Abstract

The condenser is one of the main components in most transmitted light compound microscopes. In this Letter, we show that such a condenser can be replaced by a programmable LED array to achieve greater imaging flexibility and functionality. Without mechanically scanning the sample or changing the microscope setup, the proposed approach can be used for dark-field imaging, bright-field imaging, microscopy sectioning, and digital refocusing. Images of a starfish embryo were acquired by using such an approach for demonstration.

© 2011 Optical Society of America

Full Article  |  PDF Article

References

  • View by:
  • |
  • |
  • |

  1. J. James, Light Microscopic Techniques in Biology and Medicine (Springer, 1976).
    [CrossRef]
  2. M. G. L. Gustafsson, Proc. Natl. Acad. Sci. USA 102, 13081 (2005).
    [CrossRef] [PubMed]
  3. V. Poher, H. Zhang, G. Kennedy, C. Griffin, S. Oddos, E. Gu, D. Elson, M. Girkin, P. French, and M. Dawson, Opt. Express 15, 11196 (2007).
    [CrossRef] [PubMed]
  4. J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. H. K. Stelzer, Science 305, 1007 (2004).
    [CrossRef] [PubMed]
  5. J. Wu, G. Zheng, Z. Li, and C. Yang, Opt. Lett. 36, 2179(2011).
    [CrossRef] [PubMed]
  6. F. O. Fahrbach, P. Simon, and A. Rohrbach, Nat. Photon. 4, 780 (2010).
    [CrossRef]
  7. V. Bormuth, J. Howard, and E. Schaffer, J. Microsc. 226, 1 (2007).
    [CrossRef] [PubMed]
  8. V. Murthy, T. F. Sato, D. F. Albeanu, E. Soucy, and M. Meister, PLoS ONE 3 (2008).
    [PubMed]
  9. G. C. Cox, Optical Imaging Techniques in Cell Biology (CRC/Taylor & Francis, Boca Raton, 2007).
  10. G. Zheng, X. Cui, and C. Yang, in Proceedings of the National Academy of Sciences of India, Section A (Physical Sciences) (2010), Vol. 107, p. 9043.
  11. P. J. Shaw, D. A. Agard, Y. Hiraoka, and J. W. Sedat, Biophys. J. 55, 101 (1989).
    [CrossRef] [PubMed]
  12. G. Zheng, S. A. Lee, S. Yang, and C. Yang, Lab Chip 10, 3125 (2010).
    [CrossRef] [PubMed]
  13. A. Barty, K. Nugent, D. Paganin, and A. Roberts, Opt. Lett. 23, 817 (1998).
    [CrossRef]

2011 (1)

2010 (2)

F. O. Fahrbach, P. Simon, and A. Rohrbach, Nat. Photon. 4, 780 (2010).
[CrossRef]

G. Zheng, S. A. Lee, S. Yang, and C. Yang, Lab Chip 10, 3125 (2010).
[CrossRef] [PubMed]

2008 (1)

V. Murthy, T. F. Sato, D. F. Albeanu, E. Soucy, and M. Meister, PLoS ONE 3 (2008).
[PubMed]

2007 (2)

2005 (1)

M. G. L. Gustafsson, Proc. Natl. Acad. Sci. USA 102, 13081 (2005).
[CrossRef] [PubMed]

2004 (1)

J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. H. K. Stelzer, Science 305, 1007 (2004).
[CrossRef] [PubMed]

1998 (1)

1989 (1)

P. J. Shaw, D. A. Agard, Y. Hiraoka, and J. W. Sedat, Biophys. J. 55, 101 (1989).
[CrossRef] [PubMed]

Agard, D. A.

P. J. Shaw, D. A. Agard, Y. Hiraoka, and J. W. Sedat, Biophys. J. 55, 101 (1989).
[CrossRef] [PubMed]

Albeanu, D. F.

V. Murthy, T. F. Sato, D. F. Albeanu, E. Soucy, and M. Meister, PLoS ONE 3 (2008).
[PubMed]

Barty, A.

Bormuth, V.

