OSA's Digital Library

Applied Optics

Applied Optics


  • Editor: James C. Wyant
  • Vol. 45, Iss. 20 — Jul. 10, 2006
  • pp: 5020–5026

Subnanosecond-resolution phase-resolved fluorescence imaging technique for biomedical applications

U. S. Dinish, C. Y. Fu, Z. X. Chao, L. K. Seah, V. M. Murukeshan, and B. K. Ng  »View Author Affiliations

Applied Optics, Vol. 45, Issue 20, pp. 5020-5026 (2006)

View Full Text Article

Enhanced HTML    Acrobat PDF (862 KB)

Browse Journals / Lookup Meetings

Browse by Journal and Year


Lookup Conference Papers

Close Browse Journals / Lookup Meetings

Article Tools



Characterization of fluorescence emissions from cells often leads to conclusive results in the early detection of cellular abnormalities. Cellular abnormalities can be characterized by their difference in the fluorescence lifetime, which may be less than nanoseconds. A sensitive frequency domain technique, also called a phase-resolved fluorescence imaging technique, is proposed in which fluorescence emissions at the same wavelengths can more effectively be separated with subnanosecond resolution in their lifetime difference. The system configuration is optimized by incorporating even-step phase shifting in the homodyne-assisted signal-processing concept along with the phase-resolved fluorescence technique to eliminate the dc offsets of emission. Experiments are carried out with simulated samples composed of two fluorescence emissions of the same wavelength but with different lifetime values. Suppression of either of the fluorescence emissions by selective imaging of the other validates the superiority of the proposed technique. Hence, this technique can potentially be applied in the early detection of cellular abnormalities.

© 2006 Optical Society of America

OCIS Codes
(170.0110) Medical optics and biotechnology : Imaging systems
(300.2530) Spectroscopy : Fluorescence, laser-induced

Original Manuscript: July 18, 2005
Revised Manuscript: January 17, 2006
Manuscript Accepted: April 3, 2006

Virtual Issues
Vol. 1, Iss. 8 Virtual Journal for Biomedical Optics

U. S. Dinish, C. Y. Fu, Z. X. Chao, L. K. Seah, V. M. Murukeshan, and B. K. Ng, "Subnanosecond-resolution phase-resolved fluorescence imaging technique for biomedical applications," Appl. Opt. 45, 5020-5026 (2006)

Sort:  Author  |  Year  |  Journal  |  Reset  


  1. J. R. Lakowicz, Principles of Fluorescence Spectroscopy (Plenum, 1999).
  2. G. G. Guilbault, Fluorescence: Theory, Instrumentation and Practice (Edward Arnold, 1967).
  3. J. P. Houston, A. B. Thompson, M. Gurfinkel, and E. M. Sevick-Muraca, "Sensitivity and depth penetration of continuous wave versus frequency-domain photon migration near-infrared fluorescence contrast-enhanced imaging," Photochem. Photobiol. 77, 420-430 (2003). [CrossRef] [PubMed]
  4. A. Godavarty, M. J. Eppstein, C. Zhang, S. Theru, A. B. Thompson, M. Gurfinkel, and E. M. Sevick-Muraca, "Fluorescence-enhanced optical imaging in large tissue volumes using a gain-modulated ICCD camera," Phys. Med. Biol. 48, 1701-1720 (2003). [CrossRef] [PubMed]
  5. S. Lam, C. MacAulay, J. Leriche, N. Ikeda, and B. Palcic, "Fluorescence imaging of early lung cancer," in Optical Biopsy and Fluorescence Spectroscopy and Imaging,Proc. SPIE 2324, 2-8 (1995). [CrossRef]
  6. D. Erimberger, D. Zaak, H. Stepp, R. Knuchel, R. Baumgartner, P. Schneede, N. Schmeller, and A. Hofstetter, "Auto fluorescence imaging to optimise 5-ALA-induced fluorescence endoscopy of bladder carcinoma," Urology 58, 372-375 (2001). [CrossRef]
  7. J. Sipior, G. M. Carter, J. R. Lakowicz, and G. Rao, "Blue light-emitting diode demonstrated as an ultraviolet excitation source for nanosecond phase-modulation fluorescence lifetime measurements," Rev. Sci. Instrum. 68, 2666-2670 (1997). [CrossRef]
  8. T. Arakia and H. Misawa, "Light emitting diode-based nanosecond ultraviolet light source for fluorescence lifetime measurements," Rev. Sci. Instrum. 66, 5469-5472 (1995). [CrossRef]
  9. C. G. Morgan, Y. Hua, A. C. Mitchell, J. G. Murray, and A. D. Boardman, "A compact frequency domain fluorometer with a directly modulated deuterium light source," Rev. Sci. Instrum. 67, 41-47 (1996). [CrossRef]
  10. Z. Zhang, K. T. V. Grattan, and A. W. Palmer, "Phase-locked detection of fluorescence lifetime," Rev. Sci. Instrum. 64, 2531-2540 (1993). [CrossRef]
  11. V. Venkatesh and V. Srinivas, "A closed loop scheme for phase-sensitive fluorometry," Rev. Sci. Instrum. 66, 3750-3754 (1995). [CrossRef]
  12. R. V. Krishnan, H. Saitoh, H. Terada, V. E. Centonze, and B. Hermana, "Development of a multiphoton fluorescence lifetime imaging microscopy system using a streak camera," Rev. Sci. Instrum. 74, 2714-2721 (2003). [CrossRef]
  13. J. R. Lakowicz and K. W. Berndt, "Lifetime-selective fluorescence imaging using an rf phase-sensitive camera," Rev. Sci. Instrum. 62, 1727-1734 (1991). [CrossRef]
  14. U. S. Dinish, Z. X. Chao, A. Singh, L. K. Seah, and V. M. Murukeshan, "Formulation and implementation of phase-resolved optical technique for latent fingerprint imaging: theoretical and experimental analysis," Appl. Opt. 44, 297-304 (2005). [CrossRef] [PubMed]
  15. L. B. Mcgown and D. W. Millican, "Multifrequency phase-resolution for total luminescence spectroscopy," in Time-Resolved Laser Spectroscopy in Biochemistry,Proc. SPIE 909, 360-365 (1988).
  16. Z. X. Chao, U. S. Dinish, L. K. Seah, and V. M. Murukeshan, "Homodyne and heterodyne signal processing assisted phase resolved optical technique for latent fingerprint imaging: a theoretical study," J. Mod. Opt. 52, 119-129 (2005). [CrossRef]
  17. B. C. MacDonald, S. J. Lvin, and H. Patterson, "Correction of fluorescence inner filter effects and the partitioning of pyrene to dissolved organic carbon," Anal. Chim. Acta 338, 155-162 (1997). [CrossRef]
  18. A. Pradhan, B. B. Das, K. M. Yoo, R. R. Alfano, J. M. D. Cleary, R. M. D. Prudente, and E. M. D. Celmer, "Time-resolved fluorescence of benign and malignant breast tissues," Proc. SPIE 1599, 81-84 (1991). [CrossRef]

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.


Fig. 1 Fig. 2 Fig. 3
Fig. 4

« Previous Article  |  Next Article »

OSA is a member of CrossRef.

CrossCheck Deposited