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Virtual Journal for Biomedical Optics

Virtual Journal for Biomedical Optics


  • Editors: Andrew Dunn and Anthony Durkin
  • Vol. 7, Iss. 12 — Dec. 19, 2012

3D-resolved fluorescence and phosphorescence lifetime imaging using temporal focusing wide-field two-photon excitation

Heejin Choi, Dimitrios S. Tzeranis, Jae Won Cha, Philippe Clémenceau, Sander J. G. de Jong, Lambertus K. van Geest, Joong Ho Moon, Ioannis V. Yannas, and Peter T. C. So  »View Author Affiliations

Optics Express, Vol. 20, Issue 24, pp. 26219-26235 (2012)

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Fluorescence and phosphorescence lifetime imaging are powerful techniques for studying intracellular protein interactions and for diagnosing tissue pathophysiology. While lifetime-resolved microscopy has long been in the repertoire of the biophotonics community, current implementations fall short in terms of simultaneously providing 3D resolution, high throughput, and good tissue penetration. This report describes a new highly efficient lifetime-resolved imaging method that combines temporal focusing wide-field multiphoton excitation and simultaneous acquisition of lifetime information in frequency domain using a nanosecond gated imager from a 3D-resolved plane. This approach is scalable allowing fast volumetric imaging limited only by the available laser peak power. The accuracy and performance of the proposed method is demonstrated in several imaging studies important for understanding peripheral nerve regeneration processes. Most importantly, the parallelism of this approach may enhance the imaging speed of long lifetime processes such as phosphorescence by several orders of magnitude.

© 2012 OSA

OCIS Codes
(110.6880) Imaging systems : Three-dimensional image acquisition
(170.3650) Medical optics and biotechnology : Lifetime-based sensing
(180.2520) Microscopy : Fluorescence microscopy
(180.6900) Microscopy : Three-dimensional microscopy

ToC Category:

Original Manuscript: September 17, 2012
Revised Manuscript: October 26, 2012
Manuscript Accepted: October 27, 2012
Published: November 6, 2012

Virtual Issues
Vol. 7, Iss. 12 Virtual Journal for Biomedical Optics

Heejin Choi, Dimitrios S. Tzeranis, Jae Won Cha, Philippe Clémenceau, Sander J. G. de Jong, Lambertus K. van Geest, Joong Ho Moon, Ioannis V. Yannas, and Peter T. C. So, "3D-resolved fluorescence and phosphorescence lifetime imaging using temporal focusing wide-field two-photon excitation," Opt. Express 20, 26219-26235 (2012)

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