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Proceedings Article

Extracellular oxygen concentration mapping with a confocal multiphoton laser scanning microscope and TCSPC card

[+] Author Affiliations
Neveen A. Hosny, David A. Lee, Martin M. Knight

Queen Mary Univ. of London (United Kingdom)

Proc. SPIE 7569, Multiphoton Microscopy in the Biomedical Sciences X, 756932 (February 26, 2010); doi:10.1117/12.842281
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From Conference Volume 7569

  • Multiphoton Microscopy in the Biomedical Sciences X
  • Ammasi Periasamy; Peter T. C. So; Karsten König
  • San Francisco, California | January 23, 2010

abstract

Extracellular oxygen concentrations influence cell metabolism and tissue function. Fluorescence Lifetime Imaging Microscopy (FLIM) offers a non-invasive method for quantifying local oxygen concentrations. However, existing methods show limited spatial resolution and/or require custom made systems. This study describes a new optimised approach for quantitative extracellular oxygen detection, providing an off-the-shelf system with high spatial resolution and an improved lifetime determination over previous techniques, while avoiding systematic photon pile-up. Fluorescence lifetime detection of an oxygen sensitive fluorescent dye, tris(2,2'-bipyridyl)ruthenium(II) chloride hexahydrate [Ru(bipy)3]2+, was measured using a Becker&Hickl time-correlated single photon counting (TCSPC) card with excitation provided by a multi-photon laser. This technique was able to identify a subpopulation of isolated chondrocyte cells, seeded in three-dimensional agarose gel, displaying a significant spatial oxygen gradient. Thus this technique provides a powerful tool for quantifying spatial oxygen gradients within three-dimensional cellular models.

© (2010) COPYRIGHT SPIE--The International Society for Optical Engineering. Downloading of the abstract is permitted for personal use only.
Citation

Neveen A. Hosny ; David A. Lee and Martin M. Knight
"Extracellular oxygen concentration mapping with a confocal multiphoton laser scanning microscope and TCSPC card", Proc. SPIE 7569, Multiphoton Microscopy in the Biomedical Sciences X, 756932 (February 26, 2010); doi:10.1117/12.842281; http://dx.doi.org/10.1117/12.842281


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