An integrated microfluidic device for monitoring changes in nitric oxide production in single T-lymphocyte (Jurkat) cells.
An integrated microfluidic device for monitoring changes in nitric oxide production in single T-lymphocyte (Jurkat) cells.
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DOI：
10.1021/ac401665u
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发表时间：
2013-11-05
影响因子：
7.4
通讯作者：
Culbertson, Christopher T.
中科院分区：
文献类型：
--
作者：
Metto, Eve C.;Evans, Karsten;Barney, Patrick;Culbertson, Anne H.;Gunasekara, Dulan B.;Caruso, Giuseppe;Huvey, Matthew K.;Fracassi da Silva, Jose Alberto;Lunte, Susan M.;Culbertson, Christopher T.
为了更好地了解癌症和神经退行性疾病中的细胞异质性，人们已经将相当多的注意力集中在单细胞的分析上。虽然微流控装置在单细胞分析方面有一些优势，但很少有论文实际证明这些装置能够监测受干扰生物系统中的化学变化。本文描述了一种新的微流体通道歧管，它将细胞运输，裂解，注射，电泳分离和荧光检测集成到一个设备中，使其能够以自动化方式以10个细胞/分钟的速度分析单个细胞。该系统使用荧光标记物4-氨基-5-甲氨基-2',7'-二乙酸二氟荧光素（DAF-FM DA）检测单个t淋巴细胞（Jurkat细胞）中一氧化氮（NO）的产生。细胞也用6-羧基荧光素二醋酸酯（6-CFDA）作为内标进行标记。对照细胞的NO生成与使用脂多糖（LPS）刺激的细胞的NO生成进行了比较，已知脂多糖在免疫型细胞中引起诱导型一氧化氮合酶（iNOS）的表达。对一群细胞的电泳图进行统计分析表明，诱导细胞中NO的产生增加了两倍。这些结果很好地与最近发表的一氧化氮的大量细胞分析相比较。
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.
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影响因子：
3.9
作者：
Lai, Hsuan-Hong;Quinto-Su, Pedro A.;Allbritton, Nancy L.
通讯作者：
Allbritton, Nancy L.
DOI：
10.1039/c2an16211c
发表时间：
2012-07-07
期刊：
The Analyst
影响因子：
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作者：
Cecala C;Sweedler JV
通讯作者：
Sweedler JV
影响因子：
4.1
作者：
Greif, Dominik;Galla, Lukas;Anselmetti, Dario
通讯作者：
Anselmetti, Dario
DOI：
10.1385/1-59745-076-6:203
发表时间：
2006-01-01
期刊：
MICROCHIP CAPILLARY ELECTROPHORESIS: METHODS AND PROTOCOLS
影响因子：
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作者：
Culbertson, Christopher T.
通讯作者：
Culbertson, Christopher T.
DOI：
10.1007/978-1-61779-445-2_28
发表时间：
2012-01-01
期刊：
AMINO ACID ANALYSIS: METHODS AND PROTOCOLS
影响因子：
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作者：
Liu, Yi-Ming;Zhao, Shulin
通讯作者：
Zhao, Shulin