V. Bormuth, J. Howard, and E. Schaffer, J. Microsc. 226, 1 (2007).
[CrossRef] [PubMed]

Cox, G. C.

G. C. Cox, Optical Imaging Techniques in Cell Biology (CRC/Taylor & Francis, Boca Raton, 2007).

Cui, X.

G. Zheng, X. Cui, and C. Yang, in Proceedings of the National Academy of Sciences of India, Section A (Physical Sciences) (2010), Vol. 107, p. 9043.

Dawson, M.

Del Bene, F.

J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. H. K. Stelzer, Science 305, 1007 (2004).
[CrossRef] [PubMed]

Elson, D.

Fahrbach, F. O.

F. O. Fahrbach, P. Simon, and A. Rohrbach, Nat. Photon. 4, 780 (2010).
[CrossRef]

French, P.

Girkin, M.

Griffin, C.

Gu, E.

Gustafsson, M. G. L.

M. G. L. Gustafsson, Proc. Natl. Acad. Sci. USA 102, 13081 (2005).
[CrossRef] [PubMed]

Hiraoka, Y.

P. J. Shaw, D. A. Agard, Y. Hiraoka, and J. W. Sedat, Biophys. J. 55, 101 (1989).
[CrossRef] [PubMed]

Howard, J.

V. Bormuth, J. Howard, and E. Schaffer, J. Microsc. 226, 1 (2007).
[CrossRef] [PubMed]

Huisken, J.

J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. H. K. Stelzer, Science 305, 1007 (2004).
[CrossRef] [PubMed]

James, J.

J. James, Light Microscopic Techniques in Biology and Medicine (Springer, 1976).
[CrossRef]

Kennedy, G.

Lee, S. A.

G. Zheng, S. A. Lee, S. Yang, and C. Yang, Lab Chip 10, 3125 (2010).
[CrossRef] [PubMed]

Li, Z.

Meister, M.

V. Murthy, T. F. Sato, D. F. Albeanu, E. Soucy, and M. Meister, PLoS ONE 3 (2008).
[PubMed]

Murthy, V.

V. Murthy, T. F. Sato, D. F. Albeanu, E. Soucy, and M. Meister, PLoS ONE 3 (2008).
[PubMed]

Nugent, K.

Oddos, S.

Paganin, D.

Poher, V.

Roberts, A.

Rohrbach, A.

F. O. Fahrbach, P. Simon, and A. Rohrbach, Nat. Photon. 4, 780 (2010).
[CrossRef]

Sato, T. F.

V. Murthy, T. F. Sato, D. F. Albeanu, E. Soucy, and M. Meister, PLoS ONE 3 (2008).
[PubMed]

Schaffer, E.

V. Bormuth, J. Howard, and E. Schaffer, J. Microsc. 226, 1 (2007).
[CrossRef] [PubMed]

Sedat, J. W.

P. J. Shaw, D. A. Agard, Y. Hiraoka, and J. W. Sedat, Biophys. J. 55, 101 (1989).
[CrossRef] [PubMed]

Shaw, P. J.

P. J. Shaw, D. A. Agard, Y. Hiraoka, and J. W. Sedat, Biophys. J. 55, 101 (1989).
[CrossRef] [PubMed]

Simon, P.

F. O. Fahrbach, P. Simon, and A. Rohrbach, Nat. Photon. 4, 780 (2010).
[CrossRef]

Soucy, E.

V. Murthy, T. F. Sato, D. F. Albeanu, E. Soucy, and M. Meister, PLoS ONE 3 (2008).
[PubMed]

Stelzer, E. H. K.

J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. H. K. Stelzer, Science 305, 1007 (2004).
[CrossRef] [PubMed]

Swoger, J.

J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. H. K. Stelzer, Science 305, 1007 (2004).
[CrossRef] [PubMed]

Wittbrodt, J.

J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. H. K. Stelzer, Science 305, 1007 (2004).
[CrossRef] [PubMed]

Wu, J.

Yang, C.

