Quantitative monitoring of insulin secretion from single islets of Langerhans in parallel on a microfluidic chip.
Quantitative monitoring of insulin secretion from single islets of Langerhans in parallel on a microfluidic chip.
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DOI：
10.1021/ac900109t
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发表时间：
2009-04-15
影响因子：
7.4
通讯作者：
Kennedy, Robert T.
中科院分区：
文献类型：
--
作者：
Dishinger, John F.;Reid, Kendra R.;Kennedy, Robert T.
胰岛素从胰岛释放的定量对于糖尿病研究是有意义的。典型的胰岛素分泌实验是使用离线技术进行的，这些技术昂贵、缓慢、通量低，并且需要多个胰岛。我们已经开发了一种微流控装置，用于并行地高通量、自动化和在线监测单个胰岛的胰岛素分泌。该芯片由15个通道网络组成，每个通道网络能够灌注单个胰岛，并将来自每个胰岛的灌注液与异硫氰酸荧光素标记的胰岛素和抗胰岛素抗体在线混合，用于竞争性免疫测定。将得到的连续反应流周期性地注入到平行的电泳通道上，在电泳通道中分离混合物。所得迹线用于定量从胰岛释放的相对胰岛素。在所有15个通道上以10秒的间隔进行系列免疫测定，对应于每小时5400次免疫测定，以产生捕获单个胰岛分泌动力学的时间分辨的胰岛素释放曲线。该芯片用于证明游离脂肪酸诱导的胰岛中的脂毒性消除了脉动胰岛素分泌。
Quantification of insulin release from pancreatic islets of Langerhans is of interest for diabetes research. Typical insulin secretion experiments are performed using offline techniques that are expensive, slow, have low-throughput, and require multiple islets. We have developed a microfluidic device for high-throughput, automated, and online monitoring of insulin secretion from individual islets in parallel. This chip consists of 15 channel networks each capable of superfusing a single islet and mixing superfusate from each islet online with fluorescein isothiocyanate-labeled insulin and anti-insulin antibody for a competitive immunoassay. The resulting continuous reaction streams are periodically injected onto parallel electrophoresis channels where the mixtures are separated. The resulting traces are used to quantify relative insulin released from islets. Serial immunoassays were performed at 10 s intervals on all 15 channels, corresponding to 5400 immunoassays per hour, to create temporally resolved insulin release profiles that captured single islet secretion dynamics. The chip was used to demonstrate that free fatty acid induced lipotoxicity in islets eliminates pulsatile insulin secretion.
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影响因子：
7.7
作者：
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通讯作者：
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DOI：
10.1152/ajpendo.1998.274.5.e796
发表时间：
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影响因子：
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