Abstract
We use submicrometer apertures milled in an aluminium film to study the diffusion dynamics of β-Bodipy-FL-C5-HPC (Bodipy-PC) fluorophores in a lipid dioleoylphosphatidylcholine (DOPC) multilayer. The observation volume is limited by the aperture diameter, well below the optical wavelength. This spatial resolution improvement comes together with an enhancement of the detected fluorescence per molecule as compared to an open sample, with a significant increase up to 3.5 times.
Key words: fluorescence correlation spectroscopy, confocal microscopy, nano-apertures, lipid bilayer
References
- 1.Degiron A., et al. Optical Transmission Properties of a Single Subwavelength Aperture in a Real Metal Opt. Com. 200423961–66.2004OptCo.239...61D [Google Scholar]
- 2.Levene M.J., et al. Zero-Mode Waveguides for Single-Molecule Analysis at High Concentrations Science 2003299682–686. 10.1126/science.10797002003Sci...299..682L [DOI] [PubMed] [Google Scholar]
- 3.Samiee K.T., Foquet M., Guo L., Cox E.G., Craighead H.G. Repressor Oligomerization Kinetics at High Concentrations Using Fluorescence Correlation Spectroscopy in Zero-Mode Waveguides. Biophys J. 2005;88:2145–2153. doi: 10.1529/biophysj.104.052795. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 4.Rigneault H., et al. Enhancement of Single-Molecule Fluorescence Detection in Subwavelength Apertures Phys. Rev. Lett. 200595117401–117404. 10.1103/PhysRevLett.95.1174012005PhRvL..95k7401R [DOI] [PubMed] [Google Scholar]
- 5.Edel J.B., Wu M., Baird B., Craighead H.G. High Spatial Resolution Observation of Single Molecule Dynamics in Living Cell Membranes. Biophys. J. 2005;88:L43–L45. doi: 10.1529/biophysj.105.061937. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 6.Magde D., Elson E., Webb W.W.Thermodynamic Fluctuations in a Reacting System Measurement by Fluorescence Correlation Spectroscopy Phys. Rev. Lett. 197229705–708. 10.1103/PhysRevLett.29.7051972PhRvL..29..705M [DOI] [Google Scholar]
- 7.Singer S.J., Nicolson G.L.The Fluid Mosaic Model Science 1972175720–731.1972Sci...175..720S [DOI] [PubMed] [Google Scholar]
- 8.Samiee, K.T., et al.: Zero-Mode Waveguides for Single-Molecule Spectroscopy on Lipid Membranes, Biophys. J. doi: 10.1529/biophysj.105.072819. [DOI] [PMC free article] [PubMed]