Integration of multiple components in polystyrene-based microfluidic devices part II: cellular analysis.
Integration of multiple components in polystyrene-based microfluidic devices part II: cellular analysis.
复制标题
DOI：
10.1039/c2an36171j
复制
发表时间：
2013-01-07
期刊：
影响因子：
--
通讯作者：
Spence DM
中科院分区：
文献类型：
--
作者：
Anderson KB;Halpin ST;Johnson AS;Martin RS;Spence DM
在本系列的第二部分中，描述了聚苯乙烯（PS）设备用于微流控细胞分析的使用，各种细胞类型和检测策略被用来确定通常与细胞相关的三种基本分析。具体来说，使用集成电化学传感或光学测量与标准多孔板读取器，细胞摄取，生产，或重要的细胞分析物的释放是在基于ps的设备上确定的。一个实验涉及荧光测量一氧化氮（NO）产生内皮细胞系后，刺激ATP。结果是一氧化氮的产生增加了四倍（与对照组相比），这种基于受体的一氧化氮产生机制验证了固定在PS底物上后细胞受体的维持。在Ca2+离子载体A20317刺激后，通过光学测定Ca2+进入内皮细胞，也证明了监测细胞摄取的能力。结果表明，与对照组（17%）相比，离子载体存在时钙的吸收量显著增加（42%）（p < 0.05）。最后，从多巴胺能细胞系（pc12细胞）中释放儿茶酚胺的电化学监测，电极被嵌入到基于ps的装置中。与使用PDMS相比，pc12细胞在PS装置上的粘附性更好。钾刺激导致114±11µM儿茶酚胺的释放，比暴露于抑制剂（利血平，20±2µM儿茶酚胺）的细胞释放量显著增加（p < 0.05）。成功测量多种分析物的能力，以不同的方式从不同的研究细胞中产生，表明PS可能是微流控装置制造的有用材料，特别是考虑到PS增强的细胞粘附性，其增强的刚性/自动化适应性，以及它能够研究更广泛的分析物，甚至具有高度疏水性的分析物。
In Part II of this series describing the use of polystyrene (PS) devices for microfluidic-based cellular assays, various cellular types and detection strategies are employed to determine three fundamental assays often associated with cells. Specifically, using either integrated electrochemical sensing or optical measurements with a standard multi-well plate reader, cellular uptake, production, or release of important cellular analytes are determined on a PS-based device. One experiment involved the fluorescence measurement of nitric oxide (NO) produced within an endothelial cell line following stimulation with ATP. The result was a four-fold increase in NO production (as compared to a control), with this receptor-based mechanism of NO production verifying the maintenance of cell receptors following immobilization onto the PS substrate. The ability to monitor cellular uptake was also demonstrated by optical determination of Ca2+ into endothelial cells following stimulation with the Ca2+ ionophore A20317. The result was a significant increase (42%) in the calcium uptake in the presence of the ionophore, as compared to a control (17%) (p < 0.05). Finally, the release of catecholamines from a dopaminergic cell line (PC 12 cells) was electrochemically monitored, with the electrodes being embedded into the PS-based device. The PC 12 cells had better adherence on the PS devices, as compared to use of PDMS. Potassium-stimulation resulted in the release of 114 ± 11 µM catecholamines, a significant increase (p < 0.05) over the release from cells that had been exposed to an inhibitor (reserpine, 20 ± 2 µM of catecholamines). The ability to successfully measure multiple analytes, generated in different means from various cells under investigation, suggests that PS may be a useful material for microfluidic device fabrication, especially considering the enhanced cell adhesion to PS, its enhanced rigidity/amenability to automation, and its ability to enable a wider range of analytes to be investigated, even analytes with a high degree of hydrophobicity.
登录
查看更多内容
DOI：
10.1152/ajpheart.1996.271.6.h2717
发表时间：
1996-12-01
影响因子：
4.8
作者：
Sprague, RS;Ellsworth, ML;Lonigro, AJ
通讯作者：
Lonigro, AJ
影响因子：
3
作者：
Li, MW;Spence, DM;Martin, RS
通讯作者：
Martin, RS
影响因子：
4.3
作者：
Detwiler, David A.;Dobes, Nicholas C.;Allbritton, Nancy L.
通讯作者：
Allbritton, Nancy L.
影响因子：
6.1
作者：
Regehr KJ;Domenech M;Koepsel JT;Carver KC;Ellison-Zelski SJ;Murphy WL;Schuler LA;Alarid ET;Beebe DJ
通讯作者：
Beebe DJ
影响因子：
6.1
作者：
Genes, Luiza I.;Tolan, Nicole V.;Spence, Dana M.
通讯作者：
Spence, Dana M.