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Title: An experimental comparison of the maximum likelihood estimation and nonlinear least squares fluorescence lifetime analysis of single molecules
Authors: Maus, MichaĆ«l ×
Cotlet, Mircea
Hofkens, Johan
Gensch, T
De Schryver, Frans
Schaffer, J
Seidel, CAM #
Issue Date: Jan-2001
Series Title: Analytical Chemistry vol:73 issue:9 pages:2078-2086
Abstract: Two procedures based on the weighted least-squares (LS) and the maximum likelihood estimation (MLE) method to confidently analyze single-molecule (SM! fluorescence decays with a total number (N) of 2500-60 000 counts have been elucidated and experimentally compared by analyzing measured bulk and SM decays. The key observation of this comparison is that the LS systematically underestimates the fluorescence lifetimes by similar to5%, for the range of 1000-20 000 events, whereas the MLE method gives stable results over the whole intensity range, even at counts N less than 1000, where the by analysis delivers unreasonable values. This difference can be attributed to the different statistics approaches and results from improper weighting of the LS method. As expected from theory, the results of both methods became equivalent above a certain threshold of N detected photons per decay, which is here experimentally determined to be similar to 20 000. In contrast to the hulk lifetime distributions, the SM fluorescence lifetime distributions exhibit standard deviations that are sizably larger than the statistically-expected values. This comparison proves the strong influence of the inhomogenuous microenvironment on the photophysical behavior of single molecules embedded in a 10-30-nm thin polymer layer.
URI: 
ISSN: 0003-2700
Publication status: published
KU Leuven publication type: IT
Appears in Collections:Chemistry - miscellaneous
Molecular Imaging and Photonics
× corresponding author
# (joint) last author

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