Abstract
Cells respond to biochemical and mechanical stimuli through a series of steps that begin at the molecular, nanometre level, and translate finally in global cell response. Defects in biochemical- and/or mechanical-sensing, transduction or cellular response are the cause of multiple diseases, including cancer and immune disorders among others. Within the booming field of regenerative medicine, there is an increasing need for developing and applying nanotechnology tools to bring understanding on the cellular machinery and molecular interactions at the nanoscale. Nanotechnology, nanophotonics and in particular, high-resolution-based fluorescence approaches are already delivering crucial information on the way that cells respond to their environment and how they organize their receptors to perform specialized functions. This chapter focuses on emerging super-resolution optical techniques, summarizing their principles, technical implementation, and reviewing some of the achievements reached so far.
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
Stephens, D. J., and Allan, V. J. (2003) Light microscopy techniques for live cell imaging. Science 300, 82–86.
Michalet, X., Kapanidis, A. N., Laurence, T., Pinaud, F., Doose, S., Pfughoefft, M., and Weiss, S. (2003) The power and prospects of fluorescence microscopies and spectroscopies. Annu. Rev. Biophys. Biomol. Struct. 32, 161–182.
Betzig, E., and Chichester, R. J. (1993) Single molecules observed by near-field scanning optical microscopy. Science 262, 1422–1425.
Weiss, S. (1999) Fluorescence spectroscopy of single biomolecules. Science 283, 1676–1683.
Tsien, R. Y. (1998) The green fluorescent protein Annu. Rev. Biochem. 67, 509–544.
Lippincott-Schwartz, J., and Patterson, G. H. (2003) Development and use of fluorescence protein markers in living cells. Science 300, 87–91.
Hansma, P. K., Cleveland, J. P., Radmacher, M., Walters, D. A., Hillner, P. E., Bezanilla, M., Fritz, M., Vie, D., Hansma, H. G., Prater, C. B. (1994) Tapping mode atomic force microscopy in liquids Appl. Phys. Lett. 64, 1738–1740.
Putman, C. A., van der Werf, K. O., de Groot, B. G., van Hulst, N. F., and Greve, J. (1994) Tapping mode atomic force microscopy in liquid Appl. Phys. Lett. 64, 2454–2456.
Willemsen, O. H., Snel, M. M., Cambi, A., Gerve, J., de Groot, B. G., and Figdor, C. G. (2000) Biomolecular interactions measured by atomic force microscopy Biophys. J. 79, 3267–3281.
Stroh, C., Wang, H., Bash, R., Ashcroft, B., Nelson, J., Gruber, H., Lohr, D., Lindsay, S. M., and Hinterdorfer, P (2004) Single-molecule recognition imaging microscopy Proc. Natl. Acad. Sci. USA 101, 12503–12507.
Donnert, G., Keller, J., Medda, R., Andrei, M. A., Rizzoli, S. O., Luhrmann, R., Jahn, R., Eggeling, C., and Hell, S. W. (2006) Macromolecular-scale resolution in biological fluorescence microscopy Proc. Natl. Acad. Sci. USA 103, 11440–11445.
Willig, K. I., Rizzoli, S. O., Wesphal, V., Jahn, R., and Hell, S. W. (2006) STED microscopy reveals hat synaptotagmin remains clustered after synpatic vesicle exocytosis Nature 440, 935–939.
Sieber, J. J., Willig, K. I., Kutzner, C., Gerding-Reimers, C., Harke, B., Donnert, G., Rammner, B., Eggeling, C., Hell, S. W., Grubmuller, H., and Lang, T. (2007) Anatomy and dynamics of a supramolecular membrane protein cluster Science 317, 1072–1076.
Kellner, R. R., Baier, C. J., Willig, K. I., Hell, S. W., and Barrantes, F. J. (2007) Nanoscale organization of nicotinic acetylcholine receptors revealed by stimulated emission depletion microscopy Neuroscience 144, 135–143.
