Abstract
A computational procedure is described for the analysis of fluorescence decay data convolved with a lamp flash of finite width. The computer program calculates the ratio of the Laplace transforms of the decay and the lamp flash for different values of s to give the transforms of the impulse response for each value of s. These are set equal to the analytical Laplace transforms of the decay law involved. Solution of the nonlinear simultaneous equations yields the desired decay parameters. The method can be modified to analyze data that contains a component due to scattered light and can also provide essential information regarding transit time changes of the photomultiplier with changes in emission wavelength. The method was tested by the analysis of real and simulated data. The accuracy of the analysis depends on the degree of correlation among the parameters.
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Selected References
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- Brand L., Gohlke J. R. Nanosecond time-resolved fluorescence spectra of a protein-dye complex. J Biol Chem. 1971 Apr 10;246(7):2317–2319. [PubMed] [Google Scholar]
- Grinvald A., Haas E., Steinberg I. Z. Evaluation of the distribution of distances between energy donors and acceptors by fluorescence decay. Proc Natl Acad Sci U S A. 1972 Aug;69(8):2273–2277. doi: 10.1073/pnas.69.8.2273. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Grinvald A., Steinberg I. Z. On the analysis of fluorescence decay kinetics by the method of least-squares. Anal Biochem. 1974 Jun;59(2):583–598. doi: 10.1016/0003-2697(74)90312-1. [DOI] [PubMed] [Google Scholar]
- Isenberg I., Dyson R. D., Hanson R. Studies on the analysis of fluorescence decay data by the method of moments. Biophys J. 1973 Oct;13(10):1090–1115. doi: 10.1016/S0006-3495(73)86047-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Isenberg I., Dyson R. D. The analysis of fluorescence decay by a method of moments. Biophys J. 1969 Nov;9(11):1337–1350. doi: 10.1016/S0006-3495(69)86456-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tung M. S., Steiner R. F. The application of nanosecond fluorometry to an allosteric protein. Biochem Biophys Res Commun. 1974 Apr 8;57(3):876–886. doi: 10.1016/0006-291x(74)90628-7. [DOI] [PubMed] [Google Scholar]
- Yashinsky G. Y. Fluorescence decay of lysozyme and of iodine oxidized lysozyme. FEBS Lett. 1972 Oct 1;26(1):123–126. doi: 10.1016/0014-5793(72)80556-8. [DOI] [PubMed] [Google Scholar]