Biophys J 89:4029–4042
Wenger J, Conchonaud F, Dintinger J, Wawrezinieck L, Ebbesen TW, Rigneault H, Marguet D, Lenne P-F (2007) Diffusion analysis within single nanometric apertures reveals the ultrafine cell membrane organization. Biophys J 92:913–919
Neumann L, Pang Y, Houyou A, Juan ML, Gordon R, van Hulst NF (2011) Extraordinary optical transmission brightens near-field fiber probe. Nano Lett 11:355–360
Taminiau TH, Stefani FD, Segerink FB, van Hulst NF (2008) Optical antennas direct single-molecule emission. Nat Photonics 2:234–237
Kinkhabwala A, Yu Z, Fan S, Avlasevich Y, Müllen K, Moerner WE (2009) Large single-molecule fluorescence enhancements produced by a bowtie nanoantenna. Nat Photonics 3:654–657
Höppener C, Beams R, Novotny L (2009) Background suppression in near-field optical imaging. Nano Lett 9:903–908
Frey HG, Witt S, Felderer K, Guckenberger R (2004) High-resolution imaging of single fluorescent molecules with the optical near-field of a metal tip. Phys Rev Lett 93:200801
Taminiau TH, Moerland RJ, Segerink FB, Kuipers L, van Hulst NF (2007) λ/4 resonance of an optical monopole antenna probed by single molecule fluorescence. Nano Lett 7:28–33
Betzig E, Chichester RJ (1993) Single molecules observed by near-field scanning optical microscopy. Science 262:1422–1425
Bethe HA (1944) Theory of diffraction by small holes. Phys Rev 66:163–182
Bouwkamp CJ (1950) On Bethe’s theory of diffraction by small holes. Philips Res Rep 5:321–332
Garcia-Parajo MF (2008) Optical antennas focus in on biology. Nat Photonics 2:201–203
Novotny L (2007) Effective wavelength scaling for optical antennas. Phys Rev Lett 98:266802
Thompson RE, Larson DR, Webb WW (2002) Precise nanometer localization analysis for individual fluorescent probes. Biophys J 82:2775–2783
van Zanten TS, Lopez-Bosque MJ, Garcia-Parajo MF (2010) Imaging individual proteins and nanodomains on intact cell membranes with a probe-based optical antenna. Small 6:270–275
Hrelescu C, Sau TK, Rogach AL, Jäckel F, Laurent G, Douillard L, Charra F (2011) Selective excitation of individual plasmonic hotspots at the tips of single gold nanostars. Nano Lett 11:402–407
Mivelle M, Ibrahim IA, Baida F, Burr GW, Nedeljkovic D, Charraut D, Rauch JY, Salut R, Grosjean T (2010) Bowtie nano-aperture as interface between near-fields and a single-mode fiber. Opt Express 18(15):15964–15974
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer-Verlag Berlin Heidelberg
About this chapter
Cite this chapter
van Zanten, T.S., Manzo, C., Garcia-Parajo, M.F. (2012). Near-Field Optical Nanoscopy of Biological Membranes. In: Mély, Y., Duportail, G. (eds) Fluorescent Methods to Study Biological Membranes. Springer Series on Fluorescence, vol 13. Springer, Berlin, Heidelberg. https://doi.org/10.1007/4243_2012_52
Download citation
DOI: https://doi.org/10.1007/4243_2012_52
Published:
Publisher Name: Springer, Berlin, Heidelberg
Print ISBN: 978-3-642-33127-5
Online ISBN: 978-3-642-33128-2
eBook Packages: Chemistry and Materials ScienceChemistry and Material Science (R0)