Skip to main content
SpringerLink
Log in

Advances in Image Correlation Spectroscopy: Measuring Number Densities, Aggregation States, and Dynamics of Fluorescently labeled Macromolecules in Cells

  • Review Paper
  • Published:
Cell Biochemistry and Biophysics Aims and scope Submit manuscript

Abstract

A brief historical outline of fluorescence fluctuation correlation techniques is presented, followed by an in-depth review of the theory and development of image correlation techniques, including: image correlation spectroscopy (ICS), temporal ICS (TICS), image cross-correlation spectroscopy (ICCS), spatiotemporal ICS (STICS), k-space ICS (kICS), raster ICS (RICS), and particle ICS (PICS). These techniques can be applied to analyze image series acquired on commercially available laser scanning or total internal reflection fluorescence microscopes, and are used to determine the number density, aggregation state, diffusion coefficient, velocity, and interaction fraction of fluorescently labeled molecules or particles. A comprehensive review of the application of ICS techniques to a number of systems, including cell adhesion, membrane receptor aggregation and dynamics, virus particle fusion, and fluorophore photophysics, is presented.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9
Fig. 10
Fig. 11

Similar content being viewed by others

References

  1. Heldin, C.-H. (1995). Dimerization of cell surface receptors in signal transduction. Cell, 80, 213–223.

    Article  PubMed  CAS  Google Scholar 

  2. Lemmon, M. A., & Schlessinger, J. (1994). Regulation of signal transduction and signal diversity by receptor oligomerization. Trends in Biochemical Sciences, 19, 459–463.

    Article  PubMed  CAS  Google Scholar 

  3. Sheets, E. D., Simson, R., & Jacobson, K. (1995). New insights into membrane dynamics from the analysis of cell surface interactions by physical methods. Current Opinion in Cell Biology, 7, 707–714.

    Article  PubMed  CAS  Google Scholar 

  4. Wiseman, P. W., Brown, C. M., Webb, D. J., Hebert, B., Johnson, N. L., Squier, J. A., Ellisman, M. H., & Horwitz, A. F. (2004). Spatial mapping of integrin interactions and dynamics during cell migration by image correlation microscopy. Journal of Cell Science, 117, 5521–5534.

    Article  PubMed  CAS  Google Scholar 

  5. Tserkovnyak, Y., & Nelson, D. R. (2006). Conditions for extreme sensitivity of protein diffusion in membranes to cell environments. Proceedings of the National Academy of Sciences of the United States of America, 103, 15002–15007.

    Article  PubMed  CAS  Google Scholar 

  6. Tsien, R. Y. (1998). The green fluorescent protein. Annual Review of Biochemistry, 67, 509–544.

    Article  PubMed  CAS  Google Scholar 

  7. Magde, D., Elson, E., & Webb, W. W. (1972). Thermodynamic fluctuations in a reacting system—measurement by fluorescence correlation spectroscopy. Physical Review Letters, 29, 705–708.

    Article  CAS  Google Scholar 

  8. Elson, E. L., & Magde, D. (1974). Fluorescence correlation spectroscopy. I. Conceptual basis and theory. Biopolymers, 13, 1–27.

    Article  CAS  Google Scholar 

  9. Magde, D., Elson, E. L., & Webb, W. W. (1974). Fluorescence correlation spectroscopy. II. An experimental realization. Biopolymers, 13, 29–61.

    Article  PubMed  CAS  Google Scholar 

  10. Magde, D., Webb, W. W., & Elson, E. L. (1978). Fluorescence correlation spectroscopy. III. Uniform translation and laminar flow. Biopolymers, 17, 361–376.

    Article  CAS  Google Scholar 

  11. Weissman, M., Schindler, H., & Feher, G. (1976). Determination of molecular weights by fluctuation spectroscopy: Application to DNA. Proceedings of the National Academy of Sciences of the United States of America, 73, 2776–2780.

    Article  PubMed  CAS  Google Scholar 

  12. Petersen, N. O. (1986). Scanning fluorescence correlation spectroscopy. I. Theory and simulation of aggregation measurements. Biophysical Journal, 49, 809–815.

    PubMed  CAS  Google Scholar 

  13. Petersen, N. O., Johnson, D. C., & Schlesinger, M. J. (1986). Scanning fluorescence correlation spectroscopy. II. Application to virus glycoprotein aggregation. Biophysical Journal, 49, 817–820.

