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Applied Optics

Applied Optics

APPLICATIONS-CENTERED RESEARCH IN OPTICS

  • Editor: James C. Wyant
  • Vol. 46, Iss. 29 — Oct. 10, 2007
  • pp: 7237–7243

Wide-field depth-sectioning fluorescence microscopy using projector-generated patterned illumination

Serafin Delica and Carlo Mar Blanca  »View Author Affiliations


Applied Optics, Vol. 46, Issue 29, pp. 7237-7243 (2007)
http://dx.doi.org/10.1364/AO.46.007237


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Abstract

We present a simple and cost-effective wide-field, depth-sectioning, fluorescence microscope utilizing a commercial multimedia projector to generate excitation patterns on the sample. Highly resolved optical sections of fluorescent pollen grains at 1.9 μm axial resolution are constructed using the structured illumination technique. This requires grid excitation patterns to be scanned across the sample, which is straightforwardly implemented by creating slideshows of gratings at different phases, projecting them onto the sample, and synchronizing camera acquisition with slide transition. In addition to rapid dynamic pattern generation, the projector provides high illumination power and spectral excitation selectivity. We exploit these properties by imaging mouse neural cells in cultures multistained with Alexa 488 and Cy3. The spectral and structural neural information is effectively resolved in three dimensions. The flexibility and commercial availability of this light source is envisioned to open multidimensional imaging to a broader user base.

© 2007 Optical Society of America

OCIS Codes
(100.3010) Image processing : Image reconstruction techniques
(180.2520) Microscopy : Fluorescence microscopy
(180.6900) Microscopy : Three-dimensional microscopy

ToC Category:
Microscopy

History
Original Manuscript: July 2, 2007
Manuscript Accepted: August 13, 2007
Published: October 5, 2007

Virtual Issues
Vol. 2, Iss. 11 Virtual Journal for Biomedical Optics

Citation
Serafin Delica and Carlo Mar Blanca, "Wide-field depth-sectioning fluorescence microscopy using projector-generated patterned illumination," Appl. Opt. 46, 7237-7243 (2007)
http://www.opticsinfobase.org/ao/abstract.cfm?URI=ao-46-29-7237


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References

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