@inbook{a001d47970e746c5a1e1b5ba4b4c317a,
title = "Extracellular oxygen concentration mapping with confocal multiphoton laser scanning microscope and TCSPC card",
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.",
keywords = "Chondrocytes, oxygen gradients, ruthenium, extracellular, fluorescence lifetime, two-photon microscopy, FLIM, ARTICULAR CHONDROCYTES, IN-VITRO, COMPLEXES, CONSUMPTION, TENSION, PROBES, CELLS, PH",
author = "Neveen Hosny and Lee, {David A.} and Knight, {Martin M.}",
year = "2010",
month = feb,
day = "26",
doi = "10.1117/12.842281",
language = "English",
isbn = "9780819479655",
series = "Proceedings of SPIE",
publisher = "SPIE - INT SOC OPTICAL ENGINEERING",
editor = "A Periasamy and PTC So and K Konig",
booktitle = "MULTIPHOTON MICROSCOPY IN THE BIOMEDICAL SCIENCES X",
note = "Conference on Multiphoton Microscopy in the Biomedical Sciences X ; Conference date: 24-01-2010 Through 26-01-2010",
}