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Optics Express

Optics Express

  • Editor: Michael Duncan
  • Vol. 11, Iss. 22 — Nov. 3, 2003
  • pp: 2964–2969

Point spread function of optical microscopes imaging through stratified media

O. Haeberlé, M. Ammar, H. Furukawa, K. Tenjimbayashi, and P. Török  »View Author Affiliations

Optics Express, Vol. 11, Issue 22, pp. 2964-2969 (2003)

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We propose a model for imaging point objects through a dielectric interface or stratified media. The model is applicable to conventional and confocal fluorescence microscopy, with single- or multiphoton excitation. An analytical solution is obtained in the form of readily computable functions. When large mismatches occur in the refractive indices of the media of the objective lens and specimen the illumination and detection point spread functions differ significantly, showing that currently used imaging models may fail to correctly predict imaging properties of optical microscopes.

© 2003 Optical Society of America

OCIS Codes
(050.1960) Diffraction and gratings : Diffraction theory
(180.0180) Microscopy : Microscopy
(260.0260) Physical optics : Physical optics

ToC Category:
Research Papers

Original Manuscript: September 16, 2003
Revised Manuscript: October 27, 2003
Published: November 3, 2003

O. Haeberlé, M. Ammar, H. Furukawa, K. Tenjimbayashi, and P. Török, "The point spread function of optical microscopes imaging through stratified media," Opt. Express 11, 2964-2969 (2003)

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  1. F. Perrin, �??La fluorescence des solutions,�?? Ann. Phys. (Paris) 12, 169-275 (1929)
  2. P. Soleillet, �??Sur les paramètres caractérisant la polarization partielle de la lumière dans les phénomènes de fluorescence,�?? Ann. Phys. (Paris) 12, 23-86 (1929)
  3. D. Axelrod, �??Carbocyanine dye orientation in red cell membrane studied by microscopi fluorescence polarization,�?? Biophys. J. 26, 557-574 (1979) [CrossRef] [PubMed]
  4. I. Gryczynski, H. Malak and J.R. Lakowicz, �??Multiphoton excitation of the DNA stains DAPI and Hoechst,�?? Bioimaging 4, 138-148 (1996) [CrossRef]
  5. P. Török, P.D. Higdon, and T. Wilson, �??On the general properties of polarized light conventional and confocal microscopes,�?? Opt. Commun. 148, 300-315 (1998) [CrossRef]
  6. P.D. Higdon, P. Török, and T. Wilson, �??Imaging properties of high aperture multiphoton fluorescence scanning optical microscopes,�?? J. Microsc. (Oxford) 193, 127-141 (1999) [CrossRef]
  7. P. Török, P.D. Higdon and T. Wilson, �??Theory for confocal and conventional microscopes imaging small dielectric scatterers,�?? J. Mod. Opt. 45, 1681-1698 (1998) [CrossRef]
  8. C.J.R. Sheppard and P. Török, �??An electromagnetic theory of imaging in fluorescence microscopy, and imaging in polarization fluorescence microscopy,�?? Bioimaging 5, 205-218 (1997) [CrossRef]
  9. P. Török and P. Varga, �??Electromagnetic diffraction of light focused through a stratified medium,�?? Appl. Opt. 36, 2305-2312 (1997) [CrossRef] [PubMed]
  10. P. Török, �??Propagation of electromagnetic dipole waves through dielectric interfaces,�?? Opt. Lett. 25, 1463-1465 (2000) [CrossRef]
  11. Note that there was a typo in Eq. (6) and Eq. (14c) defining Ydet in Ref. 10
  12. D. Minsky, �??Memoir on Inventing the Confocal Scanning Microscope,�?? Scanning 10, 128-138 (1988) [CrossRef]
  13. S. Hell and E.H.K. Stelzer, �??Fundamental improvement of resolution with a 4-Pi-confocal microscope using two-photon excitation,�?? Opt. Comm. 93, 277-281 (1992) [CrossRef]
  14. E. H. K. Stelzer and 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]
  15. O. Haeberlé et al., �??Multiple-objective microscopy with three-dimensional resolution near 100 nm and a long working distance,�?? Opt. Lett. 26, 1684-1686 (2001) [CrossRef]
  16. O. Haeberlé, �??Focusing of light through a stratified medium: a practical approach for computing microscope point spread functions. Part I: Conventional microscopy,�?? Opt. Commun. 216, 55-63 (2003) [CrossRef]

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