    PubMed  CAS  Google Scholar 

  14. St-Pierre, P. R., & Petersen, N. O. (1990). Relative ligand binding to small or large aggregates measured by scanning correlation spectroscopy. Biophysical Journal, 58, 503–511.

    PubMed  CAS  Google Scholar 

  15. Meyer, T., & Schindler, H. (1988). Particle counting by fluorescence correlation spectroscopy. Simultaneous measurement of aggregation and diffusion of molecules in solutions and in membranes. Biophysical Journal, 54, 983–993.

    PubMed  CAS  Google Scholar 

  16. Koppel, D. E., Morgan, F., Cowan, A. E., & Carson, J. H. (1994). Scanning concentration correlation spectroscopy using the confocal laser microscope. Biophysical Journal, 66, 502–507.

    PubMed  CAS  Google Scholar 

  17. Petersen, N. O., Hoddelius, P. L., Wiseman, P. W., Seger, O., & Magnusson, K. E. (1993). Quantitation of membrane receptor distributions by image correlation spectroscopy: Concept and application. Biophysical Journal, 65, 1135–46.

    PubMed  CAS  Google Scholar 

  18. Berland, K. M., So, P. T., Chen, Y., Mantulin, W. W., & Gratton, E. (1996). Scanning two-photon fluctuation correlation spectroscopy: Particle counting measurements for detection of molecular aggregation. Biophysical Journal, 71, 410–420.

    PubMed  CAS  Google Scholar 

  19. Fink, M. C., Adair, K. V., Guenza, M. G., & Marcus, A. H. (2006). Translational diffusion of fluorescent proteins by molecular Fourier imaging correlation spectroscopy. Biophysical Journal, 91, 3482–3498.

    Article  PubMed  CAS  Google Scholar 

  20. Hebert, B., Costantino, S., & Wiseman, P. W. (2005). Spatiotemporal image correlation spectroscopy (STICS) theory, verification, and application to protein velocity mapping in living CHO cells. Biophysical Journal, 88, 3601–3614.

    Article  PubMed  CAS  Google Scholar 

  21. Cannell, M. B., McMorland, A., & Soeller, C. (2006). Image enhancement by deconvolution. In J. B. Pawley (Ed.), Handbook of biological confocal microscopy (pp. 488–500). New York: Springer.

    Google Scholar 

  22. Wiseman, P. W., & Petersen, N. O. (1999). Image correlation spectroscopy. II. Optimization for ultrasensitive detection of preexisting platelet-derived growth factor-beta receptor oligomers on intact cells. Biophysical Journal, 76, 963–77.

    PubMed  CAS  Google Scholar 

  23. Costantino, S., Comeau, J. W. D., Kolin, D. L., & Wiseman, P. W. (2005). Accuracy and dynamic range of spatial image correlation and cross-correlation spectroscopy. Biophysical Journal, 89, 1251–1260.

    Article  PubMed  CAS  Google Scholar 

  24. Wiseman, P. W., Höddelius, P., Petersen, N. O., & Magnusson, K. (1997). Aggregation of PDGF-β receptors in human skin fibroblasts: Characterization by image correlation spectroscopy (ICS). FEBS Letters, 401, 43–48.

    Article  PubMed  CAS  Google Scholar 

  25. Wiseman, P. W., Squier, J. A., Ellisman, M. H., & Wilson, K. R. (2000). Two-photon image correlation spectroscopy and image cross-correlation spectroscopy. Journal of Microscopy, 200, 14–25.

    Article  PubMed  CAS  Google Scholar 

  26. Yoo, H., Song, I., & Gweon, D.-G. (2006). Measurement and restoration of the point spread function of fluorescence confocal microscopy. Journal of Microscopy, 221, 172–176.

    Article  PubMed  CAS  Google Scholar 

  27. Huang, Z., & Thompson, N. L. (1996). Imaging fluorescence correlation spectroscopy: Nonuniform IgE distributions on planar membranes. Biophysical Journal, 70, 2001–2007.

    PubMed  CAS  Google Scholar 

  28. Rocheleau, J. V., Wiseman, P. W., & Petersen, N. O. (2003). Isolation of bright aggregate fluctuations in a multipopulation image correlation spectroscopy system using intensity subtraction. Biophysical Journal, 84, 4011–22.

