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Optics Express

Optics Express

  • Editor: C. Martijn de Sterke
  • Vol. 19, Iss. 21 — Oct. 10, 2011
  • pp: 20743–20750

Three-dimensional Fluorescence Lifetime Imaging with a Single Plane Illumination Microscope provides an improved Signal to Noise Ratio

Klaus Greger, Manuel J. Neetz, Emmanuel G. Reynaud, and Ernst H.K. Stelzer  »View Author Affiliations

Optics Express, Vol. 19, Issue 21, pp. 20743-20750 (2011)

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We designed a widefield frequency domain Fluorescence Lifetime Imaging Microscopy (FLIM)setup, which is based on a Single Plane Illumination Microscope (SPIM). A SPIM provides an inherent optical sectioning capability and reduces photobleaching compared to conventional widefield and confocal fluorescence microscopes. The lifetime precision of the FLIM was characterized with Rhodamine 6G solutions of different quencher concentrations [KI]. We demonstrate the high spatial resolution of the SPIM-FLIM combination in the intensity domain as well as in the lifetime domain with latex bead samples and multiple recordings of three-dimensional live Madine-Darby Canine Kidney (MDCK) cysts. We estimate that the bleaching rate after 600 images have been recorded is below 5%.

© 2011 OSA

OCIS Codes
(170.2520) Medical optics and biotechnology : Fluorescence microscopy
(170.3650) Medical optics and biotechnology : Lifetime-based sensing
(180.6900) Microscopy : Three-dimensional microscopy

ToC Category:
Medical Optics and Biotechnology

Original Manuscript: July 6, 2011
Revised Manuscript: September 2, 2011
Manuscript Accepted: September 5, 2011
Published: October 4, 2011

Virtual Issues
Vol. 6, Iss. 11 Virtual Journal for Biomedical Optics

Klaus Greger, Manuel J. Neetz, Emmanuel G. Reynaud, and Ernst H.K. Stelzer, "Three-dimensional Fluorescence Lifetime Imaging with a Single Plane Illumination Microscope provides an improved Signal to Noise Ratio," Opt. Express 19, 20743-20750 (2011)

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