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Statistical accuracy in fluorescence correlation spectroscopy
Abstract
Fluorescence correlation spectroscopy is a promising technique for the study of chemical kinetics and diffusion processes in small, well-defined sample volumes. Laser-induced fluorescence is used as a specific highly sensitive probe of concentration, permitting the analysis of the characteristic time dependence of spontaneous concentration fluctuations. We present here a quantitative analysis of the statistical and systematic errors inherent in such an experiment. Theoretical expressions are derived for the signal-to-noise ratio for various operational definitions of the photocount correlation signal. It is shown that the critical parameter governing the statistical accuracy of the experiment is not the total number of photocounts per correlation time, but (when background fluorescence is negligible) the photocounts per correlation time, per fluorescent molecule, a parameter independent of the number of fluorescent molecules in the sample volume. Comparisons are made with parallel results for laser-light scattering experiments derived by other authors. Finally, we consider the effects of background fluorescence, photolysis, and other deviations from ideality.
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Cited by
Mobility measurement by analysis of fluorescence photobleaching recovery kinetics
Axelrod, D., Koppel, D. E., Schlessinger, J., Elson, E., & Webb, W. W. (1976) · Biophysical Journal · doi:10.1016/s0006-3495(76)85755-4 · Cited by 2,714
Self-citationZero-Mode Waveguides for Single-Molecule Analysis at High Concentrations
Levene, M. J., Korlach, J., Turner, S. W., Foquet, M., Craighead, H. G., & Webb, W. W. (2003) · Science · doi:10.1126/science.1079700 · Cited by 1,378
Valence and patterning of aromatic residues determine the phase behavior of prion-like domains
Martin, E. W., Holehouse, A. S., Peran, I., Farag, M., Incicco, J. J., Bremer, A., Grace, C. R., Soranno, A., Pappu, R. V., & Mittag, T. (2020) · Science · doi:10.1126/science.aaw8653 · Cited by 1,147
Probing Individual Molecules with Confocal Fluorescence Microscopy
Nie, S., Chiu, D. T., & Zare, R. N. (1994) · Science · doi:10.1126/science.7973650 · Cited by 549
Probing Lipid Mobility of Raft-exhibiting Model Membranes by Fluorescence Correlation Spectroscopy
Kahya, N., Scherfeld, D., Bacia, K., Poolman, B., & Schwille, P. (2003) · Journal of Biological Chemistry · doi:10.1074/jbc.m302969200 · Cited by 499
Metadata
- Authors
- Koppel, D. E.
- Venue
- Physical Review A
- Publisher
- American Physical Society (APS)
- Published
- 1974-12-01
- Volume
- 10
- Issue
- 6
- Pages
- 1938-1945
- ISSN
- 0556-2791 (ISSN-L)
- OpenAlex
- W2064757943
- Cited by
- 450
- References
- 0
- FWCI
- 3.39
- Type
- journal article
- Language
- en
- Open access
- closed
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