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Proceedings Paper

Extracellular oxygen concentration mapping with a confocal multiphoton laser scanning microscope and TCSPC card
Author(s): Neveen A. Hosny; David A. Lee; Martin M. Knight
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Paper 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.

Paper Details

Date Published: 26 February 2010
PDF: 6 pages
Proc. SPIE 7569, Multiphoton Microscopy in the Biomedical Sciences X, 756932 (26 February 2010); doi: 10.1117/12.842281
Show Author Affiliations
Neveen A. Hosny, Queen Mary Univ. of London (United Kingdom)
David A. Lee, Queen Mary Univ. of London (United Kingdom)
Martin M. Knight, Queen Mary Univ. of London (United Kingdom)

Published in SPIE Proceedings Vol. 7569:
Multiphoton Microscopy in the Biomedical Sciences X
Ammasi Periasamy; Peter T. C. So; Karsten König, Editor(s)

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