OSA's Digital Library

Virtual Journal for Biomedical Optics

Virtual Journal for Biomedical Optics


  • Editor: Gregory W. Faris
  • Vol. 5, Iss. 5 — Mar. 17, 2010

Imaging live cell membranes via surface plasmon-enhanced fluorescence and phase microscopy

Ruei-Yu He, Chun-Yu Lin, Yuan-Deng Su, Kuo-Chih Chiu, Nan-Shan Chang, Hua-Lin Wu, and Shean-Jen Chen  »View Author Affiliations

Optics Express, Vol. 18, Issue 4, pp. 3649-3659 (2010)

View Full Text Article

Enhanced HTML    Acrobat PDF (1510 KB) Open Access

Browse Journals / Lookup Meetings

Browse by Journal and Year


Lookup Conference Papers

Close Browse Journals / Lookup Meetings

Article Tools



This paper demonstrates the first combination for wide-field surface plasmon (SP) phase microscopy and SP-enhanced fluorescence microscopy to image living cells’ contacts on the surface of a bio-substrate simultaneously. The phase microscopy with a phase-shift interferometry and common-path optical setup can provide high-sensitivity phase information in long-term stability. Simultaneously, the fluorescence microscopy with the enhancement of a local electromagnetic field can supply bright fluorescent images. The combined microscope imposes a high numerical aperture objective upon the excitation of surface plasmon through a silver film with a thickness of 30 nm. The developed SP microscope is successfully applied to the real-time bright observation of the transfected fluorescence of living cells localized near the cell membrane on the bio-substrate and the high-sensitivity phase image of the cell-substrate contacts at the same time.

© 2010 OSA

OCIS Codes
(170.1530) Medical optics and biotechnology : Cell analysis
(170.2520) Medical optics and biotechnology : Fluorescence microscopy
(240.6680) Optics at surfaces : Surface plasmons

ToC Category:
Medical Optics and Biotechnology

Original Manuscript: January 5, 2010
Revised Manuscript: January 31, 2010
Manuscript Accepted: February 3, 2010
Published: February 4, 2010

Virtual Issues
Vol. 5, Iss. 5 Virtual Journal for Biomedical Optics

Ruei-Yu He, Chun-Yu Lin, Yuan-Deng Su, Kuo-Chih Chiu, Nan-Shan Chang, Hua-Lin Wu, and Shean-Jen Chen, "Imaging live cell membranes via surface plasmon-enhanced fluorescence and phase microscopy," Opt. Express 18, 3649-3659 (2010)

