| Calibration and validation of an optical sensor for intracellular oxygen measurements. | |
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MedLine Citation:
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PMID: 19405711 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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. |
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Authors:
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Dhruv Sud; Mary-Ann Mycek |
Publication Detail:
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Type: Evaluation Studies; Letter; Research Support, N.I.H., Extramural; Validation Studies |
Journal Detail:
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Title: Journal of biomedical optics Volume: 14 ISSN: 1083-3668 ISO Abbreviation: J Biomed Opt Publication Date: 2009 Mar-Apr |
Date Detail:
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Created Date: 2009-05-01 Completed Date: 2009-07-15 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9605853 Medline TA: J Biomed Opt Country: United States |
Other Details:
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Languages: eng Pagination: 020506 Citation Subset: IM |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Algorithms Calibration Cells, Cultured / metabolism* Electron Spin Resonance Spectroscopy / instrumentation*, standards* Optical Devices / standards* Oximetry / instrumentation*, standards* Oxygen / analysis* Reproducibility of Results Sensitivity and Specificity |
| Grant Support | |
ID/Acronym/Agency:
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CA-114542/CA/NCI NIH HHS |
| Chemical | |
Reg. No./Substance:
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7782-44-7/Oxygen |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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