    PubMed  CAS  Google Scholar 

  29. Wiseman, P. W., Capani, F., Squier, J. A., & Martone, M. E. (2002). Counting dendritic spines in brain tissue slices by image correlation spectroscopy analysis. Journal of Microscopy, 205, 177–86.

    Article  PubMed  CAS  Google Scholar 

  30. Palmer, A. G., & Thompson, N. L. (1989). High-order fluorescence fluctuation analysis of model protein clusters. Proceedings of the National Academy of Sciences of the United States of America, 86, 6148–6152.

    Article  PubMed  CAS  Google Scholar 

  31. Vanden Broek, W., Huang, Z., & Thompson, N. L. (1999). High-order autocorrelation with imaging fluorescence correlation spectroscopy: Application to IgE on supported planar membranes. Journal of Fluorescence, 9, 313–324.

    Article  CAS  Google Scholar 

  32. Sergeev, M., Costantino, S., & Wiseman, P. W. (2006). Measurement of monomer–oligomer distributions via fluorescence moment image analysis. Biophysical Journal, 91, 3884–3896.

    Article  PubMed  CAS  Google Scholar 

  33. Benn, A. G., & Kulperger, R. J. (1997). Integrated marked poisson processes with application to image correlation spectroscopy. The Canadian Journal of Statistics, 25, 215–231.

    Article  Google Scholar 

  34. Srivastava, M., & Petersen, N. O. (1996). Image cross-correlation spectroscopy: A new experimental biophysical approach to measurement of slow diffusion of fluorescent molecules. Methods in Cell Science, 18, 47–54.

    Article  Google Scholar 

  35. Srivastava, M., & Petersen, N. O. (1998). Diffusion of transferrin receptor clusters. Biophysical Chemistry, 75, 201–211.

    Article  PubMed  CAS  Google Scholar 

  36. Nohe, A., Keating, E., Fivaz, M., van der Goot, F. G., & Petersen, N. O. (2006). Dynamics of GPI-anchored proteins on the surface of living cells. Nanomedicine 39, 159–169.

    Google Scholar 

  37. Petersen, N. (2001). FCS and spatial correlations on biological surfaces. In R. Rigler & E. S. Elson (Eds.), Fluorescence correlation spectroscopy (pp. 162–184). Heidelberg: Springer.

    Google Scholar 

  38. Ulrich, M., Wohland, T., Rigler, R., & Vogel, H. (1999). Resolution of fluorescence correlation measurements. Biophysical Journal, 76, 1619–1631.

    Google Scholar 

  39. Feder, T. J., Brust-Mascher, I., Slattery, J. P., Baird, B., & Webb, W. W. (1996). Constrained diffusion or immobile fraction on cell surfaces: A new interpretation. Biophysical Journal, 70, 2767–2773.

    PubMed  CAS  Google Scholar 

  40. Aragón, S. R., & Pecora, R. (1976). Fluorescence correlation spectroscopy as a probe of molecular dynamics. The Journal of Chemical Physics, 64, 1791–1803.

    Article  Google Scholar 

  41. Saxton, M. J., & Jacobson, K. (1997). Single-particle tracking: Applications to membrane dynamics. Annual Review of Biophysics and Biomolecular Structure, 26, 373–399.

    Article  PubMed  CAS  Google Scholar 

  42. Kulkarni, R. P., Wu, D. D., Davis, M. E., & Fraser, S. E. (2005). Quantitating intracellular transport of polyplexes by spatio-temporal image correlation spectroscopy. Proceedings of the National Academy of Sciences of the United States of America, 102, 7523–7528.

    Article  PubMed  CAS  Google Scholar 

  43. Kolin, D. L., Costantino, S., & Wiseman, P. W. (2006). Sampling effects, noise, and photobleaching in temporal image correlation spectroscopy. Biophysical Journal, 90, 628–639.

    Article  PubMed  CAS  Google Scholar 

  44. Lagerholm, B. C., Weinreb, G. E., Jacobson, K., & Thompson, N. L. (2005). Detecting microdomains in intact cell membranes. Annual Review of Physical Chemistry, 56, 309–336.

    Article  PubMed  CAS  Google Scholar 

  45. Axelrod, D., Koppel, D. E., Schlessinger, J., Elson, E., & Webb, W. W. (1976). Mobility measurement by analysis of fluorescence photobleaching recovery kinetics. Biophysical Journal, 16, 1055–1069.

