| Direct plate-reader measurement of nitric oxide released from hypoxic erythrocytes flowing through a microfluidic device. | |
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MedLine Citation:
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PMID: 20681630 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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The ability to perform a fluorescence-based quantitative determination of a biologically important analyte directly released from mammalian cells using a standard microtiter plate reader to measure wells integrated into a microfluidic device is reported. Specifically, the amount of nitric oxide (NO) released from flowing erythrocytes (ERYs) exposed to a hypoxic buffer is measured using a fluorescein-based probe. The ERYs are pumped through channels in one layer of the poly(dimethylsiloxane) (PDMS) device; as these cells release NO, it flows through a porous polycarbonate membrane to the probe. The device is then placed into a standard microtiter plate reader for measurement, with the entire calibration and analyte determination occurring simultaneously. Using this method, NO release from hypoxic ERYs was determined to be 6.9 +/- 1.8 microM, a significantly increased value in comparison to that from normoxic ERYs of 0.60 +/- 0.04 microM (p < 0.001, n = 4 rabbits). Furthermore, the reproducibility (reported as a %RSD) of measuring fluorescence standards was 3.5%. Detection limits, dynamic range, and optimal membrane pore diameters are also reported. This device enables the use of a standard high-throughput tool (the plate reader) to measure analytes in a microfluidic device, the ability to improve the quantitative determination of a relatively unstable molecule (NO), and the incorporation of a flow component and blood constituent into a system that can be combined with microtiter plate technology. |
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Authors:
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Stephen T Halpin; Dana M Spence |
Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Analytical chemistry Volume: 82 ISSN: 1520-6882 ISO Abbreviation: Anal. Chem. Publication Date: 2010 Sep |
Date Detail:
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Created Date: 2010-08-31 Completed Date: 2011-01-04 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 0370536 Medline TA: Anal Chem Country: United States |
Other Details:
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Languages: eng Pagination: 7492-7 Citation Subset: IM |
Affiliation:
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Department of Chemistry, Michigan State University, East Lansing, Michigan 48824, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Cell Hypoxia Dimethylpolysiloxanes / chemistry Erythrocytes / metabolism* Male Microfluidic Analytical Techniques / instrumentation, methods* Nitric Oxide / metabolism* Rabbits Spectrometry, Fluorescence |
| Chemical | |
Reg. No./Substance:
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0/Dimethylpolysiloxanes; 10102-43-9/Nitric Oxide; 63148-62-9/baysilon |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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