Passively operated microfluidic device for stimulation and secretion sampling of single pancreatic islets.
Passively operated microfluidic device for stimulation and secretion sampling of single pancreatic islets.
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DOI：
10.1021/ac201598b
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发表时间：
2011-09-15
影响因子：
7.4
通讯作者：
Easley CJ
中科院分区：
文献类型：
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作者：
Godwin LA;Pilkerton ME;Deal KS;Wanders D;Judd RL;Easley CJ
设计了一种被动操作的聚二甲基硅氧烷（PDMS）微流控装置，用于平行采集8个小鼠胰岛的激素分泌物。使用单个手持式注射器并利用微通道的固有流体阻力（R采样= 700 ± 20 kPa s mm-3，37 °C）实现流量控制。将基础（3 mM）或刺激（11 mM）葡萄糖水平应用于胰岛，使用改良的储液器将刺激时间（tstim）最小化至15 ± 2 s。采用酶联免疫吸附试验（ELISA）进行后采样分析，我们测量了统计学上相等的1小时胰岛素分泌水平（1.26 ± 0.26和6.55± 1.00 pg胰岛-1 min-1，基础和刺激; 62个胰岛）与标准批量采样方法相比（1.01± 0.224和6.04 ± 1.53 pg胰岛-1 min-1，基础和刺激; 200个胰岛）。重要的是，微流体平台揭示了关于单胰岛变异性的新信息。用共聚焦反射显微镜测量胰岛体积显示，胰岛素分泌与胰岛体积的相关性有限，这表明组织内的细胞结构和旁分泌信号传导具有更重要的作用。与使用注射泵或电渗流控制的其他方法相比，这种方法在易用性和设备一次性使用方面具有显着优势，减轻了非专家的负担。
A passively operated polydimethylsiloxane (PDMS) microfluidic device was designed for sampling of hormone secretions from eight individual murine pancreatic islets in parallel. Flow control was achieved using a single hand-held syringe and by exploiting inherent fluidic resistances of the microchannels (Rsampling = 700 ± 20 kPa s mm−3 at 37 °C). Basal (3 mM) or stimulatory (11 mM) glucose levels were applied to islets, with stimulation timing (tstim) minimized to 15 ± 2 s using modified reservoirs. Using enzyme-linked immunosorbent assays (ELISA) for postsampling analyses, we measured statistically equal levels of 1 h insulin secretion (1.26 ± 0.26 and 6.55± 1.00 pg islet−1 min−1, basal and stimulated; 62 islets) compared to standard, bulk sampling methods (1.01± 0.224 and 6.04 ± 1.53 pg islet−1 min−1, basal and stimulated; 200 islets). Importantly, the microfluidic platform revealed novel information on single-islet variability. Islet volume measurements with confocal reflectance microscopy revealed that insulin secretion had only limited correlation to islet volume, suggesting a more significant role for cellular architecture and paracrine signaling within the tissue. Compared to other methods using syringe pumps or electroosmotic flow control, this approach provides significant advantages in ease-of-use and device disposability, easing the burden on nonexperts.