    PubMed  CAS  Google Scholar 

  46. Jacobson, K., O’Dell, D., & August, J. T. (1984). Lateral diffusion of an 80,000–dalton glycoprotein in the plasma membrane of murine fibroblasts: Relationships to cell structure and function. The Journal of Cell Biology, 99, 1624–1633.

    Article  PubMed  CAS  Google Scholar 

  47. Sheetz, M. P., & Koppel, D. E. (1979). Membrane damage caused by irradiation of fluorescent concanavalin. Proceedings of the National Academy of Sciences of the United States of America, 76, 3314–3317.

    Article  PubMed  CAS  Google Scholar 

  48. Lepock, J. R., Campbell, S. D., Gruber, M., & Kruuv, J. (1979). Photoinduced cell killing and crosslinking of fluorescein conjugated concanavalin A to cell surface proteins. Biochemical and Biophysical Research Communications, 91, 1157–1165.

    Article  PubMed  CAS  Google Scholar 

  49. Jacobson, K., Hou, Y., & Wojcieszyn, J. (1979). Evidence for lack of damage during photobleaching measurements of the lateral mobility of cell surface components. Experimental Cell Research, 116, 179–189.

    Article  Google Scholar 

  50. Wolf, D. E., Edidin, M., & Dragsten, P. R. (1980). Effect of bleaching light on measurements of lateral diffusion in cell membranes by the fluorescence photobleaching recovery method. Proceedings of the National Academy of Sciences of the United States of America, 77, 2043–2045.

    Article  PubMed  CAS  Google Scholar 

  51. Schwille, P., Korlach, J., & Webb, W. W. (1999). Fluorescence correlation spectroscopy with single-molecule sensitivity on cell and model membranes. Cytometry, 36, 176–182.

    Article  PubMed  CAS  Google Scholar 

  52. Nicolau, J., Dan V., Hancock, J. F., & Burrage, K. (2007). Sources of anomalous diffusion on cell membranes: A Monte Carlo study. Biophysical Journal, 92, 1975–1987.

    Article  PubMed  CAS  Google Scholar 

  53. Manders, E. M. M., Verbeek, F. J., & Aten, J. A. (1993). Measurement of colocalization of objects in dual-color confocal images. Journal of Microscopy, 169, 375–382.

    Google Scholar 

  54. Costes, S. V., Daelemans, D., Cho, E. H., Dobbin, Z., Pavlakis, G., & Lockett, S. (2004). Automatic and quantitative measurement of protein–protein colocalization in live cells. Biophysical Journal, 86, 3993–4003.

    Article  PubMed  CAS  Google Scholar 

  55. Comeau, J., Costantino, S., & Wiseman, P. W. (2006). A guide to accurate colocalization measurements. Biophysical Journal, 91, 4611–4622.

    Article  PubMed  CAS  Google Scholar 

  56. Manders, E. M. M. (1997). Chromatic shift in multicolour confocal microscopy. Journal of Microscopy, 185, 321–328.

    Article  Google Scholar 

  57. McAuliffe, M. J., Lalonde, F. M., McGarry, D., Gandler, W., Csaky, K., & Trus, B. L. (2001). Medical image processing, analysis & visualization in clinical research in 14th IEEE symposium on computer-based medical systems IEEE computer society (p. 0381).

  58. Schwille, P. (2001). Cross-correlation analysis. In R. Rigler & E. S. Elson (Eds.), FCS in fluorescence correlation spectroscopy: Theory and applications. New York: Springer. Volume 65 of Chemical Physics.

  59. Sekar, R. B., & Periasamy, A. (2003). Fluorescence resonance energy transfer (FRET) microscopy imaging of live cell protein localizations. The Journal of Cell Biology, 160, 629–633.

    Article  PubMed  CAS  Google Scholar 

  60. Milon, S., Hovius, R., Vogel, H., & Wohland, T. (2003). Factors influencing fluorescence correlation spectroscopy measurements on membranes: Simulations and experiments. Chemical Physics, 288, 171–186.

    Article  CAS  Google Scholar 

  61. Stauffer, T. P., & Meyer, T. (1997). Compartmentalized IgE receptor-mediated signal transduction in living cells. The Journal of Cell Biology, 139, 1447–1454.

