Dynamic monitoring of glucagon secretion from living cells on a microfluidic chip.
Dynamic monitoring of glucagon secretion from living cells on a microfluidic chip.
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DOI：
10.1007/s00216-012-5755-7
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发表时间：
2012-03
影响因子：
4.3
通讯作者：
Kennedy, Robert T.
中科院分区：
文献类型：
--
作者：
Shackman, Jonathan G.;Reid, Kendra R.;Dugan, Colleen E.;Kennedy, Robert T.
建立了一种基于微流控技术的毛细管电泳法(CEIA)，用于在线监测朗格汉斯胰岛高血糖素的分泌。在该装置中，含有活的胰岛的细胞室被注入了含有高或低葡萄糖浓度的缓冲液。通过芯片上的单独通道通过电渗透连续采样灌流液。将灌流液与异硫氰酸荧光素标记的胰高血糖素(FITC-GUP)和抗胰高血糖素单抗在线混合。为了最大限度地减少样品稀释，通过允许仅通过扩散添加试剂，使样品灌流液与试剂的芯片上混合比率最大化。每隔6个S，将反应混合物注入1.5 cm的分离通道，在700V cm−1的电场下分离游离的荧光素-胰高血糖素和游离的荧光素-胰高血糖素抗体复合物。当灌流液中的葡萄糖浓度从15 mM降至1 mM时，定量监测各组胰岛中胰高血糖素分泌的变化，显示出在阶跃变化中胰高血糖素分泌的脉冲。这个高度自动化的系统应该能够研究胰升糖素的调节及其在糖尿病和肥胖症中的潜在作用。该方法还进一步展示了在微流控系统中用于监测细胞功能的快速CEIA的潜力。
A rapid microfluidic based capillary electrophoresis immunoassay (CEIA) was developed for on-line monitoring of glucagon secretion from pancreatic islets of Langerhans. In the device, a cell chamber containing living islets was perfused with buffers containing either high or low glucose concentration. Perfusate was continuously sampled by electroosmosis through a separate channel on the chip. The perfusate was mixed on-line with fluorescein isothiocyanate-labeled glucagon (FITC-glucagon) and monoclonal anti-glucagon antibody. To minimize sample dilution, the on-chip mixing ratio of sampled perfusate to reagents was maximized by allowing reagents to only be added by diffusion. Every 6 s the reaction mixture was injected onto a 1.5 cm separation channel where free FITC-glucagon and the FITC-glucagon-antibody complex were separated under an electric field of 700 V cm−1. The immunoassay had a detection limit of 1 nM. Groups of islets were quantitatively monitored for changes in glucagon secretion as the glucose concentration was decreased from 15 to 1 mM in the perfusate revealing a pulse of glucagon secretion during a step change. The highly automated system should be enable studies of the regulation of glucagon and its potential role in diabetes and obesity. The method also further demonstrates the potential of rapid CEIA on microfluidic systems for monitoring cellular function.
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影响因子：
4.1
作者：
Guillo, Christelle;Roper, Michael G.
通讯作者：
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影响因子：
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作者：
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7.4
作者：
Dishinger, John F.;Reid, Kendra R.;Kennedy, Robert T.
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作者：
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通讯作者：
Ramsey, JM
影响因子：
7.4
作者：
Edwards, JL;Kennedy, RT
通讯作者：
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