OSA's Digital Library

Biomedical Optics Express

Biomedical Optics Express

  • Editor: Joseph A. Izatt
  • Vol. 2, Iss. 8 — Aug. 1, 2011
  • pp: 2255–2264

Cell assay using a two-photon-excited europium chelate

Xudong Xiao, Jeanne P. Haushalter, Kenneth T. Kotz, and Gregory W. Faris  »View Author Affiliations


Biomedical Optics Express, Vol. 2, Issue 8, pp. 2255-2264 (2011)
http://dx.doi.org/10.1364/BOE.2.002255


View Full Text Article

Enhanced HTML    Acrobat PDF (1186 KB)





Browse Journals / Lookup Meetings

Browse by Journal and Year


   


Lookup Conference Papers

Close Browse Journals / Lookup Meetings

Article Tools

Share
Citations

Abstract

We report application of two-photon excitation of europium chelates to immunolabeling of epidermal growth factor receptor (EGFR) cell surface proteins on A431 cancer cells. The europium chelates are excited with two photons of infrared light and emit in the visible. Europium chelates are conjugated to antibodies for EGFR. A431 (human epidermoid carcinoma) cells are labeled with this conjugate and imaged using a multiphoton microscope. To minimize signal loss due to the relatively long-lived Eu3+ emission, the multiphoton microscope is used with scanning laser two-photon excitation and non-scanning detection with a CCD. The chelate labels show very little photobleaching (less than 1% during continuous illumination in the microscope for 20 minutes) and low levels of autofluorescence (less than 1% of the signal from labeled cells). The detection limit of the europium label in the cell assay is better than 100 zeptomoles.

© 2011 OSA

OCIS Codes
(170.1530) Medical optics and biotechnology : Cell analysis
(170.2520) Medical optics and biotechnology : Fluorescence microscopy
(170.5810) Medical optics and biotechnology : Scanning microscopy
(190.7220) Nonlinear optics : Upconversion
(180.4315) Microscopy : Nonlinear microscopy

ToC Category:
Microscopy

History
Original Manuscript: May 2, 2011
Revised Manuscript: June 13, 2011
Manuscript Accepted: July 11, 2011
Published: July 13, 2011

Citation
Xudong Xiao, Jeanne P. Haushalter, Kenneth T. Kotz, and Gregory W. Faris, "Cell assay using a two-photon-excited europium chelate," Biomed. Opt. Express 2, 2255-2264 (2011)
http://www.opticsinfobase.org/boe/abstract.cfm?URI=boe-2-8-2255