Wesphal, V., Rizzoli, S. O., Lauterbach, M. A., Kamin, D., Jahn, R., and Hell, S. W. (2008) Video-rate far-field optical nanoscopy dissects synaptic vesicle movement Science 320, 246–249.
Gustafsson, M. G. L. (2005) Non-linear structured-illumination microscopy: wide-field fluorescence imaging with theoretically unlimited resolution Proc. Natl. Acad. Sci. USA 102, 13081–13086.
Schermelleh, L., Carlton, P. M., Haase, S., Shao, L., Winoto, L., Kner, P., Burke, B., Cardoso, M. C., Agard, D. A., Gustafsson, M. G. L., Leonhardt, H., and Sedat, J. W. (2008) Subdiffraction multicolor imaging of the nuclear periphery with 3D structured illumination microscopy Science 320, 1332–1336.
Gustaffson, M. G. L., Shao, L., Carlton, P. M., Wang, C. J. R., Golubovskaya, I. N., Cande, W. Z., Agard, D. A., and Sedat, J. W. (2008) Three-dimensional resolution doubling in wide-field fluorescence microscopy by structured illumination Biophys. J. 94, 4957–4970.
Pielage, J., Cheng, L., Fetter, R. D., Carlton, P. M., Sedat, J. W., and Davis, G. W. (2008) A presynaptic giant ankrin stabilizes the NMJ through regulation of presynaptics microtubules and transsynaptic cell adhesion Neuron 58, 195–209.
Betzig, E., Patterson, G. H., Sougrat, R., Lindwasser, O. W. S., Olenuch, Bonifacino, J. S., Davidson, M. W., Lippincott-Schwartz, J., and Hess, H. F. (2006) Imaging intracellular fluorescent proteins at nanometer resolution Science 313, 1642–1645.
Hess, S. T., Girirajan, T. P. K., and Mason, M. D. (2006) Ultra-high resolution imaging by fluorescence photoactivation localization microscopy Biophys. J. 91, 4258–4272.
Rust, M. J., Bates, M., and Zhuang, X. (2006) Subdiffraction limit imaging by stochastic optical reconstruction microscopy (STORM) Nat. Methods 3, 793–796.
Schroff, H., Galbraith, C. G., Galbraith, J. A., White, H., Gillete, J., Olenych, S., Davidson, M. W., and Betzig, E. (2007) Dual-color superresolution imaging of genetically expressed probes within adhesion complexes Proc. Natl. Acad. Sci. USA 104, 20308–20313.
Manley, S., Gillete, J. M., Patterson, G. H., Schroff, H., Hess, H. F., Betzig, E., and Lippincott-Schwartz, J. (2008) High-density mapping of single molecule trajectories with photoactivated localization microscopy Nat. Methods 5, 155–157.
Hess, S. T., Gould, T. J., Gudheti, M. V., Maas, S. A., Mills, K. D., and Zimmerberg, J. (2007) Dynamic clustered distribution of hemagglutinin resolved at 40nm in living cell membranes discriminates between raft theories Proc. Natl. Acad. Sci. USA 104, 17370–17375.
Schroff, H., Galbraith, C. G., Galbraith, J. A., and Betzig, E. (2008) Live cell photoactivated localization microscopy of nanoscale adhesion dynamics Nat. Methods 5, 417–423.
Pohl, D. W., Denk, W., and Lanz, M. (1984) Optical stethoscopy: image recording with resolution l/20 Appl. Phys. Lett. 44, 651–653.
Betzig, E., Trautman, J. K., Harris, T. D., Weiner, J. S., Kostelak, R. L. (1991) Breaking the diffraction barrier: optical microscopy on a nanometric scale Science 251, 1468–1470.
van Hulst, N. F., Garcia-Parajo, M. F., Moers, M. H. P., Veerman, J. A., and Ruiter, A. G. T. (1997) Near-field fluorescence imaging of genetic material: towards the molecular limit J. Struct. Bio. 119, 222–231.
Hecht, B., Sick, B., Wild, U. P., Deckert, V., Zenobi, R., Martin O. J. F. and Dieter, D. W. (2000) Scanning near-field optical microscopy with aperture probes: fundamentals and applications J. Chem. Phys. 112, 7761–7774.
