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    An Integrated Microfluidic Device for Monitoring Changes in Nitric Oxide Production in Single T-Lymphocyte (Jurkat) Cells

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    Metto_AnalChem_2013.pdf (1.919Mb)
    Issue Date
    2013-11-05
    Author
    Metto, Eve C.
    Evans, Karsten
    Barney, Patrick
    Culbertson, Anne H.
    Gunasekara, Dulan B.
    Caruso, Giuseppe
    Hulvey, Matthew K.
    da Silva, José A. F.
    Lunte, Susan M.
    Culbertson, Christopher T.
    Publisher
    American Chemical Society
    Type
    Article
    Article Version
    Scholarly/refereed, author accepted manuscript
    Rights
    Copyright © 2013 American Chemical Society
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    Abstract
    A considerable amount of attention has been focused on the analysis of single cells in an effort to better understand cell heterogeneity in cancer and neurodegenerative diseases. Although microfluidic devices have several advantages for single cell analysis, few papers have actually demonstrated the ability of these devices to monitor chemical changes in perturbed biological systems. In this paper, a new microfluidic channel manifold is described that integrates cell transport, lysis, injection, electrophoretic separation, and fluorescence detection into a single device, making it possible to analyze individual cells at a rate of 10 cells/min in an automated fashion. The system was employed to measure nitric oxide (NO) production in single T-lymphocytes (Jurkat cells) using a fluorescent marker, 4-amino-5-methylamino-2',7'-difluorofluorescein diacetate (DAF-FM DA). The cells were also labeled with 6-carboxyfluorescein diacetate (6-CFDA) as an internal standard. The NO production by control cells was compared to that of cells stimulated using lipopolysaccharide (LPS), which is known to cause the expression of inducible nitric oxide synthase (iNOS) in immune-type cells. Statistical analysis of the resulting electropherograms from a population of cells indicated a twofold increase in NO production in the induced cells. These results compare nicely to a recently published bulk cell analysis of NO.
    URI
    http://hdl.handle.net/1808/23584
    DOI
    https://doi.org/10.1021/ac401665u
    Collections
    • Chemistry Scholarly Works [600]
    Citation
    Metto, E. C., Evans, K., Barney, P., Culbertson, A. H., Gunasekara, D. B., Caruso, G., … Culbertson, C. T. (2013). An Integrated Microfluidic Device for Monitoring Changes in Nitric Oxide Production in Single T-Lymphocyte (Jurkat) Cells. Analytical Chemistry, 85(21), 10188–10195. http://doi.org/10.1021/ac401665u

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    Contact KU ScholarWorks
    785-864-8983
    KU Libraries
    1425 Jayhawk Blvd
    Lawrence, KS 66045
    785-864-8983

    KU Libraries
    1425 Jayhawk Blvd
    Lawrence, KS 66045
    Image Credits
     

     

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