Sort:  Author  |  Year  |  Journal  |  Reset  

References

  1. I. Hemmilä, S. Dakubu, V. M. Mukkala, H. Siitari, and T. Lövgren, “Europium as a label in time-resolved immunofluorometric assays,” Anal. Biochem. 137(2), 335–343 (1984). [CrossRef] [PubMed]
  2. R. C. Leif, R. A. Thomas, T. A. Yopp, B. D. Watson, V. R. Guarino, D. H. Hindman, N. Lefkove, and L. M. Vallarino, “Development of instrumentation and fluorochromes for automated multiparameter analysis of cells,” Clin. Chem. 23(8), 1492–1498 (1977). [PubMed]
  3. D. A. Zarling, M. Rossi, N. A. Peppers, J. Kane, G. W. Faris, M. J. Dyer, S. Y. Ng, and L. V. Schneider, “Up-converting reporters for biological and other assays using laser excitation techniques,” U.S. Patent 5,891,656 (6 April 1999).
  4. X. Xiao, J. P. Haushalter, and G. W. Faris, “Upconversion from aqueous phase lanthanide chelates,” Opt. Lett. 30(13), 1674–1676 (2005). [CrossRef] [PubMed]
  5. G. Piszczek, B. P. Maliwal, I. Gryczynski, J. Dattelbaum, and J. R. Lakowicz, “Multiphoton ligand-enhanced excitation of lanthanides,” J. Fluoresc. 11(2), 101–107 (2001). [CrossRef]
  6. X. Xiao, J. P. Haushalter, M. Weiss, and G. W. Faris, “Optical response measurement of upconverting luminescent reporters,” Proc. SPIE 5329, 229–238 (2004). [CrossRef]
  7. R. C. Leif, M. C. Becker, J. Alfred, J. Bromm, N. Chen, A. E. Cowan, L. M. Vallarino, S. Yang, and R. M. Zucker, “Lanthanide-enhanced luminescence (LEL) with one and two photon excitation of quantum dyes lanthanide(III)-macrocycles,” Proc. SPIE 5322, 187–199 (2004). [CrossRef]
  8. A. Picot, A. D’Aléo, P. L. Baldeck, A. Grichine, A. Duperray, C. Andraud, and O. Maury, “Long-lived two-photon excited luminescence of water-soluble europium complex: applications in biological imaging using two-photon scanning microscopy,” J. Am. Chem. Soc. 130(5), 1532–1533 (2008). [CrossRef] [PubMed]
  9. G. L. Law, K. L. Wong, C. W. Man, W. T. Wong, S. W. Tsao, M. H. Lam, and P. K. Lam, “Emissive terbium probe for multiphoton in vitro cell imaging,” J. Am. Chem. Soc. 130(12), 3714–3715 (2008). [CrossRef] [PubMed]
  10. G. Shao, R. Han, Y. Ma, M. Tang, F. Xue, Y. Sha, and Y. Wang, “Bionanoprobes with excellent two-photon-sensitized Eu3+ luminescence properties for live cell imaging,” Chemistry 16(29), 8647–8651 (2010). [CrossRef] [PubMed]
  11. C. Andraud and O. Maury, “Lanthanide complexes for nonlinear optics: from fundamental aspects to applications,” Eur. J. Inorg. Chem. 2009(29-30), 4357–4371 (2009). [CrossRef]
  12. L. O. Pålsson, R. Pal, B. S. Murray, D. Parker, and A. Beeby, “Two-photon absorption and photoluminescence of europium based emissive probes for bioactive systems,” Dalton Trans. (48): 5726–5734 (2007). [CrossRef] [PubMed]
  13. M. Bruchez, M. Moronne, P. Gin, S. Weiss, and A. P. Alivisatos, “Semiconductor nanocrystals as fluorescent biological labels,” Science 281(5385), 2013–2016 (1998). [CrossRef] [PubMed]
  14. W. C. W. Chan and S. Nie, “Quantum dot bioconjugates for ultrasensitive nonisotopic detection,” Science 281(5385), 2016–2018 (1998). [CrossRef] [PubMed]
  15. G. W. Faris, W. H. Wright, S. Pati, L. V. Schneider, and D. A. Zarling, “Upconverting reporters for biomedical diagnostics: Applications in antibody and DNA detection,” in Biomedical Optical Spectroscopy and Diagnostics, Vol. 3 of OSA Trends in Optics and Photonics, E. Sevick-Muraca and D. Benaron, eds. (Optical Society of America, Washington, DC, 1996), pp. 225–228.
  16. J. Hampl, M. P. Hall, N. A. Mufti, Y.-M. M. Yao, D. B. MacQueen, W. H. Wright, and D. E. Cooper, “Upconverting phosphor reporters in immunochromatographic assays,” Anal. Biochem. 288(2), 176–187 (2001). [CrossRef] [PubMed]
  17. A. Beeby, S. Faulkner, D. Parker, and J. A. G. Williams, “Sensitised luminescence from phenanthridine appended lanthanide complexes: analysis of triplet mediated energy transfer processes in terbium, europium and neodymium complexes,” J. Chem. Soc., Perkin Trans. 2 (8): 1268–1273 (2001). [CrossRef]
  18. L. D. Carlos, C. de Mello Donega, R. Q. Albuquerque, S. Alves, J. F. S. Menezes, and O. L. Malta, “Highly luminescent europium(III) complexes with naphtoiltrifluoroacetone and dimethyl sulphoxide,” Mol. Phys. 101(7), 1037–1045 (2003). [CrossRef]
  19. J. Squier and M. Muller, “High resolution nonlinear microscopy: A review of sources and methods for achieving optimal imaging,” Rev. Sci. Instrum. 72(7), 2855–2867 (2001). [CrossRef]
  20. J. Bewersdorf, R. Pick, and S. W. Hell, “Multifocal multiphoton microscopy,” Opt. Lett. 23(9), 655–657 (1998). [CrossRef] [PubMed]
  21. J. Mertz, C. Xu, and W. W. Webb, “Single-molecule detection by two-photon-excited fluorescence,” Opt. Lett. 20(24), 2532 (1995). [CrossRef] [PubMed]
  22. J. N. Mansbridge, W. A. Ausserer, M. A. Knapp, and R. M. Sutherland, “Adaptation of EGF receptor signal transduction to three-dimensional culture conditions: changes in surface receptor expression and protein tyrosine phosphorylation,” J. Cell. Physiol. 161(2), 374–382 (1994). [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