OSA's Digital Library

Optics Express

Optics Express

  • Editor: C. Martijn de Sterke
  • Vol. 20, Iss. 3 — Jan. 30, 2012
  • pp: 3043–3056

Single-laser shot fluorescence lifetime imaging on the nanosecond timescale using a Dual Image and Modeling Evaluation algorithm

Andreas Ehn, Olof Johansson, Andreas Arvidsson, Marcus Aldén, and Joakim Bood  »View Author Affiliations

Optics Express, Vol. 20, Issue 3, pp. 3043-3056 (2012)

View Full Text Article

Enhanced HTML    Acrobat PDF (1865 KB)

Browse Journals / Lookup Meetings

Browse by Journal and Year


Lookup Conference Papers

Close Browse Journals / Lookup Meetings

Article Tools



A novel technique, designated dual imaging and modeling evaluation (DIME), for evaluating single-laser shot fluorescence lifetimes is presented. The technique is experimentally verified in a generic gas mixing experiment to provide a clear demonstration of the rapidness and sensitivity of the detector scheme. Single-laser shot fluorescence lifetimes of roughly 800 ps with a standard deviation of ~120 ps were determined. These results were compared to streak camera measurements. Furthermore, a general fluorescence lifetime determination algorithm is proposed. The evaluation algorithm has an analytic, linear relationship between the fluorescence lifetime and detector signal ratio. In combination with the DIME detector scheme, it is a faster, more accurate and more sensitive approach for rapid fluorescence lifetime imaging than previously proposed techniques. Monte Carlo simulations were conducted to analyze the sensitivity of the detector scheme as well as to compare the proposed evaluation algorithm to previously presented rapid lifetime determination algorithms.

© 2012 OSA

OCIS Codes
(110.0110) Imaging systems : Imaging systems
(110.2970) Imaging systems : Image detection systems
(170.0170) Medical optics and biotechnology : Medical optics and biotechnology
(170.3650) Medical optics and biotechnology : Lifetime-based sensing
(110.4155) Imaging systems : Multiframe image processing
(110.3010) Imaging systems : Image reconstruction techniques

ToC Category:
Imaging Systems

Original Manuscript: August 30, 2011
Revised Manuscript: September 28, 2011
Manuscript Accepted: October 26, 2011
Published: January 25, 2012

Virtual Issues
Vol. 7, Iss. 3 Virtual Journal for Biomedical Optics

Andreas Ehn, Olof Johansson, Andreas Arvidsson, Marcus Aldén, and Joakim Bood, "Single-laser shot fluorescence lifetime imaging on the nanosecond timescale using a Dual Image and Modeling Evaluation algorithm," Opt. Express 20, 3043-3056 (2012)

