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Journal of the Optical Society of America A

Journal of the Optical Society of America A

| OPTICS, IMAGE SCIENCE, AND VISION

  • Vol. 13, Iss. 3 — Mar. 1, 1996
  • pp: 479–482

Optical transfer functions for confocal theta fluorescence microscopy

Steffen Lindek and Ernst H. K. Stelzer  »View Author Affiliations


JOSA A, Vol. 13, Issue 3, pp. 479-482 (1996)
http://dx.doi.org/10.1364/JOSAA.13.000479


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Abstract

Optical transfer functions are presented for the confocal theta fluorescence microscope, which uses an orthogonally placed objective lens to detect the fluorescence emission. Therefore the transfer functions do not exhibit cylindrical symmetry about the illumination axis. We show that in an ideal noise-free system, confocal theta microscopy increases the cutoff spatial frequency along the illumination axis by a factor of 3.5 to 3.7 for a numerical aperture of 0.75. In a system with 10% noise the improvement by confocal theta microscopy is even greater, between a factor of 4.1 and 4.4. This explains the improved three-dimensional imaging properties of theta microscopy.

© 1996 Optical Society of America

History
Original Manuscript: June 12, 1995
Revised Manuscript: September 5, 1995
Manuscript Accepted: October 9, 1995
Published: March 1, 1996

Citation
Steffen Lindek and Ernst H. K. Stelzer, "Optical transfer functions for confocal theta fluorescence microscopy," J. Opt. Soc. Am. A 13, 479-482 (1996)
http://www.opticsinfobase.org/josaa/abstract.cfm?URI=josaa-13-3-479


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References

  1. S. Lindek, E. H. K. Stelzer, “Confocal theta microscopy and 4Pi-confocal theta microscopy,” in Three-Dimensional Microscopy: Image Acquisition and Processing, C. J. Cogswell, K. Carlsson, eds., Proc. Soc. Photo-Opt. Instrum. Eng.2184, 188–194 (1994). [CrossRef]
  2. E. H. K. Stelzer, S. Lindek, “Fundamental reduction of the observation volume in far-field light microscopy by detection orthogonal to the illumination axis: confocal theta microscopy,” Opt. Commun. 111, 536–547 (1994). [CrossRef]
  3. S. Lindek, R. Pick, E. H. K. Stelzer, “Confocal theta microscope with three objective lenses,” Rev. Sci. Instrum. 65, 3367–3372 (1994). [CrossRef]
  4. E. H. K. Stelzer, S. Lindek, S. Albrecht, R. Pick, G. Ritter, N. Salmon, R. Stricker, “A new tool for the observation of embryos and other large specimens: confocal theta fluorescence microscopy,” J. Microsc. 179, 1–10 (1995). [CrossRef]
  5. C. J. R. Sheppard, M. Gu, “3-D transfer functions in confocal scanning microscopy,” in Visualization in Biomedical Microscopies, A. Kriete, ed. (Verlag Chemie, Weinheim, Germany, 1992), pp. 251–282.
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  7. B. R. Frieden, “Optical transfer of the three-dimensional object,” J. Opt. Soc. Am. 57, 56–66 (1967). [CrossRef]
  8. C. J. R. Sheppard, M. Gu, “The significance of 3-D transfer functions in confocal scanning microscopy,” J. Microsc. 165, 377–390 (1992). [CrossRef]
  9. M. Gu, T. Tannous, C. J. R. Sheppard, “Improved axial resolution in confocal fluorescence microscopy using annular pupils,” Opt. Commun. 110, 533–539 (1994). [CrossRef]
  10. M. Gu, C. J. R. Sheppard, “Comparison of three-dimensional imaging properties between two-photon and single-photon confocal fluorescence microscopy,” J. Microsc. 177, 128–137 (1995). [CrossRef]
  11. S. Wolfram, mathematica—A System for Doing Mathematics by Computer(Addison-Wesley, Redwood City, Calif., 1991).

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