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.

© 2008 Optical Society of Korea

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 [CrossRef]
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 [CrossRef]
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

ChemBioChem

• 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 [CrossRef]

Nature Protocols

• K. Bacia and P. Schwille, "Practical guidelines for dual -color fluorescence cross-correlation spectroscopy," Nature Protocols , vol. 2, no. 11, pp. 2842-2856, 2007 [CrossRef]

Opt. Exp.

• 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

Other

• J. B. Pawley, Handbook of Biological Confocal Microscopy (Springer, New York, 2006), Chap. 7, pp. 145-157
• J. R. Lakowicz, Principles of Fluorescence Spectroscopy (Springer, New York, 2006), Chap. 24, pp. 798-806
• C. Gell and D. Brockwell, Handbook of Single Molecule Fluorescence Spectroscopy (Oxford, University Press, 2006), Chap. 2, pp. 25-26
• W. Becker, Advanced Time-correlated Single Photon Counting Techniques (Springer, New York, 2005), Chap. 5, pp. 178-180

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