Karrai, K., and Grober, R. D. (1995) Piezo-electric tip-sample distance control for near field optical microscopes Appl. Phys. Lett. 66, 1842–1844.
Ruiter, A. G., Veerman, J. A., van der Werf, K. O., van Hulst, N. F. (1997) Dynamic behavior of tuning fork shear-force feedback Appl. Phys. Lett. 71, 28–30.
Zenhausern, F., Martin, Y., and Wickramasinghe, H. K. (1995) Scanning interferometric apertureless microscopy: Optical imaging at 10 Angstrom resolution Science 269, 1083–1085.
Novotny, L., and Stranick, S. J. (2006) Near-field optical microscopy and spectroscopy with pointed probes Annu. Rev. Phys. Chem. 57, 303–331.
Sanchez, E. J., Novotny, L., Sunney-Xie, X. (1999) Near-field fluorescence microscopy based on two-photon excitation with metal tips Phys. Rev. Lett. 82, 4014–4017.
Anger, P., Bharadwaj, P., and Novotny, L. (2006) Enhancement and quenching of single-molecule fluorescence Phys. Rev. Lett. 96, 1130021–1130024.
Höppener, C., and Novotny, L. (2008) Antenna-based optical imaging of single Ca2+ transmembrane proteins in liquids Nano Lett. 8, 642–646.
Höppener, C., Beams, R., Novotny, L. (2009) Background suppression in near-field optical imaging Nano Lett. 9, 903–908.
Koopman, M., de Bakker B. I., Garcia-Parajo, M. F., and van Hulst, N. F. (2003) Shear force imaging of soft samples in liquid using a diving bell concept Appl. Phys. Lett. 83, 5083–5085.
Koopman, M., Cambi, A., de Bakker, B. I., Joosten, B., Figdor, C. G., van Hulst, N. F., and Garcia-Parajo, M.F. (2004) Near-field scanning optical microscopy in liquid for high resolution single molecule detection on dendritic cells FEBS Lett. 573, 6–10.
Höppener, C., Siebrasse, J. P., Peters, R., Kubitscheck, U., and Naber, A. (2005) High-resolution near-field optical imaging of single nuclear pore complexes under physiological conditions Biophys. J. 88, 3681–3688.
Korchev, Y. E., Raval, M., Lab, M. J., Gorelik, J., Edwards, C. R. W., Rayment, T., and Klenerman, D. (2000) Hybrid scanning ion conductance and scanning near-field optical microscopy for the study of living cells Biophys. J. 78, 2675–2679.
de Bakker, B. I., de Lange, F., Cambi, A., Korterik, J. P., van Dijk, E. M. H. P., van Hulst, N. F., Figdor, C. G., and Garcia-Parajo, M. F. (2007) Nanoscale organization of the pathogen receptor DC-SIGN mapped by single-molecule high-resolution fluorescence microscopy Chemphyschem 8, 1473–1480.
Chen, Y., Shao, L., Ali, Z., Cai, J., and Chen, Z. W. (2008) NSOM/QD-based nanoscale immunofluorescence imaging of antigen-specific T-cell receptor responses during an in vivo clonal Vγ2Vδ2 T-cell expansion Blood 111, 4220–4232.
Enderle, T., Ha, T., Ogletree, D. F., Chemla, D. S., Magowan, C., and 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. USA 94, 520–525.
de Bakker, B. I., Bodnar, A., van Dijk, E. P., Vámosi, G., Damjanovich, S., Waldmann, T. A., van Hulst, N. F., Jenei A., and Garcia-Parajo, M. F. (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.
Ianoul, A., Grant, D. D., Rouleau, Y., Bani-Yaghoub, M., Johnston, L. J., and Pezacki, J. P. (2005) Imaging nanometer domains of β-adrenergic receptor complexes on the surface of cardiac myocytes Nat. Chem. Biol. 1, 196–202.
van Zanten, T. S., Cambi, A., Koopman, M., Joosten, B., Figdor, C. G., and Garcia-Parajo, M. F. (2009) Hotspots of GPI-anchored proteins and integrin nanoclusters function as nucleation sites for cell adhesion Proc. Natl. Acad. Sci. USA 106, 18557–18562.