    Article  PubMed  CAS  Google Scholar 

  62. Sheets, E. D., Holowka, D., & Baird, B. (1999). Critical role for cholesterol in Lyn-mediated tyrosine phosphorylation of Fc ε RI and their association with detergent-resistant membranes. The Journal of Cell Biology, 145, 877–887.

    Article  PubMed  CAS  Google Scholar 

  63. Pyenta, P. S., Holowka, D., & Baird, B. (2001). Cross-correlation analysis of inner-leaflet-anchored green fluorescent protein co-redistributed with IgE receptors and outer leaflet lipid raft components. Biophysical Journal, 80, 2120–2132.

    Article  PubMed  CAS  Google Scholar 

  64. Hebert, B., Hulme, S. E., & Wiseman, P. W. (2005). Membrane protein dynamics measured by two-photon ring correlation spectroscopy: Theory and application to living cells in multiphoton microscopy in the biomedical sciences V. In A. Periasamy & P. T. So (Eds.) Proceedings of S.P.I.E. (Vol. 5700, pp. 109–117).

  65. Petersen, N. O., Brown, C., Kaminski, A., Rocheleau, J., Srivastava, M., & Wiseman, P. W. (1998). Analysis of membrane protein cluster densities and sizes in situ by image correlation spectroscopy. Faraday Discuss (pp. 289–305); discussion 331–343.

  66. Ji, L., & Danuser, G. (2005). Tracking quasi-stationary flow of weak fluorescent signals by adaptive multi-frame correlation. Journal of Microscopy, 220, 150–167.

    Article  PubMed  CAS  Google Scholar 

  67. Adrian, R. J. (2005). Twenty years of particle image velocimetry. Experiments in Fluids, 39, 159–169.

    Article  Google Scholar 

  68. Gui, L., & Merzkirch, W. (2000). A comparative study of the MQD method and several correlation-based PIV evaluation algorithms. Experiments in Fluids, 28, 36–44.

    Article  Google Scholar 

  69. Brown, C. M., Hebert, B., Kolin, D. L., Zareno, J., Whitmore, L., Horwitz, A. F., & Wiseman, P. W. (2006). Probing the Integrin-Actin Linkage using High Resolution Protein Velocity Mapping. Journal of Cell Science, 119, 5204–5214.

    Article  PubMed  CAS  Google Scholar 

  70. Hebert, B. (2006). Spatio-temporal image correlation spectroscopy: Development and implementation in living cells Ph.D. thesis McGill University.

  71. Sinnecker, D., Voigt, P., Hellwig, N., & Schaefer, M. (2005). Reversible photobleaching of enhanced green fluorescent proteins. Biochemistry, 44, 7085–7094.

    Article  PubMed  CAS  Google Scholar 

  72. Dickson, R. M., Cubitt, A. B., Tsien, R. Y., & Moerner, W. E. (1997). On/Off blinking and switching behaviour of single molecules of green fluorescent protein. Nature, 388, 355–358.

    Article  PubMed  CAS  Google Scholar 

  73. Doose, S., Tsay, J. M., Pinaud, F., & Weiss, S. (2005). Comparison of photophysical and colloidal properties of biocompatible semiconductor nanocrystals using fluorescence correlation spectroscopy. Analytical Chemistry, 77, 2235–2242.

    Article  PubMed  CAS  Google Scholar 

  74. Bruchez, M., Moronne, M., Gin, P., Weiss, S., & Alivisatos, A. P. (1998). Semiconductor nanocrystals as fluorescent biological labels. Science, 281, 2013–2016.

    Article  PubMed  CAS  Google Scholar 

  75. Chan, W. C. W., & Nie, S. (1998). Quantum dot bioconjugates for ultrasensitive nonisotopic. Detection Science, 281, 2016–2018.

    CAS  Google Scholar 

  76. Yao, J., Larson, D. R., Vishwasrao, H. D., Zipfel, W. R., & Webb, W. W. (2005). Blinking and nonradiant dark fraction of water-soluble quantum dots in aqueous solution. Proceedings of the National Academy of Sciences of the United States of America, 102, 14284–14289.

    Article  PubMed  CAS  Google Scholar 

  77. Kuno, M., Fromm, D. P., Hamann, H. F., Gallagher, A., & Nesbitt, D. J. (2000). Nonexponential “blinking” kinetics of single CdSe quantum dots: A universal power law behavior. The Journal of Chemical Physics, 112, 3117–3120.

