期刊
ANALYTICAL CHEMISTRY
卷 85, 期 21, 页码 10188-10195出版社
AMER CHEMICAL SOC
DOI: 10.1021/ac401665u
关键词
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资金
- NIH [R21NS061202]
- Terry Johnson Cancer Center, Kansas State University
- DOD ASSURE [CHE-1004991]
- University of Catania, Italy
- American Heart Association
- Fundacao de Amparoa Pesquisa do Estado de Sao Paulo, FAPESP [2010/01046-6]
- Division Of Chemistry
- Direct For Mathematical & Physical Scien [1004991] Funding Source: National Science Foundation
- Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP) [10/01046-6] Funding Source: FAPESP
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 2-fold increase in NO production in the induced cells. These results compare nicely to a recently published bulk cell analysis of NO.
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