Vobornik, D., Banks, D. S., Lu, Z., Fradin, C., Taylor, R., and Johnston, L. J. (2008) Fluorescence correlation spectroscopy with sub-diffraction-limited resolution using near-field optical probes Appl. Phys. Lett. 93, 163904–163906.
Garcia-Parajo, M. F. (2008) Optical antennas focus in on biology Nat. Photonics 2, 201–203.
Frey, H. G., 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, 200801–200804.
Taminiau, T. H., Moerland, R. J., Segerink, F. B., Kuipers, L. van Hulst, N. F. (2007) λ/4 Resonance of an optical monopole antenna probed by single molecule fluorescence Nano Lett. 7, 28–33.
van Zanten, T. S., Lopez, M. J., Garcia-Parajo, M. F. (2009) Imaging individual proteins and nanodomains on intact cell membranes with a probe-based optical antenna Small 6, 270–275.
Denk, W., Strickler, J. H., and Webb, W. W. (1990) Two-photon laser scanning fluorescente microscopy Science 248, 73–76.
Miller, M. J., Wei, S. H., Cahalan, M. D., and Parker, I. (2003) Autonomous T cell trafficking examined in vivo with intravital two-photon microscopy Proc. Nat. Acad. Sci. USA 100, 2604–2609.
Bousso, P., and Robey, E. A. (2004) Dynamic behavior of T cells and thymocytes in lymphoid organs as revealed by two-photon microscopy Immunity 21, 349–355.
Nimmerjahn, A., Kirchhoff, F., and Helmchen, F. (2005) Resting microglial cells are highly dynamic surveillants of brain parenchyma Science 308, 1314–1318.
Davalos, D. et al. (2005) ATP mediates rapid microglial response to local brain injury in vivo Nat. Neurosci 8, 752–758.
Helmchen, F., and Denk, W. (2005) Deep tissue two-photon microscopy Nat. Methods 2, 932–940.
Jain, R. K., Munn, L. L., and Fukumura, D. (2002) Dissecting tumor pathophysiology using infravital microscopy Nat. Rev. Cancer 2, 266–276.
Skoch, J., Hickey, G. A., Kajadasz, S. T., Hyman, B. T., and Bacskai, B. J. (2005) In-vivo imaging of amyloid-beta deposits in mouse brain with multiphoton microscopy Methods Mol. Bio. 299, 349–363.
Tsai, J., Grutzendler, J., Duff, K., and Gan, W. (2004) Fibrillar amyloid deposition leads to local synaptic abnormalities and breakage of neuronal branches Nat. Neurosci. 7, 1181–1183.
Bacskai, B. J. et al. (2003) Four-dimensional multiphoton imaging of brain entry, amyloid binding, and clearance of an amyloid-beta ligand in transgenic mice Proc. Natl. Acad. Sci. USA 100, 12462–12467.
Acknowledgments
The author would like to thank B. I. de Bakker, M. Koopman and T.S. van Zanten for NSOM images and fruitful discussions. Financial support has been provided by the EC-RTN-IMMUNANOMAP and EC-NEST-BIO-LIGHT-TOUCH, Spanish Ministry of Science and Technology MAT2007-66629-C02-01 and Generalitat de Catalunya 2009 SGR 597.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Garcia-Parajo, M.F. (2012). The Role of Nanophotonics in Regenerative Medicine. In: Navarro, M., Planell, J. (eds) Nanotechnology in Regenerative Medicine. Methods in Molecular Biology, vol 811. Humana Press. https://doi.org/10.1007/978-1-61779-388-2_17
Download citation
DOI: https://doi.org/10.1007/978-1-61779-388-2_17
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61779-387-5
Online ISBN: 978-1-61779-388-2
eBook Packages: Springer Protocols