    Article  CAS  Google Scholar 

  78. Margolin, G., & Barkai, E. (2004). Aging correlation functions for blinking nanocrystals, and other on–off stochastic processes. The Journal of Chemical Physics, 121, 1566–1577.

    Article  PubMed  CAS  Google Scholar 

  79. Margolin, G., & Barkai, E. (2005). Nonergodicity of blinking nanocrystals and other Lévy-walk processes. Physical Review Letters, 94, 080601.

    Article  PubMed  CAS  Google Scholar 

  80. Kolin, D. L., Ronis, D., & Wiseman, P. W. (2006). k-Space image correlation spectroscopy: A method for accurate transport measurements independent of fluorophore photophysics. Biophysical Journal, 91, 3061–3075.

    Article  PubMed  CAS  Google Scholar 

  81. Durisic, N., Bachir, A. I., Kolin, D. L., Hebert, B., Lagerholm, B. C., Grutter, P., & Wiseman, P. W. (2007). Detection and correction of blinking bias in image correlation transport measurements of quantum dot tagged macromolecules. Biophysical Journal, 93, 1338–1346.

    Article  PubMed  CAS  Google Scholar 

  82. Berne, B. J., & Pecora, R. (2000). Dynamic light scattering. New York: Dover.

    Google Scholar 

  83. Hattori, M., Shimizu, H., & Yokoyama, H. (1996). Fluorescence correlation spectroscopy with traveling interference fringe excitation. The Review of Scientific Instruments, 67, 4064–4071.

    Article  CAS  Google Scholar 

  84. Digman, M. A., Sengupta, P., Wiseman, P. W., Brown, C. M., Horwitz, A. R., & Gratton, E. (2005). Fluctuation correlation spectroscopy with a laser-scanning microscope: Exploiting the hidden time structure. Biophysical Journal, 88, L33–L36.

    Article  PubMed  CAS  Google Scholar 

  85. Digman, M. A., Brown, C. M., Sengupta, P., Wiseman, P. W., Horwitz, A. R., & Gratton, E. (2005). Measuring fast dynamics in solutions and cells with a laser scanning microscope. Biophysical Journal, 89, 1317–1327.

    Article  PubMed  CAS  Google Scholar 

  86. Thompson, N. L. (1991). Fluorescence correlation spectroscopy in topics. In J. R. Lakowicz (Ed.), Fluorescence spectroscopy, volume 1: Techniques (pp. 337–378). New York: Plenum Press.

  87. Semrau, S., & Schmidt, T. (2007). Particle image correlation spectroscopy (PICS): Retrieving nanometer-scale correlations from high-density single-molecule position data. Biophysical Journal, 92, 613–621.

    Article  PubMed  CAS  Google Scholar 

  88. Brown, C. M. (2007). Fluorescence microscopy—avoiding the pitfalls. Journal of Cell Science, 120, 1703–1705.

    Article  PubMed  CAS  Google Scholar 

  89. Hibbs, A. R., MacDonald, G., & Garsha, K. (2006). Practical confocal microscopy. In J. B. Pawley (Ed.), Handbook of biological confocal microscopy (pp. 650–671). New York: Springer.

  90. Lauffenburger, D. A., & Horwitz, A. F. (1996). Cell migration: A physically integrated molecular process. Cell, 84, 359–369.

    Article  PubMed  CAS  Google Scholar 

  91. Webb, D. J., Parsons, J. T., & Horwitz, A. F. (2002). Adhesion assembly, disassembly and turnover in migrating cells—over and over and over again. Nature Cell Biology, 4, E97–E100.

    Article  PubMed  CAS  Google Scholar 

  92. Denk, W., Strickler, J. H., & Webb, W. W. (1990). Two-photon laser scanning fluorescence microscopy. Science, 248, 73–76.

    Article  PubMed  CAS  Google Scholar 

  93. Ridley, A. J., Schwartz, M. A., Burridge, K., Firtel, R. A., Ginsberg, M. H., Borisy, G., Parsons, J. T., & Horwitz, A. R. (2003). Cell migration: Integrating signals from front to back. Science, 302, 1704–1709.

