1 March 2009 Calibration and validation of an optical sensor for intracellular oxygen measurements
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J. of Biomedical Optics, 14(2), 020506 (2009). doi:10.1117/1.3116714
Abstract
Calibration of fluorescent optical sensors for accurate, quantitative intracellular measurements in vivo suffers from lack of a representative medium that appropriately simulates the molecular complexity of the cytosol. We present a novel protocol for accurate intracellular oxygen sensing via fluorescence lifetime imaging microscopy (FLIM) using cell lysate-FLIM measurements to correct the in vitro calibration of a fluorescent oxygen sensor, and we describe electron paramagnetic resonance (EPR) validation studies. Lysate-FLIM studies provided biochemical information, while EPR provided a "gold standard" for intracellular oxygen estimation. Oxygen levels were evaluated in living human normal squamous and adenocarcinoma esophageal epithelial cells, and good agreement was observed between oxygen levels derived from the optical protocol and EPR. The proposed protocol introduces the concept of a living cell line as a reference for estimating unknown oxygen levels in other cell lines and accounts for high degrees of variability between different cell lines.
Dhruv Sud, Mary-Ann Mycek, "Calibration and validation of an optical sensor for intracellular oxygen measurements," Journal of Biomedical Optics 14(2), 020506 (1 March 2009). http://dx.doi.org/10.1117/1.3116714
Submission: Received ; Accepted
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KEYWORDS
Oxygen

Calibration

Fluorescence lifetime imaging

Luminescence

Optical sensors

Particles

Sensors

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