Skip to main content
Biophysical Journal logoLink to Biophysical Journal
. 2001 Oct;81(4):2226–2240. doi: 10.1016/S0006-3495(01)75870-5

Anomalous subdiffusion in fluorescence photobleaching recovery: a Monte Carlo study.

M J Saxton 1
PMCID: PMC1301694  PMID: 11566793

Abstract

Anomalous subdiffusion is hindered diffusion in which the mean-square displacement of a diffusing particle is proportional to some power of time less than one. Anomalous subdiffusion has been observed for a variety of lipids and proteins in the plasma membranes of a variety of cells. Fluorescence photobleaching recovery experiments with anomalous subdiffusion are simulated to see how to analyze the data. It is useful to fit the recovery curve with both the usual recovery equation and the anomalous one, and to judge the goodness of fit on log-log plots. The simulations show that the simplest approximate treatment of anomalous subdiffusion usually gives good results. Three models of anomalous subdiffusion are considered: obstruction, fractional Brownian motion, and the continuous-time random walk. The models differ significantly in their behavior at short times and in their noise level. For obstructed diffusion the approach to the percolation threshold is marked by a large increase in noise, a broadening of the distribution of diffusion coefficients and anomalous subdiffusion exponents, and the expected abrupt decrease in the mobile fraction. The extreme fluctuations in the recovery curves at and near the percolation threshold result from extreme fluctuations in the geometry of the percolation cluster.