    Article  PubMed  CAS  Google Scholar 

  94. Heldin, C.-H., Östman, A., & Rönnstrand, L. (1998). Signal transduction via platelet-derived growth factor receptors. Biochimica et Biophysica Acta, 1378, F79–F113.

    PubMed  CAS  Google Scholar 

  95. Yarden, Y., & Sliwkowski, M. X. (2001). Untangling the ErbB signalling network. Nature Reviews. Molecular Cell Biology, 2, 127–137.

    Article  PubMed  CAS  Google Scholar 

  96. Clayton, A. H. A., Walker, F., Orchard, S. G., Henderson, C., Fuchs, D., Rothacker, J., Nice, E. C., & Burgess, A. W. (2005). Ligand-induced dimer-tetramer transition during the activation of the cell surface epidermal growth factor receptor—A multidimensional microscopy analysis. The Journal of Biological Chemistry, 280, 30392–30399.

    Article  PubMed  CAS  Google Scholar 

  97. Saffarian, S., Li, Y., Elson, E. L., & Pike, L. J. (2007). Oligomerization of the EGF receptor investigated by live cell fluorescence intensity distribution analysis. Biophysical Journal, 93, 1021–1031.

    Article  PubMed  CAS  Google Scholar 

  98. Clayton, A. H. A., Tavarnesi, M. L., & Johns, T. G. (2007). Unligated epidermal growth factor receptor forms higher order oligomers within microclusters on A431 cells that are sensitive to tyrosine kinase inhibitor binding. Biochemistry, 46, 4589–4597.

    Article  PubMed  CAS  Google Scholar 

  99. Haag, R., & Kratz, F. (2006). Polymer therapeutics: Concepts and applications. Angewandte Chemie (International edition in English), 45, 1198–1215.

    Article  CAS  Google Scholar 

  100. Traub, L. M. (2003). Sorting it out: AP-2 and alternate clathrin adaptors in endocytic cargo selection. The Journal of Cell Biology, 163, 203–208.

    Article  PubMed  CAS  Google Scholar 

  101. Brown, C. M., & Petersen, N. O. (1998). An image correlation analysis of the distribution of clathrin associated adaptor protein (AP-2) at the plasma membrane. Journal of Cell Science, 111(Pt 2), 271–281.

    PubMed  CAS  Google Scholar 

  102. Brown, C. M., & Petersen, N. O. (1999). Free clathrin triskelions are required for the stability of clathrin-associated adaptor protein (AP-2) coated pit nucleation sites. Biochem. Cell Biol., 77, 439–448.

    Article  PubMed  CAS  Google Scholar 

  103. Brown, C. M., Roth, M. G., Henis, Y. I., & Petersen, N. O. (1999). An internalization-competent influenza hemagglutinin mutant causes the redistribution of AP-2 to existing coated pits and is colocalized with AP-2 in clathrin free clusters. Biochemistry, 38, 15166–15173.

    Article  PubMed  CAS  Google Scholar 

  104. Boyd, N. D., Chan, B. M. C., & Petersen, N. O. (2002). Adaptor protein-2 exhibits α1β1 or α6β1 integrin-dependent redistribution in rhabdomyosarcoma cells. Biochemistry, 41, 7232–7240.

    Article  PubMed  CAS  Google Scholar 

  105. Fire, E., Brown, C. M., Roth, M. G., Henis, Y. I., & Petersen, N. O. (1997). Partitioning of proteins into plasma membrane microdomains. Clustering of mutant influenza virus hemagglutinins into coated pits depends on the strength of the internalization signal. The Journal of Biological Chemistry, 272, 29538–29545.

    Article  PubMed  CAS  Google Scholar 

  106. Rasmusson, B. J., Flanagan, T. D., Turco, S. J., Epand, R. M., & Petersen, N. O. (1998). Fusion of Sendai virus and individual host cells and inhibition of fusion by lipophosphoglycan measured with image correlation spectroscopy. Biochimica et Biophysica Acta, 1404, 338–352.

    PubMed  CAS  Google Scholar 

  107. Rocheleau, J. V., & Petersen, N. O. (2000). Sendai virus binds to a dispersed population of NBD-GD1a. Bioscience Reports, 20, 139–155.

    Article  PubMed  CAS  Google Scholar 

  108. Rocheleau, J. V., & Petersen, N. O. (2001). The Sendai virus membrane fusion mechanism studied using image correlation spectroscopy. European Journal of Biochemistry, 268, 2924–2930.