Sort:  Author  |  Year  |  Journal  |  Reset  


  1. E. B. van Munster and T. W. Gadella, “Fluorescence lifetime imaging microscopy (FLIM),” Adv. Biochem. Eng. Biotechnol.95, 143–175 (2005). [PubMed]
  2. J. Lakowicz, Principles of Fluorescence Spectroscopy, 3rd ed. (Springer, 2006).
  3. A. D. Scully, R. B. Ostler, D. Phillips, P. O’Neill, K. M. S. Townsend, A. W. Parker, and A. J. MacRobert, “Application of fluorescence lifetime imaging microscopy to the investigation of intracellular PDT mechanisms,” Bioimaging5(1), 9–18 (1997). [CrossRef]
  4. R. Pepperkok, A. Squire, S. Geley, and P. I. H. Bastiaens, “Simultaneous detection of multiple green fluorescent proteins in live cells by fluorescence lifetime imaging microscopy,” Curr. Biol.9(5), 269–274 (1999). [CrossRef] [PubMed]
  5. P. J. Verveer, F. S. Wouters, A. R. Reynolds, and P. I. H. Bastiaens, “Quantitative imaging of lateral ErbB1 receptor signal propagation in the plasma membrane,” Science290(5496), 1567–1570 (2000). [CrossRef] [PubMed]
  6. H. E. Grecco, P. Roda-Navarro, A. Girod, J. Hou, T. Frahm, D. C. Truxius, R. Pepperkok, A. Squire, and P. I. H. Bastiaens, “In situ analysis of tyrosine phosphorylation networks by FLIM on cell arrays,” Nat. Methods7(6), 467–472 (2010). [CrossRef] [PubMed]
  7. C.-W. Chang, D. Sud, and M.-A. Mycek, “Fluorescence lifetime imaging microscopy,” Methods Cell Biol.81, 495–524 (2007). [CrossRef] [PubMed]
  8. T. Robinson, P. Valluri, H. B. Manning, D. M. Owen, I. Munro, C. B. Talbot, C. Dunsby, J. F. Eccleston, G. S. Baldwin, M. A. A. Neil, A. J. de Mello, and P. M. W. French, “Three-dimensional molecular mapping in a microfluidic mixing device using fluorescence lifetime imaging,” Opt. Lett.33(16), 1887–1889 (2008). [CrossRef] [PubMed]
  9. T. Ni and L. A. Melton, “Fuel equivalence ratio imaging for methane jets,” Appl. Spectrosc.47(6), 773–781 (1993). [CrossRef]
  10. T. Ni and L. A. Melton, “Two-dimensional gas-phase temperature measurements using fluorescence lifetime imaging,” Appl. Spectrosc. 50(9), 1112–1116 (1996). [CrossRef]
  11. A. Ehn, O. Johansson, J. Bood, A. Arvidsson, B. Li, and M. Aldén, “Fluorescence lifetime imaging in a flame,” Proc. Combust. Inst.33(1), 807–813 (2011). [CrossRef]
  12. W. Koban, J. D. Koch, R. K. Hanson, and C. Schulz, “Toluene LIF at elevated temperatures: implications for fuel–air ratio measurements,” Appl. Phys. B80(2), 147–150 (2005). [CrossRef]
  13. C. J. de Grauw and H. C. Gerritsen, “Multiple time-gate module for fluorescence lifetime imaging,” Appl. Spectrosc.55(6), 670–678 (2001). [CrossRef]
  14. D. V. O’Conner and D. Phillips, Time-Correlated Single Photon Counting (Academic, 1984).
  15. K. Dowling, S. C. W. Hyde, J. C. Dainty, P. M. W. French, and J. D. Hares, “2-D fluorescence lifetime imaging using a time-gated image intensifier,” Opt. Commun.135(1-3), 27–31 (1997). [CrossRef]
  16. X. F. Wang, T. Uchida, and S. Minami, “A fluorescence lifetime distribution measurement system based on phase-resolved detection using an image dissector tube,” Appl. Spectrosc.43(5), 840–845 (1989). [CrossRef]
  17. P. C. Schneider and R. M. Clegg, “Rapid acquisition, analysis, and display of fluorescence lifetime-resolved images for real-time application,” Rev. Sci. Instrum.68(11), 4107–4119 (1997). [CrossRef]
  18. R. J. Woods, S. Scypinski, and L. J. Cline Love “Transient digitizer for the determination of microsecond luminescence lifetimes,” Anal. Chem.56(8), 1395–1400 (1984). [CrossRef] [PubMed]
  19. D. S. Elson, I. Munro, J. Requejo-Isidro, J. McGinty, C. Dunsby, N. Galletly, G. W. Stamp, M. A. A. Neil, M. J. Lever, P. A. Kellett, A. Dymoke-Bradshaw, J. Hares, and P. M. W. French, “Real-time time-domain fluorescence lifetime imaging including single-shot acquisition with a segmented optical image intesifier,” New J. Phys.6, 180 (2004). [CrossRef]
  20. P. I. H. Bastiaens and A. Squire, “Fluorescence lifetime imaging microscopy: spatial resolution of biochemical processes in the cell,” Trends Cell Biol.9(2), 48–52 (1999). [CrossRef] [PubMed]
  21. G. Bunt and F. S. Wouters, “Visualization of molecular activities inside living cells with fluorescent labels,” Int. Rev. Cytol.237, 205–277 (2004). [CrossRef] [PubMed]
  22. A. Esposito, C. P. Dohm, M. Bähr, and F. S. Wouters, “Unsupervised fluorescence lifetime imaging microscopy for high content and high throughput screening,” Mol. Cell. Proteomics6(8), 1446–1454 (2007). [CrossRef] [PubMed]
  23. S. Kirkpatrick, C. D. Gelatt, and M. P. Vecchi, “Optimization by simulated annealing,” Science220(4598), 671–680 (1983). [CrossRef] [PubMed]
  24. T. B. Settersten and M. A. Linne, “Modeling pulsed excitation for gas-phase laser diagnostics,” J. Opt. Soc. Am. B19(5), 954–964 (2002). [CrossRef]
  25. R. A. Alberty and R. J. Silbey, Physical Chemistry, 2nd ed. (Wiley, New York, 1997), Chap 19.7.
  26. A. Elder, S. Schlachter, and C. F. Kaminski, “Theoretical investigation of the photon efficiency in frequency-domain fluorescence lifetime imaging microscopy,” J. Opt. Soc. Am. A25(2), 452–462 (2008). [CrossRef] [PubMed]
  27. A. Draaijer, R. Sanders, and H. C. Gerritsen, “Fluorescence lifetime imaging, a new tool in confocal microscopy,” in Handbook of Biological Confocal Microscopy, J. B. Pawley, ed. (Plenum, New York, 1995), pp. 491–505.
  28. J. McGinty, J. Requejo-Isidro, I. Munro, C. B. Talbot, P. A. Kellett, J. D. Hares, C. Dunsby, M A A. Neil, and P. M. W. French, “Signal-to-noise characterization of time-gated intensifiers used for wide-field time-domain FLIM,” J. Phys. D Appl. Phys.42(13), 135103 (2009). [CrossRef]
  29. A. V. Agronskaia, L. Tertoolen, and H. C. Gerritsen, “High frame rate fluorescence lifetime imaging,” J. Phys. D Appl. Phys.36(14), 1655–1662 (2003). [CrossRef]
  30. S. P. Chan, Z. J. Fuller, J. N. Demas, and B. A. DeGraff, “Optimized gating scheme for rapid lifetime determinations of single-exponential luminescence lifetimes,” Anal. Chem.73(18), 4486–4490 (2001). [CrossRef] [PubMed]

Cited By

Alert me when this paper is cited

OSA is able to provide readers links to articles that cite this paper by participating in CrossRef's Cited-By Linking service. CrossRef includes content from more than 3000 publishers and societies. In addition to listing OSA journal articles that cite this paper, citing articles from other participating publishers will also be listed.

« Previous Article  |  Next Article »

OSA is a member of CrossRef.

CrossCheck Deposited