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Optics Express

Optics Express

  • Editor: C. Martijn de Sterke
  • Vol. 20, Iss. 4 — Feb. 13, 2012
  • pp: 3744–3752

Scattered light fluorescence microscopy in three dimensions

Giulia Ghielmetti and Christof M. Aegerter  »View Author Affiliations

Optics Express, Vol. 20, Issue 4, pp. 3744-3752 (2012)

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Recently, we have proposed a method to image fluorescent structures behind turbid layers at diffraction limited resolution using wave-front shaping and the memory effect. However, this was limited to a raster scanning of the wave-front shaped focus to a two dimensional plane. In applications, it can however be of great importance to be able to scan a three dimensional volume. Here we show that this can be implemented in the same setup. This is achieved by the addition of a parabolic phase shift to the shaped wave-front. Via the memory effect, this phase shift leads to a shift of the interference based focus in the z-direction, thus opening the possibility of three dimensional imaging using scattered light fluorescence microscopy. Here, we show an example of such a three dimensional image of fluorescent nano-beads taken behind a turbid layer more than 10 mean free paths thick. Finally, we discuss the differences of the scanning in the z-direction with that in the x–y plane and the corresponding possibilities and limitations of the technique.

© 2012 OSA

OCIS Codes
(030.6140) Coherence and statistical optics : Speckle
(110.0180) Imaging systems : Microscopy
(110.7050) Imaging systems : Turbid media
(290.4210) Scattering : Multiple scattering

ToC Category:

Original Manuscript: December 7, 2011
Revised Manuscript: January 23, 2012
Manuscript Accepted: January 26, 2012
Published: January 31, 2012

Virtual Issues
Vol. 7, Iss. 4 Virtual Journal for Biomedical Optics

Giulia Ghielmetti and Christof M. Aegerter, "Scattered light fluorescence microscopy in three dimensions," Opt. Express 20, 3744-3752 (2012)

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