Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Milestones
  • Published:

Milestone 10

FCS and FRAP: illuminating cellular processes

Fluorescence correlation spectroscopy, FRAP

Nature Cell Biology volume 11, pages S13–S14 (2009)Cite this article

Between 1972 and now, this technique, termed fluorescence correlation spectroscopy (FCS), has been applied to measure the kinetics of chemical and biological reactions, diffusion and flow. The rates of diffusion and flow were measured by detecting the stochastic movement of fluorophores into and out of a focused laser beam. Detection techniques have now improved, making it possible to quantify the dynamics of single molecules (Rigler and Widengren, 1990). The widespread adoption of FCS was initially constrained by the requirement for extensive calibration measurements and specialized equipment, but FCS modules for confocal microscopes are now available.

Twenty-five years after the initial studies using FCS, the development of fluorescence cross-correlation spectroscopy (FCCS) permitted the measurement of intermolecular interactions. In 1997, Schwille et al. labelled oligonucleotides with two discrete fluorophores — red and green — and measured the kinetics of red–green complex formation to deduce average annealing time. This technique has since been used to measure protein–protein interactions, including dynamic co-localization, trafficking and caspase-mediated cleavage.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

References

PRIMARY REFERENCES

  1. Magde, D., Elson, E. & Webb, W. W. Thermodynamic fluctuations in a reacting system — measurement by fluorescence correlation spectroscopy. Phys. Rev. Lett. 29, 705–708 (1972)

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  3. Koppel, E., Axelrod, D., Schlessinger, J., Elson, E. L. & Webb, W. W. Dynamics of fluorescence marker concentration as a probe of mobility. Biophys. J. 16, 1315–1329 (1976)

    Article  CAS  Google Scholar 

FURTHER READING

  1. Peters, R., Peters, J., Tews, K. H. & Bähr, W. A microfluorimetric study of translational diffusion in erythrocyte membranes. Biochim. Biophys. Acta 367, 282–294 (1974)

    Article  CAS  Google Scholar 

  2. Rigler, R. & Widengren, J. Ultrasensitive detection of single molecules by fluorescence correlation spectroscopy. Bioscience 3, 180–183 (1990)

    Google Scholar 

  3. Schwille, P., Meyer-Almes, F.-J. & Rigler, R. Dual-color fluorescence cross-correlation spectroscopy for multicomponent diffusional analysis in solution. Biophys. J. 72, 1878–1886 (1997)

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Associate Editor, UCSD-Nature Signaling Gateway,

    Emily J. Chenette

Authors
  1. Emily J. Chenette
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Chenette, E. FCS and FRAP: illuminating cellular processes. Nat Cell Biol 11 (Suppl 1), S13–S14 (2009). https://doi.org/10.1038/ncb1945

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/ncb1945

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing