Skip to main content

Near-Field Scanning Optical Microscopy to Identify Membrane Microdomains

  • Protocol

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 400))

Abstract

Near-field scanning optical microscopy (NSOM) allows optical imaging with a spatial resolution that is significantly better than the diffraction limited resolution achievable with conventional optical microscopy. NSOM has the potential to study the nanoscale organization of membrane surfaces and ultimately to resolve questions concerning lipid rafts in both model-supported membranes and cellular membranes. Supported phospholipid monolayers and bilayers of phase separated binary and ternary lipid mixtures that model the composition of lipid rafts in natural membranes have been studied by NSOM. The results illustrate the ability of NSOM measurements with 50–100 nm probe apertures to obtain detailed nanoscale information for small, closely spaced domains, and the utility of two-color experiments to probe localization of raft markers in supported membranes.

This is a preview of subscription content, log in via an institution.

Buying options

Protocol
USD   49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   129.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   169.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD   169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Binder, W. H., Barragan, V., and Menger, F. M. (2003) Domains and rafts in lipid membranes. Angew. Chem. Int. Ed. 42, 5802–5827.

    Article  CAS  Google Scholar 

  2. Simons, K. and Vaz, W. L. C. (2004) Model systems, lipid rafts and cell membranes. Annu. Rev. Biophys. Biomol. Struct. 33, 269–295.

    Article  PubMed  CAS  Google Scholar 

  3. London, E. (2002) Insights into lipid raft structure and formation from experiments in model membranes. Curr. Opin. Struct. Biol. 12, 480–486.

    Article  PubMed  CAS  Google Scholar 

  4. Simons, K. and Ikonen, E. (1997) Functional rafts in cell membranes. Nature 387, 589–572.

    Article  Google Scholar 

  5. Anderson, R. G. W. and Jacobson, K. (2002) A role for lipid shells in targeting proteins to cave-olae, rafts and other lipid domains. Science 296, 1821–1825.

    Article  PubMed  CAS  Google Scholar 

  6. Pike, L. J. (2004) Lipid rafts: heterogeneity on the high seas. Biochem. J. 378, 281–292.

    Article  PubMed  CAS  Google Scholar 

  7. Munro, S. (2003) Lipid rafts: elusive or illusive? Cell 115, 377–388.

    Article  PubMed  CAS  Google Scholar 

  8. Dunn, R. C. (1999) Near-field scanning optical microscopy. Chem. Rev. 99, 2891–2927.

    Article  PubMed  CAS  Google Scholar 

  9. Edidin, M. (2001) Near-field scanning optical microscopy, a siren call to biology. Traffic 2, 797–803.

    Article  PubMed  CAS  Google Scholar 

  10. Lewis, A., Taha, H., Strinkovski, A., et al. (2003) Near-field optics: from subwavelength illumination to nanometric shadowing. Nat. Biotechnol. 21, 1378–1386.

    Article  PubMed  CAS  Google Scholar 

  11. Ianoul, A., Street, M., Grant, D., Pezacki, J., Taylor, R. S., and Johnston, L. J. (2004) Near-field scanning fluorescence microscopy study of ion channel clusters in cardiac myocyte membranes. Biophys. J. 87, 3525–3535.

    Article  PubMed  CAS  Google Scholar 

  12. Ianoul, A., Grant, D., Rouleau, Y., Bani, 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.

    Article  PubMed  CAS  Google Scholar 

  13. Burgos, P., Lu, Z., Ianoul, A., et al. (2003) Near-field scanning optical microscopy probes: a comparison of pulled and double-etched bent NSOM probes for fluorescence imaging of biological samples. J. Micros. 211, 37–47.

    Article  CAS  Google Scholar 

  14. Burgos, P., Yuan, C., Viriot, M.-L., and Johnston, L. J. (2003) Two-color near-field fluorescence microscopy studies of microdomains (‘rafts’) in model membranes. Langmuir 19, 8002–8009.

    Article  CAS  Google Scholar 

  15. Ianoul, A., Burgos, P., Lu, Z., Taylor, R. S., and Johnston, L. J. (2003) Phase separation in complex supported phospholipid bilayers visualized by near-field scanning optical microscopy in liquid. Langmuir 19, 9246–9254.

    Article  CAS  Google Scholar 

  16. Murray, J., Cuccia, L., Ianoul, A., Cheetham, J. J., and Johnston, L. J. (2004) Imaging the selective binding of synapsin to anionic membrane domains. Chem. Biol. Chem. 5, 1489–1494.

    CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2007 Humana Press Inc.

About this protocol

Cite this protocol

Ianoul, A., Johnston, L.J. (2007). Near-Field Scanning Optical Microscopy to Identify Membrane Microdomains. In: Dopico, A.M. (eds) Methods in Membrane Lipids. Methods in Molecular Biology™, vol 400. Humana Press. https://doi.org/10.1007/978-1-59745-519-0_31

Download citation

  • DOI: https://doi.org/10.1007/978-1-59745-519-0_31

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-662-7

  • Online ISBN: 978-1-59745-519-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics