Adewola Adeola F, Wang Yong, Harvat Tricia, Eddington David T, Lee Dongyoung, Oberholzer Jose
Department of Surgery.
J Vis Exp. 2010 Jan 26(35):1649. doi: 10.3791/1649.
A microfluidic islet perifusion device was developed for the assessment of dynamic insulin secretion of multiple islets and simultaneous fluorescence imaging of calcium influx and mitochondrial potential changes. The device consists of three layers: first layer contains an array of microscale wells (500 mum diameter and 150 mum depth) that help to immobilize the islets while exposed to flow and maximize the exposed surface area of the islets; the second layer contains a circular perifusion chamber (3 mm deep, 7 mm diameter); and the third layer contains an inlet-mixing channel that fans out before injection into the perifusion chamber (2 mm in width, 19 mm in length, and 500 mum in height) for optimizing the mixing efficiency prior to entering the perifusion chamber. The creation of various glucose gradients including a linear, bell shape, and square shapes also can be created in the microfluidic perifusion network and is demonstrated.
开发了一种微流控胰岛灌流装置，用于评估多个胰岛的动态胰岛素分泌以及同时对钙内流和线粒体电位变化进行荧光成像。该装置由三层组成：第一层包含一系列微尺度孔（直径500μm，深度150μm），有助于在胰岛暴露于流动时固定胰岛，并使胰岛的暴露表面积最大化；第二层包含一个圆形灌流室（深3mm，直径7mm）；第三层包含一个入口混合通道，该通道在注入灌流室之前呈扇形展开（宽2mm，长19mm，高500μm），用于在进入灌流室之前优化混合效率。还可以在微流控灌流网络中创建包括线性、钟形和方形在内的各种葡萄糖梯度，并进行了展示。
