Abstract

A scheme based on a W-shaped axicon mirror device for total-internal-reflection fluorescence microscopy (TIRFM) is presented. This approach combines the advantages of higher efficiency compared with traditional TIRFM, adjustable illumination area, and simple switching between wide-field and TIRF imaging modes. TIRF images obtained with this approach are free of shadow artifacts and of interference fringes. Example micrographs of fluorescently labeled polystyrene beads, of Convallaria majalis tissue, and of Propidium-iodide-labeled Chinese hamster ovary cells are shown, and the capabilities of the scheme are discussed.

© 2010 Optical Society of America

Full Article  |  PDF Article

References

  • View by:
  • |
  • |
  • |

  1. J. Pawley, Handbook of Biological Confocal Microscopy, 3rd ed. (Springer, 2006).
    [CrossRef]
  2. D. Axelrod, Methods Cell Biol. 89, 169 (2008).
    [CrossRef]
  3. C. Joo, H. Balci, Y. Ishitsuka, C. Buranachai, and T. Ha, Annu. Rev. Biochem. 77, 51 (2008).
    [CrossRef] [PubMed]
  4. A. L. Stout and D. Axelrod, Appl. Opt. 28, 5237 (1989).
    [CrossRef] [PubMed]
  5. J. N. Forkey, M. E. Quinlan, and Y. E. Goldman, Biophys. J. 89, 1261 (2005).
    [CrossRef] [PubMed]
  6. R. Zoncu, R. M. Perera, R. Sebastian, F. Nakatsu, H. Chen, T. Balla, G. Ayala, D. Toomre, and P. V. DeCamille, Proc. Natl. Acad. Sci. USA 104, 3793 (2007).
    [CrossRef] [PubMed]
  7. D. N. Bowser and B. S. Khakh, Proc. Natl. Acad. Sci. USA 104, 4212 (2007).
    [CrossRef] [PubMed]
  8. G. T. Sincerbox, H. W. Werlich, and B. H. Yung, “Brightfield/darkfield microscope illuminator,” U.S. patent 4,585,315 (April 29, 1986).
  9. M. Endo, Jpn. J. Appl. Phys. 46, 593 (2007).
    [CrossRef]
  10. A. Shoham, R. Vander, and S. G. Lipson, Opt. Lett. 31, 3405 (2006).
    [CrossRef] [PubMed]
  11. P. Alix, J. Winterer, and W. Müller, J. Neurosci. Methods 128, 79 (2003).
    [CrossRef] [PubMed]

2008 (2)

D. Axelrod, Methods Cell Biol. 89, 169 (2008).
[CrossRef]

C. Joo, H. Balci, Y. Ishitsuka, C. Buranachai, and T. Ha, Annu. Rev. Biochem. 77, 51 (2008).
[CrossRef] [PubMed]

2007 (3)

R. Zoncu, R. M. Perera, R. Sebastian, F. Nakatsu, H. Chen, T. Balla, G. Ayala, D. Toomre, and P. V. DeCamille, Proc. Natl. Acad. Sci. USA 104, 3793 (2007).
[CrossRef] [PubMed]

D. N. Bowser and B. S. Khakh, Proc. Natl. Acad. Sci. USA 104, 4212 (2007).
[CrossRef] [PubMed]

M. Endo, Jpn. J. Appl. Phys. 46, 593 (2007).
[CrossRef]

2006 (1)

2005 (1)

J. N. Forkey, M. E. Quinlan, and Y. E. Goldman, Biophys. J. 89, 1261 (2005).
[CrossRef] [PubMed]

2003 (1)

P. Alix, J. Winterer, and W. Müller, J. Neurosci. Methods 128, 79 (2003).
[CrossRef] [PubMed]

1989 (1)

Alix, P.

P. Alix, J. Winterer, and W. Müller, J. Neurosci. Methods 128, 79 (2003).
[CrossRef] [PubMed]

Axelrod, D.

Ayala, G.

R. Zoncu, R. M. Perera, R. Sebastian, F. Nakatsu, H. Chen, T. Balla, G. Ayala, D. Toomre, and P. V. DeCamille, Proc. Natl. Acad. Sci. USA 104, 3793 (2007).
[CrossRef] [PubMed]

Balci, H.

