DOI QR코드

DOI QR Code

Statistical Analysis of Fluorescence Correlation Spectroscopy of Ultra Low Concentration Molecules with a Confocal Microscope

  • Received : 2008.06.23
  • Accepted : 2008.07.04
  • Published : 2008.09.25

Abstract

In this study, we simulated a statistical model of FCS(fluorescence correlation spectroscopy) based on a Poisson process to understand and explain observations of the experiment performed on molecules of ultra-low concentration by the home-built laser-scanning confocal microscope. The statistical model confirmed that the relative mean square amplitude of fluctuations is shown to be inversely proportional to the average number of molecules, even in the ultra-low concentration, if some conditions are satisfied. Signal-to-noise ratio and the variability of dwelling time under the confocal volume were found to be effective conditions for the experiment.

Keywords

References

  1. J. R. Lakowicz, Principles of Fluorescence Spectroscopy (Springer, New York, 2006), Chap. 24, pp. 798-806
  2. C. Gell and D. Brockwell, Handbook of Single Molecule Fluorescence Spectroscopy (Oxford, University Press, 2006), Chap. 2, pp. 25-26
  3. W. Becker, Advanced Time-correlated Single Photon Counting Techniques (Springer, New York, 2005), Chap. 5, pp. 178-180
  4. K. Bacia and P. Schwille, "Practical guidelines for dual -color fluorescence cross-correlation spectroscopy," Nature Protocols , vol. 2, no. 11, pp. 2842-2856, 2007 https://doi.org/10.1038/nprot.2007.410
  5. F. Fuji and M. Kinjo, "Detection of antigen protein by using fluorescence cross-correlation spectroscopy and Quantum-Dot-labeled antibodies," ChemBioChem. 8, pp. 2199-2203, 2007 https://doi.org/10.1002/cbic.200700399
  6. J. B. Pawley, Handbook of Biological Confocal Microscopy (Springer, New York, 2006), Chap. 7, pp. 145-157
  7. M. Wahl and I. Gregor, "Fast calculation of fluorescence correlation data with asynchronous time-correlated single-photon counting," Opt. Exp., vol. 11, issue 26, pp. 3583-3591, 2003 https://doi.org/10.1364/OE.11.003583