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Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells
Published on: February 9, 2012
Maximum entropy method for frequency-domain fluorescence lifetime analysis. 2. Timing, mismatched intensity, and
1Department of Chemistry, P.M. Gross Chemical Laboratory, Duke University, Durham, North Carolina 27708-0346, USA.
Analytical Chemistry
|February 15, 1996
Summary
The maximum entropy method (MEM) offers superior fluorescence lifetime recovery compared to nonlinear least-squares (NLLS) analysis, especially when dealing with systematic errors in frequency-domain measurements.
Area of Science:
- Spectroscopy and Photochemistry
- Data Analysis and Computational Methods
Background:
- Frequency-domain fluorescence lifetime measurements are susceptible to systematic errors.
- Standard nonlinear least-squares (NLLS) analysis can be sensitive to these errors.
- The maximum entropy method (MEM) offers a robust alternative for data analysis.
Purpose of the Study:
- To investigate the impact of common systematic errors on fluorescence lifetime recovery using MEM.
- To compare the performance of MEM against NLLS analysis in the presence of systematic errors.
- To evaluate the utility of MEM for frequency-domain fluorescence lifetime measurements.
Main Methods:
- Simultaneous minimization of chi-squared and maximization of a statistical entropy function.
- Analysis of real and simulated frequency-domain fluorescence lifetime data.
- Comparison of MEM and NLLS recovery of lifetimes, fractional intensities, and peak shapes.
Main Results:
- MEM demonstrated significant improvements over NLLS in recovering lifetimes and fractional intensities from data with timing or mismatched intensity errors.
- Both methods showed similar effects from reference lifetime errors in the presence of random noise.
- Characteristic changes in recovered parameters were linked to the identification of the true reference lifetime.
Conclusions:
- MEM provides a robust and unbiased approach to fluorescence lifetime data analysis.
- The dynamic, self-modeling nature of MEM, guided by the entropy criterion, enhances accuracy.
- MEM is a valuable tool for frequency-domain fluorescence lifetime recovery and other applications.