C. Joo, H. Balci, Y. Ishitsuka, C. Buranachai, and T. Ha, Annu. Rev. Biochem. 77, 51 (2008).
[CrossRef] [PubMed]

Balla, T.

R. Zoncu, R. M. Perera, R. Sebastian, F. Nakatsu, H. Chen, T. Balla, G. Ayala, D. Toomre, and P. V. DeCamille, Proc. Natl. Acad. Sci. USA 104, 3793 (2007).
[CrossRef] [PubMed]

Bowser, D. N.

D. N. Bowser and B. S. Khakh, Proc. Natl. Acad. Sci. USA 104, 4212 (2007).
[CrossRef] [PubMed]

Buranachai, C.

C. Joo, H. Balci, Y. Ishitsuka, C. Buranachai, and T. Ha, Annu. Rev. Biochem. 77, 51 (2008).
[CrossRef] [PubMed]

Chen, H.

R. Zoncu, R. M. Perera, R. Sebastian, F. Nakatsu, H. Chen, T. Balla, G. Ayala, D. Toomre, and P. V. DeCamille, Proc. Natl. Acad. Sci. USA 104, 3793 (2007).
[CrossRef] [PubMed]

DeCamille, P. V.

R. Zoncu, R. M. Perera, R. Sebastian, F. Nakatsu, H. Chen, T. Balla, G. Ayala, D. Toomre, and P. V. DeCamille, Proc. Natl. Acad. Sci. USA 104, 3793 (2007).
[CrossRef] [PubMed]

Endo, M.

M. Endo, Jpn. J. Appl. Phys. 46, 593 (2007).
[CrossRef]

Forkey, J. N.

J. N. Forkey, M. E. Quinlan, and Y. E. Goldman, Biophys. J. 89, 1261 (2005).
[CrossRef] [PubMed]

Goldman, Y. E.

J. N. Forkey, M. E. Quinlan, and Y. E. Goldman, Biophys. J. 89, 1261 (2005).
[CrossRef] [PubMed]

Ha, T.

C. Joo, H. Balci, Y. Ishitsuka, C. Buranachai, and T. Ha, Annu. Rev. Biochem. 77, 51 (2008).
[CrossRef] [PubMed]

Ishitsuka, Y.

C. Joo, H. Balci, Y. Ishitsuka, C. Buranachai, and T. Ha, Annu. Rev. Biochem. 77, 51 (2008).
[CrossRef] [PubMed]

Joo, C.

C. Joo, H. Balci, Y. Ishitsuka, C. Buranachai, and T. Ha, Annu. Rev. Biochem. 77, 51 (2008).
[CrossRef] [PubMed]

Khakh, B. S.

D. N. Bowser and B. S. Khakh, Proc. Natl. Acad. Sci. USA 104, 4212 (2007).
[CrossRef] [PubMed]

Lipson, S. G.

Müller, W.

P. Alix, J. Winterer, and W. Müller, J. Neurosci. Methods 128, 79 (2003).
[CrossRef] [PubMed]

Nakatsu, F.

R. Zoncu, R. M. Perera, R. Sebastian, F. Nakatsu, H. Chen, T. Balla, G. Ayala, D. Toomre, and P. V. DeCamille, Proc. Natl. Acad. Sci. USA 104, 3793 (2007).
[CrossRef] [PubMed]

Pawley, J.

J. Pawley, Handbook of Biological Confocal Microscopy, 3rd ed. (Springer, 2006).
[CrossRef]

Perera, R. M.

R. Zoncu, R. M. Perera, R. Sebastian, F. Nakatsu, H. Chen, T. Balla, G. Ayala, D. Toomre, and P. V. DeCamille, Proc. Natl. Acad. Sci. USA 104, 3793 (2007).
[CrossRef] [PubMed]

Quinlan, M. E.

J. N. Forkey, M. E. Quinlan, and Y. E. Goldman, Biophys. J. 89, 1261 (2005).
[CrossRef] [PubMed]

Sebastian, R.

