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Journal of the Optical Society of America A

Journal of the Optical Society of America A

| OPTICS, IMAGE SCIENCE, AND VISION

  • Editor: Stephen A. Burns
  • Vol. 25, Iss. 2 — Feb. 1, 2008
  • pp: 452–462

Theoretical investigation of the photon efficiency in frequency-domain fluorescence lifetime imaging microscopy

Alan Elder, Simon Schlachter, and Clemens F. Kaminski  »View Author Affiliations


JOSA A, Vol. 25, Issue 2, pp. 452-462 (2008)
http://dx.doi.org/10.1364/JOSAA.25.000452


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Abstract

We investigate the photon efficiency of frequency-domain fluorescence lifetime imaging microscopy, using both theoretical and Monte Carlo methods. Our analysis differs from previous work in that it incorporates the data fitting process used in real experiments, allows for the arbitrary choice of excitation and gain waveforms, and calculates lifetimes as well as associated F-values from higher harmonics in the data. Using our analysis, we found different photon efficiencies to those previously reported and were able to propose optimal excitation and gain waveforms. Additionally, we suggest measurement protocols that lead to further improvement in photon efficiency. We compare our results to other techniques for lifetime imaging and consider the implications of our higher-harmonic analysis for multi-exponential lifetime determination.

© 2008 Optical Society of America

OCIS Codes
(000.5490) General : Probability theory, stochastic processes, and statistics
(170.2520) Medical optics and biotechnology : Fluorescence microscopy
(170.3650) Medical optics and biotechnology : Lifetime-based sensing
(180.2520) Microscopy : Fluorescence microscopy

ToC Category:
Fourier Optics and Signal Processing

History
Original Manuscript: July 6, 2007
Revised Manuscript: November 18, 2007
Manuscript Accepted: December 11, 2007
Published: January 29, 2008

Virtual Issues
Vol. 3, Iss. 3 Virtual Journal for Biomedical Optics

Citation
Alan Elder, Simon Schlachter, and Clemens F. Kaminski, "Theoretical investigation of the photon efficiency in frequency-domain fluorescence lifetime imaging microscopy," J. Opt. Soc. Am. A 25, 452-462 (2008)
http://www.opticsinfobase.org/josaa/abstract.cfm?URI=josaa-25-2-452

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