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Optics Letters

Optics Letters

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  • Vol. 26, Iss. 11 — Jun. 1, 2001
  • pp: 828–830

Three-dimensional resolution enhancement in fluorescence microscopy by harmonic excitation

J. T. Frohn, H. F. Knapp, and A. Stemmer  »View Author Affiliations


Optics Letters, Vol. 26, Issue 11, pp. 828-830 (2001)
http://dx.doi.org/10.1364/OL.26.000828


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Abstract

A method for increasing lateral as well as axial resolution in fluorescence microscopy is presented. A passband with a high cutoff frequency throughout reciprocal space can be achieved by illumination of the object with spatially harmonic excitation patterns generated by the interference of two collimated laser beams. Theoretical calculations show an almost isotropic point-spread function with a FWHM near 100 nm.

© 2001 Optical Society of America

OCIS Codes
(070.0070) Fourier optics and signal processing : Fourier optics and signal processing
(100.3020) Image processing : Image reconstruction-restoration
(100.6640) Image processing : Superresolution
(180.0180) Microscopy : Microscopy
(180.2520) Microscopy : Fluorescence microscopy

Citation
J. T. Frohn, H. F. Knapp, and A. Stemmer, "Three-dimensional resolution enhancement in fluorescence microscopy by harmonic excitation," Opt. Lett. 26, 828-830 (2001)
http://www.opticsinfobase.org/ol/abstract.cfm?URI=ol-26-11-828


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References

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  2. T. Wilson, in Handbook of Biological Confocal Microscopy, J. B. Pawley, ed. (Plenum, New York, 1995), p. 167.
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  9. Absolute fringe position D1 of the first image can easily be calculated from the measured fluorescence images because the original passband and the shifted copies overlap. This fact simplifies a practical 3D HELM setup because the beam deflection units are not required to maintain a fixed phase relationship during operation.
  10. A. Erhardt, G. Zinser, D. Komitowski, and J. Bille, Appl. Opt. 24, 194 (1998).
  11. The choice of a cosine bell apodization results in image points with negative intensities. One normalizes the traces in Figs. – by setting the most negative value to zero.
  12. The spatial frequency of excitation u equals the difference of the propagation vectors p1 and p2 of the interfering waves. Therefore a rotation of both p1 and p2 about an axis parallel to p1–p2 leaves the pattern unaffected. By an appropriate rotation of p1 and p2, any desired value of u can be realized by interference of two plane waves whose propagation vectors reside in lower and upper half-spaces, respectively, relative to the object plane.

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