R. Zoncu, R. M. Perera, R. Sebastian, F. Nakatsu, H. Chen, T. Balla, G. Ayala, D. Toomre, and P. V. DeCamille, Proc. Natl. Acad. Sci. USA 104, 3793 (2007).
[CrossRef] [PubMed]

Shoham, A.

Sincerbox, G. T.

G. T. Sincerbox, H. W. Werlich, and B. H. Yung, “Brightfield/darkfield microscope illuminator,” U.S. patent 4,585,315 (April 29, 1986).

Stout, A. L.

Toomre, D.

R. Zoncu, R. M. Perera, R. Sebastian, F. Nakatsu, H. Chen, T. Balla, G. Ayala, D. Toomre, and P. V. DeCamille, Proc. Natl. Acad. Sci. USA 104, 3793 (2007).
[CrossRef] [PubMed]

Vander, R.

Werlich, H. W.

G. T. Sincerbox, H. W. Werlich, and B. H. Yung, “Brightfield/darkfield microscope illuminator,” U.S. patent 4,585,315 (April 29, 1986).

Winterer, J.

P. Alix, J. Winterer, and W. Müller, J. Neurosci. Methods 128, 79 (2003).
[CrossRef] [PubMed]

Yung, B. H.

G. T. Sincerbox, H. W. Werlich, and B. H. Yung, “Brightfield/darkfield microscope illuminator,” U.S. patent 4,585,315 (April 29, 1986).

Zoncu, R.

R. Zoncu, R. M. Perera, R. Sebastian, F. Nakatsu, H. Chen, T. Balla, G. Ayala, D. Toomre, and P. V. DeCamille, Proc. Natl. Acad. Sci. USA 104, 3793 (2007).
[CrossRef] [PubMed]

Annu. Rev. Biochem. (1)

C. Joo, H. Balci, Y. Ishitsuka, C. Buranachai, and T. Ha, Annu. Rev. Biochem. 77, 51 (2008).
[CrossRef] [PubMed]

Appl. Opt. (1)

Biophys. J. (1)

J. N. Forkey, M. E. Quinlan, and Y. E. Goldman, Biophys. J. 89, 1261 (2005).
[CrossRef] [PubMed]

J. Neurosci. Methods (1)

P. Alix, J. Winterer, and W. Müller, J. Neurosci. Methods 128, 79 (2003).
[CrossRef] [PubMed]

Jpn. J. Appl. Phys. (1)

M. Endo, Jpn. J. Appl. Phys. 46, 593 (2007).
[CrossRef]

Methods Cell Biol. (1)

D. Axelrod, Methods Cell Biol. 89, 169 (2008).
[CrossRef]

Opt. Lett. (1)

Proc. Natl. Acad. Sci. USA (2)

R. Zoncu, R. M. Perera, R. Sebastian, F. Nakatsu, H. Chen, T. Balla, G. Ayala, D. Toomre, and P. V. DeCamille, Proc. Natl. Acad. Sci. USA 104, 3793 (2007).
[CrossRef] [PubMed]

D. N. Bowser and B. S. Khakh, Proc. Natl. Acad. Sci. USA 104, 4212 (2007).
[CrossRef] [PubMed]

Other (2)

G. T. Sincerbox, H. W. Werlich, and B. H. Yung, “Brightfield/darkfield microscope illuminator,” U.S. patent 4,585,315 (April 29, 1986).

J. Pawley, Handbook of Biological Confocal Microscopy, 3rd ed. (Springer, 2006).
[CrossRef]

Cited By

OSA participates in CrossRef's Cited-By Linking service. Citing articles from OSA journals and other participating publishers are listed here.

Alert me when this article is cited.


Figures (4)

Fig. 1
Fig. 1

Experimental layout of the TIRF microscopy by using W-shaped axicon mirrors.

Fig. 2
Fig. 2

Comparison of wide-field and TIRF images of 1 μm fluorescence microspheres: (a) wide-field fluorescence image and (b) TIRF image.

Fig. 3
Fig. 3

Comparison of wide-field and TIRF images of Convallaria majalis: (a) wide-field fluorescence image and (b) TIRF image.

Fig. 4
Fig. 4

TIRF images of PI-labeled CHO cells.

Metrics