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Optics Letters

Optics Letters


  • Vol. 28, Iss. 21 — Nov. 1, 2003
  • pp: 2073–2075

Combined Raman and continuous-wave-excited two-photon fluorescence cell imaging

Natallia Uzunbajakava and Cees Otto  »View Author Affiliations

Optics Letters, Vol. 28, Issue 21, pp. 2073-2075 (2003)

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We demonstrate a confocal optical microscope that combines cw two-photon-excited fluorescence microscopy with confocal Raman microscopy. With this microscope fast image acquisition with fluorescence imaging can be used to select areas of interest for subsequent chemical analysis with spontaneous Raman imaging. The distribution of the UV-absorbing fluorophore Hoechst 33342 in the apoptotic HeLa cells is measured in the combined cw two-photon-excited fluorescence and Raman microscopy modes. The 647-nm line of a Kr-ion laser is used to excite both the Raman scattering and the two-photon-excited fluorescence emission. The lateral and axial resolutions in the two imaging modes are compared by use of the Gaussian beam approximation and backprojection of the focal volume through the confocal pinhole.

© 2003 Optical Society of America

OCIS Codes
(170.5660) Medical optics and biotechnology : Raman spectroscopy
(180.1790) Microscopy : Confocal microscopy
(300.6410) Spectroscopy : Spectroscopy, multiphoton
(300.6450) Spectroscopy : Spectroscopy, Raman

Natallia Uzunbajakava and Cees Otto, "Combined Raman and continuous-wave-excited two-photon fluorescence cell imaging," Opt. Lett. 28, 2073-2075 (2003)

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  1. J.-X. Cheng, Y. Jia, G. Zheng, and X. Xie, Biophys. J. 83, 502 (2002).
  2. G. Puppels, H. Garritsen, J. Kummer, and J. Greve, Cytometry 14, 251 (1993).
  3. R. Ramanauskaite, G. Segers-Nolten, K. de Grauw, N. Sijtsema, L. van der Maas, J. Greve, and C. Otto, C. Figdor, Pure Appl. Chem. 69, 2131 (1997).
  4. S. Arikan, H. Sands, R. Rodway, and D. Batchelder, Anim. Reprod. Sci. 71, 249 (2002).
  5. N. Uzunbajakava, A. Lenferink, Y. Kraan, B. Willekens, G. Vrensen, J. Greve, and C. Otto, Biopolymers 72, 1 (2003).
  6. N. Uzunbajakava, A. Lenferink, Y. Kraan, E. Volokhina, G. Vrensen, J. Greve, and C. Otto, Biophys. J. 84, 3968 (2003).
  7. W. Denk, J. Strickler, and W. Webb, Science 248, 73 (1990).
  8. S. Hell, M. Booth, and S. Wilms, Opt. Lett. 23, 1238 (1998).
  9. M. Booth and S. Hell, J. Microsc. 190, 298 (1997).
  10. A. Kirsch, V. Subramaniam, G. Striker, C. Schnetter, D. Arndt-Jovin, and T. Jovin, Biophys. J. 75, 1513 (1998).
  11. Y. Liu, G. Sonek, M. W. Berns, K. Konig, and B. J. Tromberg, Opt. Lett. 20, 2246 (1995).
  12. J. Kerr, A. Wyllie, and A. Curie, Br. J. Cancer 26, 239 (1972).
  13. F. Zimmermann, B. Zimmermann, J. Panitz, and A. Wokaun, J. Raman Spectrosc. 26, 435 (1995).
  14. H. Deng, V. Bloomfield, J. Benevides, and G. Thomas, Biopolymers 50, 656 (1999).
  15. W. Peticolas, W. Kubasek, G. Thomas, and M. Tsuboi, in Biological Applications of Raman Spectroscopy, T. Spiro, ed. (Wiley, New York, 1987), Vol. 1, pp. 81–134.
  16. F. van der Heijden, Image Based Measurement Systems: Object Recognition and Parameter Estimation (Wiley, New York, 1994).
  17. Z. Darzynkiewicz, in Flow Cytometry and Sorting, M. Melamed, T. Lindmo, and M. Mendelsohn, eds. (Wiley, New York, 1990), pp. 315–340.
  18. C. de Grauw, N. Sijtsema, C. Otto, and J. Greve, J. Microsc. 188, 273 (1997).
  19. C. Xu W. Webb, in Topics in Fluorescence Spectroscopy, J. Lakowicz, ed. (Plenum, New York, 1997), Vol. 5, pp. 472–540.
  20. C. Sheppard and M. Gu, Optik 86, 104 (1990).
  21. S. Hell and E. Stelzer, Opt. Commun. 93, 277 (1992).

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