Full Text

The Full Text of this article is available as a PDF (1.0 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Almeida P. F., Vaz W. L., Thompson T. E. Lateral diffusion and percolation in two-phase, two-component lipid bilayers. Topology of the solid-phase domains in-plane and across the lipid bilayer. Biochemistry. 1992 Aug 11;31(31):7198–7210. doi: 10.1021/bi00146a024. [DOI] [PubMed] [Google Scholar]
  2. Almeida P. F., Vaz W. L., Thompson T. E. Lateral diffusion in the liquid phases of dimyristoylphosphatidylcholine/cholesterol lipid bilayers: a free volume analysis. Biochemistry. 1992 Jul 28;31(29):6739–6747. doi: 10.1021/bi00144a013. [DOI] [PubMed] [Google Scholar]
  3. Almeida P. F., Vaz W. L., Thompson T. E. Percolation and diffusion in three-component lipid bilayers: effect of cholesterol on an equimolar mixture of two phosphatidylcholines. Biophys J. 1993 Feb;64(2):399–412. doi: 10.1016/S0006-3495(93)81381-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Axelrod D., Koppel D. E., Schlessinger J., Elson E., Webb W. W. Mobility measurement by analysis of fluorescence photobleaching recovery kinetics. Biophys J. 1976 Sep;16(9):1055–1069. doi: 10.1016/S0006-3495(76)85755-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brown D. A., London E. Structure and function of sphingolipid- and cholesterol-rich membrane rafts. J Biol Chem. 2000 Jun 9;275(23):17221–17224. doi: 10.1074/jbc.R000005200. [DOI] [PubMed] [Google Scholar]
  6. Cherry R. J., Smith P. R., Morrison I. E., Fernandez N. Mobility of cell surface receptors: a re-evaluation. FEBS Lett. 1998 Jun 23;430(1-2):88–91. doi: 10.1016/s0014-5793(98)00595-x. [DOI] [PubMed] [Google Scholar]
  7. Coelho F. P., Vaz W. L., Melo E. Phase topology and percolation in two-component lipid bilayers: a monte Carlo approach. Biophys J. 1997 Apr;72(4):1501–1511. doi: 10.1016/S0006-3495(97)78798-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Edidin M. Lipid microdomains in cell surface membranes. Curr Opin Struct Biol. 1997 Aug;7(4):528–532. doi: 10.1016/s0959-440x(97)80117-0. [DOI] [PubMed] [Google Scholar]
  9. Feder T. J., Brust-Mascher I., Slattery J. P., Baird B., Webb W. W. Constrained diffusion or immobile fraction on cell surfaces: a new interpretation. Biophys J. 1996 Jun;70(6):2767–2773. doi: 10.1016/S0006-3495(96)79846-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Ghosh R. N., Webb W. W. Automated detection and tracking of individual and clustered cell surface low density lipoprotein receptor molecules. Biophys J. 1994 May;66(5):1301–1318. doi: 10.1016/S0006-3495(94)80939-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gordon G. W., Chazotte B., Wang X. F., Herman B. Analysis of simulated and experimental fluorescence recovery after photobleaching. Data for two diffusing components. Biophys J. 1995 Mar;68(3):766–778. doi: 10.1016/S0006-3495(95)80251-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Jacobson K., Dietrich C. Looking at lipid rafts? Trends Cell Biol. 1999 Mar;9(3):87–91. doi: 10.1016/s0962-8924(98)01495-0. [DOI] [PubMed] [Google Scholar]
  13. Kusumi A., Sako Y. Cell surface organization by the membrane skeleton. Curr Opin Cell Biol. 1996 Aug;8(4):566–574. doi: 10.1016/s0955-0674(96)80036-6. [DOI] [PubMed] [Google Scholar]
  14. London E., Brown D. A. Insolubility of lipids in triton X-100: physical origin and relationship to sphingolipid/cholesterol membrane domains (rafts). Biochim Biophys Acta. 2000 Nov 23;1508(1-2):182–195. doi: 10.1016/s0304-4157(00)00007-1. [DOI] [PubMed] [Google Scholar]
  15. Lopez A., Dupou L., Altibelli A., Trotard J., Tocanne J. F. Fluorescence recovery after photobleaching (FRAP) experiments under conditions of uniform disk illumination. Critical comparison of analytical solutions, and a new mathematical method for calculation of diffusion coefficient D. Biophys J. 1988 Jun;53(6):963–970. doi: 10.1016/S0006-3495(88)83177-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Munnelly H. M., Roess D. A., Wade W. F., Barisas B. G. Interferometric fringe fluorescence photobleaching recovery interrogates entire cell surfaces. Biophys J. 1998 Aug;75(2):1131–1138. doi: 10.1016/S0006-3495(98)77602-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Nagle J. F. Long tail kinetics in biophysics? Biophys J. 1992 Aug;63(2):366–370. doi: 10.1016/S0006-3495(92)81602-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Rietveld A., Simons K. The differential miscibility of lipids as the basis for the formation of functional membrane rafts. Biochim Biophys Acta. 1998 Nov 10;1376(3):467–479. doi: 10.1016/s0304-4157(98)00019-7. [DOI] [PubMed] [Google Scholar]
  19. Saxton M. J. Anomalous diffusion due to binding: a Monte Carlo study. Biophys J. 1996 Mar;70(3):1250–1262. doi: 10.1016/S0006-3495(96)79682-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Saxton M. J. Anomalous diffusion due to obstacles: a Monte Carlo study. Biophys J. 1994 Feb;66(2 Pt 1):394–401. doi: 10.1016/s0006-3495(94)80789-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Saxton M. J., Jacobson K. Single-particle tracking: applications to membrane dynamics. Annu Rev Biophys Biomol Struct. 1997;26:373–399. doi: 10.1146/annurev.biophys.26.1.373. [DOI] [PubMed] [Google Scholar]
  22. Saxton M. J. Single-particle tracking: the distribution of diffusion coefficients. Biophys J. 1997 Apr;72(4):1744–1753. doi: 10.1016/S0006-3495(97)78820-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Schram V., Lin H. N., Thompson T. E. Topology of gel-phase domains and lipid mixing properties in phase-separated two-component phosphatidylcholine bilayers. Biophys J. 1996 Oct;71(4):1811–1822. doi: 10.1016/S0006-3495(96)79382-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Schram V., Tocanne J. F., Lopez A. Influence of obstacles on lipid lateral diffusion: computer simulation of FRAP experiments and application to proteoliposomes and biomembranes. Eur Biophys J. 1994;23(5):337–348. doi: 10.1007/BF00188657. [DOI] [PubMed] [Google Scholar]
  25. Schwille P., Haupts U., Maiti S., Webb W. W. Molecular dynamics in living cells observed by fluorescence correlation spectroscopy with one- and two-photon excitation. Biophys J. 1999 Oct;77(4):2251–2265. doi: 10.1016/S0006-3495(99)77065-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Schwille P., Korlach J., Webb W. W. Fluorescence correlation spectroscopy with single-molecule sensitivity on cell and model membranes. Cytometry. 1999 Jul 1;36(3):176–182. doi: 10.1002/(sici)1097-0320(19990701)36:3<176::aid-cyto5>3.0.co;2-f. [DOI] [PubMed] [Google Scholar]
  27. Sheets E. D., Lee G. M., Simson R., Jacobson K. Transient confinement of a glycosylphosphatidylinositol-anchored protein in the plasma membrane. Biochemistry. 1997 Oct 14;36(41):12449–12458. doi: 10.1021/bi9710939. [DOI] [PubMed] [Google Scholar]
  28. Simons K., Ikonen E. How cells handle cholesterol. Science. 2000 Dec 1;290(5497):1721–1726. doi: 10.1126/science.290.5497.1721. [DOI] [PubMed] [Google Scholar]
  29. Simons K., Toomre D. Lipid rafts and signal transduction. Nat Rev Mol Cell Biol. 2000 Oct;1(1):31–39. doi: 10.1038/35036052. [DOI] [PubMed] [Google Scholar]
  30. Simson R., Yang B., Moore S. E., Doherty P., Walsh F. S., Jacobson K. A. Structural mosaicism on the submicron scale in the plasma membrane. Biophys J. 1998 Jan;74(1):297–308. doi: 10.1016/S0006-3495(98)77787-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Smith P. R., Morrison I. E., Wilson K. M., Fernández N., Cherry R. J. Anomalous diffusion of major histocompatibility complex class I molecules on HeLa cells determined by single particle tracking. Biophys J. 1999 Jun;76(6):3331–3344. doi: 10.1016/S0006-3495(99)77486-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Soumpasis D. M. Theoretical analysis of fluorescence photobleaching recovery experiments. Biophys J. 1983 Jan;41(1):95–97. doi: 10.1016/S0006-3495(83)84410-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Vaz W. L., Melo E. C., Thompson T. E. Translational diffusion and fluid domain connectivity in a two-component, two-phase phospholipid bilayer. Biophys J. 1989 Nov;56(5):869–876. doi: 10.1016/S0006-3495(89)82733-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Wolf D. E. Designing, building, and using a fluorescence recovery after photobleaching instrument. Methods Cell Biol. 1989;30:271–306. doi: 10.1016/s0091-679x(08)60983-8. [DOI] [PubMed] [Google Scholar]

Articles from Biophysical Journal are provided here courtesy of The Biophysical Society

RESOURCES