J. Wu, G. Zheng, Z. Li, and C. Yang, Opt. Lett. 36, 2179(2011).
[CrossRef] [PubMed]

G. Zheng, S. A. Lee, S. Yang, and C. Yang, Lab Chip 10, 3125 (2010).
[CrossRef] [PubMed]

G. Zheng, X. Cui, and C. Yang, in Proceedings of the National Academy of Sciences of India, Section A (Physical Sciences) (2010), Vol. 107, p. 9043.

Yang, S.

G. Zheng, S. A. Lee, S. Yang, and C. Yang, Lab Chip 10, 3125 (2010).
[CrossRef] [PubMed]

Zhang, H.

Zheng, G.

J. Wu, G. Zheng, Z. Li, and C. Yang, Opt. Lett. 36, 2179(2011).
[CrossRef] [PubMed]

G. Zheng, S. A. Lee, S. Yang, and C. Yang, Lab Chip 10, 3125 (2010).
[CrossRef] [PubMed]

G. Zheng, X. Cui, and C. Yang, in Proceedings of the National Academy of Sciences of India, Section A (Physical Sciences) (2010), Vol. 107, p. 9043.

Biophys. J. (1)

P. J. Shaw, D. A. Agard, Y. Hiraoka, and J. W. Sedat, Biophys. J. 55, 101 (1989).
[CrossRef] [PubMed]

J. Microsc. (1)

V. Bormuth, J. Howard, and E. Schaffer, J. Microsc. 226, 1 (2007).
[CrossRef] [PubMed]

Lab Chip (1)

G. Zheng, S. A. Lee, S. Yang, and C. Yang, Lab Chip 10, 3125 (2010).
[CrossRef] [PubMed]

Nat. Photon. (1)

F. O. Fahrbach, P. Simon, and A. Rohrbach, Nat. Photon. 4, 780 (2010).
[CrossRef]

Opt. Express (1)

Opt. Lett. (2)

PLoS ONE (1)

V. Murthy, T. F. Sato, D. F. Albeanu, E. Soucy, and M. Meister, PLoS ONE 3 (2008).
[PubMed]

Proc. Natl. Acad. Sci. USA (1)

M. G. L. Gustafsson, Proc. Natl. Acad. Sci. USA 102, 13081 (2005).
[CrossRef] [PubMed]

Science (1)

J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. H. K. Stelzer, Science 305, 1007 (2004).
[CrossRef] [PubMed]

Other (3)

J. James, Light Microscopic Techniques in Biology and Medicine (Springer, 1976).
[CrossRef]

G. C. Cox, Optical Imaging Techniques in Cell Biology (CRC/Taylor & Francis, Boca Raton, 2007).

G. Zheng, X. Cui, and C. Yang, in Proceedings of the National Academy of Sciences of India, Section A (Physical Sciences) (2010), Vol. 107, p. 9043.

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

Proposed illumination scheme. (a) The optical condenser is replaced by a programmable LED array. No lens is placed between the LED array and the sample stage. (b) One LED is turned on for illumination. (c) The LEDs in central part are turned on for the bright-field imaging. (d) The LEDs at the edge are turned on for dark-field imaging. (e) The actual LED array prototype used in our experiment. (f) A simple ray-trace diagram showing different z-planes result in different image shifts (also see Eq. (2)).

Fig. 2
Fig. 2

(a) Bright-field and (b) dark-field images of a starfish embryo captured using the proposed illumination scheme.

Fig. 3
Fig. 3

(a1–a8) Based on the captured images for different individual LEDs, we can digitally refocus the starfish embryo into different depths. No mechanical scanning is involved in this process. (b) The image taken by the conventional Köhler illumination. We use a halogen lamp as the light source with an illumination NA of 0.3 (matching to 0.3 NA objective lens).

Fig. 4
Fig. 4

The images of a 0.5 μm bead by using (a) the proposed method, (b) Köhler illumination. (c) The line traces of (a) and (b).

Equations (2)

Equations on this page are rendered with MathJax. Learn more.

r / r 2 + H 2 ,
s x ( h ) = x i h / H , s y ( h ) = y i h / H ,

Metrics