Fluorescence Monitoring of ATP-Stimulated, Endothelium-Derived Nitric Oxide Production in Channels of a Poly(dimethylsiloxane)-Based Microfluidic Device
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- 作者：Teresa D'Amico Oblak ; Paul Root ; and Dana M. Spence
- 刊名：Analytical Chemistry
- 出版年：2006
- 出版时间：May 1, 2006
- 年：2006
- 卷：78
- 期：9
- 页码：3193 - 3197
- 全文大小：148K
- 年卷期：v.78,no.9(May 1, 2006)
- ISSN：1520-6882
文摘
Intracellular nitric oxide (NO) production in a microfluidicendothelium is detected using fluorescence microscopy.Bovine pulmonary artery endothelial cells (bPAECs) wereloaded with the fluorescence probe diaminodifluorofluorescein diacetate (DAF-FM DA), and the subsequentfluorescent DAF-FM DA/NO adduct was measured. Solutions of bradykinin, a well-known stimulus of endothelium-derived NO, activated nitric oxide synthase (NOS)in the immobilized bPAECs. This activation was inhibitedusing L-nitro arginine methyl ester (L-NAME), a competitive inhibitor of NOS. Importantly, the NO production wasalso stimulated with adenosine triphosphate (ATP) usingconcentrations as low as 1 
M. Previous reports onstimulating NO production using an immobilized endothelium in microfluidic channels were limited by therequirement of ATP concentrations of at least 100 M, avalue that is not physiologically relevant. The ability tomonitor NO production with ATP concentrations that aresimilar to in vivo levels of ATP in the microcirculationrepresents a major advance in the use of microfluidictechnology as an in vitro model of the microcirculation.
M. Previous reports onstimulating NO production using an immobilized endothelium in microfluidic channels were limited by therequirement of ATP concentrations of at least 100 M, avalue that is not physiologically relevant. The ability tomonitor NO production with ATP concentrations that aresimilar to in vivo levels of ATP in the microcirculationrepresents a major advance in the use of microfluidictechnology as an in vitro model of the microcirculation.