    Article  PubMed  CAS  Google Scholar 

  109. Chen, D., Zhao, M., & Mundy, G. R. (2004). Bone morphogenetic proteins. Growth Factors, 22, 233–241.

    Article  PubMed  CAS  Google Scholar 

  110. Anderson, R. G. W. (1998). The caveolae membrane system. Annual Review of Biochemistry, 67, 199–225.

    Article  PubMed  CAS  Google Scholar 

  111. Nohe, A., Keating, E., Loh, C., Underhill, M. T., & Petersen, N. O. (2004a). Caveolin-1 isoform reorganization studied by image correlation spectroscopy. Faraday Discuss, 126, 185–195.

    Article  PubMed  CAS  Google Scholar 

  112. Nohe, A., Keating, E., Underhill, M. T., Knaus, P., & Petersen, N. O. (2004b). Dynamics and interaction of caveolin-1 isoforms with BMP-receptors. Journal of Cell Science, 118, 643–650.

    Article  CAS  Google Scholar 

  113. Nohe, A., Keating, E., Underhill, T. M., Knaus, P., & Petersen, N. O. (2003). Effect of the distribution and clustering of the type I A BMP receptor (ALK3) with the type II BMP receptor on the activation of signalling pathways. Journal of Cell Science, 116, 3277–3284.

    Article  PubMed  CAS  Google Scholar 

  114. Moser, M. B., Trommald, M., & Andersen, P. (1994). An increase in dendritic spine density on hippocampal CA1 pyramidal cells following spatial learning in adult rats suggests the formation of new synapses. Proceedings of the National Academy of Sciences of the United States of America, 91, 12673–12675.

    Article  PubMed  CAS  Google Scholar 

  115. Bates, I. R., Hebert, B., Luo, Y., Liao, J., Bashir, A. I., Kolin, D. L., Wiseman, P. W., & Hanrahan, J. W. (2006). Membrane lateral diffusion and capture of CFTR within transient confinement zones. Biophysical Journal, 91, 1046–1058.

    Article  PubMed  CAS  Google Scholar 

  116. Wang, M. D., & Axelrod, D. (1994). Microclustering patterns of acetylcholine receptors on myotubes studied by spatial fluorescence autocorrelation. Bioimaging, 2, 22–35.

    Article  CAS  Google Scholar 

  117. Hwang, J., Gheber, L. A., Margolis, L., & Edidin, M. (1998). Domains in cell plasma membranes investigated by near-field scanning optical microscopy. Biophysical Journal, 74, 2184–2190.

    Article  PubMed  CAS  Google Scholar 

  118. Tokumasu, F., Hwang, J., & Dvorak, J. A. (2004). Heterogeneous molecular distribution in supported multicomponent lipid bilayers. Langmuir, 20, 614–618.

    Article  PubMed  CAS  Google Scholar 

  119. Bachir, A. I., Durisic, N., Hebert, B., Grütter, P. H., & Wiseman, P. W. (2006). Image correlation spectroscopy studies of blinking dynamics in semiconductor quantum dots. Journal of Applied Physics, 99, 064503.

    Article  CAS  Google Scholar 

Download references

Acknowledgments

We thank the reviewers for their careful reading of the manuscript, and excellent suggestions. Additionally, DLK thanks the Natural Sciences and Engineering Research Council (NSERC) of Canada for a Canada Graduate Scholarship and PWW acknowledges NSERC and the Canadian Institutes of Health Research.

Author information

Authors and Affiliations

  1. Department of Chemistry, McGill University, Montreal, QC, Canada, H3A 2K6

    David L. Kolin & Paul W. Wiseman

  2. Department of Physics, McGill University, Montreal, QC, Canada, H3A 2T8

    Paul W. Wiseman

Authors
  1. David L. Kolin
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Paul W. Wiseman
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Paul W. Wiseman.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kolin, D.L., Wiseman, P.W. Advances in Image Correlation Spectroscopy: Measuring Number Densities, Aggregation States, and Dynamics of Fluorescently labeled Macromolecules in Cells. Cell Biochem Biophys 49, 141–164 (2007). https://doi.org/10.1007/s12013-007-9000-5

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12013-007-9000-5

Keywords

Search

Navigation