Statistical analysis and optimization of frequency-domain fluorescence lifetime imaging microscopy using homodyne lock-in detection
JOSA A, Vol. 27, Issue 5, pp. 1145-1155 (2010)
http://dx.doi.org/10.1364/JOSAA.27.001145
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Abstract
Fluorescence lifetime imaging microscopy is used widely in biological research, but the accuracy and precision of lifetime measurements are limited. Photon noise is an inherent error source that cannot be eliminated. In this paper, we present a general approach to compute the probability density of the estimated lifetime for frequency-domain fluorescence lifetime imaging microscopy using homodyne lock-in detection. The analysis for commonly used excitation methods, including sinusoidal modulation, square-wave modulation, and a periodically pulsed light source, are given and compared to the results of Monte Carlo simulations. The optimum parameters of the excitation waveforms to minimize the variance of the estimated lifetimes are also derived and compared to previously published results.
© 2010 Optical Society of America
OCIS Codes
(000.5490) General : Probability theory, stochastic processes, and statistics
(030.4280) Coherence and statistical optics : Noise in imaging systems
(030.5290) Coherence and statistical optics : Photon statistics
(170.1790) Medical optics and biotechnology : Confocal microscopy
(170.2520) Medical optics and biotechnology : Fluorescence microscopy
(170.3650) Medical optics and biotechnology : Lifetime-based sensing
ToC Category:
Medical Optics and Biotechnology
History
Original Manuscript: November 11, 2009
Manuscript Accepted: February 25, 2010
Published: April 28, 2010
Virtual Issues
Vol. 5, Iss. 9 Virtual Journal for Biomedical Optics
Citation
Yuxiang Lin and Arthur F. Gmitro, "Statistical analysis and optimization of frequency-domain fluorescence lifetime imaging microscopy using homodyne lock-in detection," J. Opt. Soc. Am. A 27, 1145-1155 (2010)
http://www.opticsinfobase.org/josaa/abstract.cfm?URI=josaa-27-5-1145
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