Sort:  Author  |  Year  |  Journal  |  Reset  


  1. G. A. Truskey, J. S. Burmeister, E. Grapa, and W. M. Reichert, “Total internal reflection fluorescence microscopy (TIRFM). II. Topographical mapping of relative cell/substratum separation distances,” J. Cell Sci. 103(Pt 2), 491–499 (1992). [PubMed]
  2. S. E. Sund and D. Axelrod, “Actin dynamics at the living cell submembrane imaged by total internal reflection fluorescence photobleaching,” Biophys. J. 79(3), 1655–1669 (2000). [CrossRef] [PubMed]
  3. W. J. Betz, F. Mao, and C. B. Smith, “Imaging exocytosis and endocytosis,” Curr. Opin. Neurobiol. 6(3), 365–371 (1996). [CrossRef] [PubMed]
  4. D. Axelrod, “Total internal reflection fluorescence microscopy in cell biology,” Methods Enzymol. 361, 1–33 (2003). [CrossRef] [PubMed]
  5. K. F. Giebel, C. Bechinger, S. Herminghaus, M. Riedel, P. Leiderer, U. Weiland, and M. Bastmeyer, “Imaging of cell/substrate contacts of living cells with surface plasmon resonance microscopy,” Biophys. J. 76(1), 509–516 (1999). [CrossRef] [PubMed]
  6. H. Raether, Surface Plasmons on Smooth and Rough Surfaces and on Gratings (Springer, 1998).
  7. B. Huang, F. Yu, and R. N. Zare, “Surface plasmon resonance imaging using a high numerical aperture microscope objective,” Anal. Chem. 79(7), 2979–2983 (2007). [CrossRef] [PubMed]
  8. M. M. A. Jamil, M. C. T. Denyer, M. Youseffi, S. T. Britland, S. Liu, C. W. See, M. G. Somekh, and J. Zhang, “Imaging of the cell surface interface using objective coupled widefield surface plasmon microscopy,” J. Struct. Biol. 164(1), 75–80 (2008). [CrossRef] [PubMed]
  9. K. J. Moh, X. C. Yuan, J. Bu, S. W. Zhu, and B. Z. Gao, “Surface plasmon resonance imaging of cell-substrate contacts with radially polarized beams,” Opt. Express 16(25), 20734–20741 (2008). [CrossRef] [PubMed]
  10. Y. D. Su, S. J. Chen, and T. L. Yeh, “Common-path phase-shift interferometry surface plasmon resonance imaging system,” Opt. Lett. 30(12), 1488–1490 (2005). [CrossRef] [PubMed]
  11. K. H. Lee, Y. D. Su, S. J. Chen, F. G. Tseng, and G. B. Lee, “Microfluidic systems integrated with two-dimensional surface plasmon resonance phase imaging systems for microarray immunoassay,” Biosens. Bioelectron. 23(4), 466–472 (2007). [CrossRef] [PubMed]
  12. R. Y. He, G. L. Chang, H. L. Wu, C. H. Lin, K. C. Chiu, Y. D. Su, and S. J. Chen, “Enhanced live cell membrane imaging using surface plasmon-enhanced total internal reflection fluorescence microscopy,” Opt. Express 14(20), 9307–9316 (2006). [CrossRef] [PubMed]
  13. R.-Y. He, Y.-D. Su, K.-C. Cho, C.-Y. Lin, N.-S. Chang, C.-H. Chang, and S.-J. Chen, “Surface plasmon-enhanced two-photon fluorescence microscopy for live cell membrane imaging,” Opt. Express 17(8), 5987–5997 (2009). [CrossRef] [PubMed]
  14. A. W. Peterson, M. Halter, A. Tona, K. Bhadriraju, and A. L. Plant, “Surface plasmon resonance imaging of cells and surface-associated fibronectin,” BMC Cell Biol. 10(1), 16 (2009). [CrossRef] [PubMed]
  15. B. Rothenhäusler and W. Knoll, “Interferometric determination of the complex wave vector of plasmon surface polaritons,” J. Opt. Soc. Am. B 5(7), 1401–1405 (1988). [CrossRef]
  16. W. Knoll, “Optical characterization of organic thin films and interfaces with evanescent waves,” Mat. Res. Soc. Bulletin. 16, 29–39 (1991).
  17. C. D. Geddes and J. R. Lakowicz, “Metal-enhanced fluorescence,” J. Fluoresc. 12(2), 121–129 (2002). [CrossRef]
  18. J. R. Lakowicz, “Radiative decay engineering: biophysical and biomedical applications,” Anal. Biochem. 298(1), 1–24 (2001). [CrossRef] [PubMed]
  19. N. S. Chang, N. Pratt, J. Heath, L. Schultz, D. Sleve, G. B. Carey, and N. Zevotek, “Hyaluronidase induction of a WW domain-containing oxidoreductase that enhances tumor necrosis factor cytotoxicity,” J. Biol. Chem. 276(5), 3361–3370 (2001). [CrossRef]
  20. N. S. Chang, L. J. Hsu, Y. S. Lin, F. J. Lai, and H. M. Sheu, “WW domain-containing oxidoreductase: a candidate tumor suppressor,” Trends Mol. Med. 13(1), 12–22 (2007). [CrossRef]
  21. C. E. H. Berger, R. P. H. Kooyman, and J. Greve, “Resolution in surface plasmon microscopy,” Rev. Sci. Instrum. 65(9), 2829–2836 (1994). [CrossRef]
  22. P. Anger, P. Bharadwaj, and L. Novotny, “Enhancement and quenching of single-molecule fluorescence,” Phys. Rev. Lett. 96(11), 113002 (2006). [CrossRef] [PubMed]
  23. P. Hariharan, B. F. Oreb, and T. Eiju, “Digital phase-shifting interferometry: a simple error-compensating phase calculation algorithm,” Appl. Opt. 26(13), 2504–2505 (1987). [CrossRef] [PubMed]
  24. J.-J. Chyou, S.-J. Chen, and Y.-K. Chen, “Two-dimensional phase unwrapping with a multichannel least-mean-square algorithm,” Appl. Opt. 43(30), 5655–5661 (2004). [CrossRef] [PubMed]
  25. Q. Hong, L. J. Hsu, L. Schultz, N. Pratt, J. Mattison, and N. S. Chang, “Zfra affects TNF-mediated cell death by interacting with death domain protein TRADD and negatively regulates the activation of NF-kappaB, JNK1, p53 and WOX1 during stress response,” BMC Mol. Biol. 8(1), 50 (2007). [CrossRef] [PubMed]
  26. R. M. Fulbright and D. Axelrod, “Dynamics of nonspecific adsorption of insulin to erythrocyte membranes,” J. Fluoresc. 3(1), 1–16 (1993). [CrossRef]
  27. B. Rothenhäusler and W. Knoll, “Surface-plasmon microscopy,” Nature 332(6165), 615–617 (1988). [CrossRef]
  28. M. C. Boffa, B. Burke, and C. C. Haudenschild, “Preservation of thrombomodulin antigen on vascular and extravascular surfaces,” J. Histochem. Cytochem. 35(11), 1267–1276 (1987). [CrossRef] [PubMed]
  29. H. C. Huang, G. Y. Shi, S. J. Jiang, C. S. Shi, C. M. Wu, H. Y. Yang, and H. L. Wu, “Thrombomodulin-mediated cell adhesion: involvement of its lectin-like domain,” J. Biol. Chem. 278(47), 46750–46759 (2003). [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.


Fig. 1 Fig. 2 Fig. 3
Fig. 4 Fig. 5

« Previous Article  |  Next Article »

OSA is a member of CrossRef.

CrossCheck Deposited