Resealable, optically accessible, PDMS-free fluidic platform for ex vivo interrogation of pancreatic islets.
Resealable, optically accessible, PDMS-free fluidic platform for ex vivo interrogation of pancreatic islets.
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DOI：
10.1039/c6lc01504b
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发表时间：
2017-02-28
期刊：
影响因子：
6.1
通讯作者：
Agarwal A
中科院分区：
文献类型：
--
作者：
Lenguito G;Chaimov D;Weitz JR;Rodriguez-Diaz R;Rawal SA;Tamayo-Garcia A;Caicedo A;Stabler CL;Buchwald P;Agarwal A
我们报告了一个强大的流体平台的设计和制造的惰性塑料材料和微机械加工的功能，促进优化的对流流体输送。该平台被测试用于啮齿动物和人类胰岛的灌注询问、激素的动态分泌、伴随的活细胞成像以及遗传工程胰岛的光遗传学刺激。首先利用葡萄糖刺激的胰岛素分泌和流体动力学的耦合定量流体动力学计算模型来设计对于三维胰岛的完全灌注、分泌的胰岛素的有效收集以及系统体积和相关延迟的最小化是最佳的装置几何形状。然后通过快速原型技术，如微铣削和激光雕刻，将Fluorescent装置制造成两个互锁部件，这些部件由非吸收性和惰性材料制成。最后，使用啮齿动物和人类胰岛测试组件的性能，同时进行多种测定，例如在标准显微镜上进行动态灌注、染色和光遗传学，以及与商业灌注机的集成。对流流体流动的优化设计、生物惰性和非吸收性材料的使用、可逆组装、用于装载和卸载胰岛的手动访问以及与商业成像和流体处理系统的直接集成被证明对于灌注测定是至关重要的，并且特别适合于时间分辨光遗传学研究。由非吸收性材料制成的可逆密封流体平台，其允许3D胰岛培养、灌注测试和时间分辨成像测定
We report the design and fabrication of a robust fluidic platform built out of inert plastic materials and micromachined features that promote optimized convective fluid transport. The platform is tested for perfusion interrogation of rodent and human pancreatic islets, dynamic secretion of hormones, concomitant live-cell imaging, and optogenetic stimulation of genetically engineered islets. A coupled quantitative fluid dynamics computational model of glucose stimulated insulin secretion and fluid dynamics was first utilized to design device geometries that are optimal for complete perfusion of three-dimensional islets, effective collection of secreted insulin, and minimization of system volumes and associated delays. Fluidic devices were then fabricated through rapid prototyping techniques, such as micromilling and laser engraving, as two interlocking parts from materials that are non-absorbent and inert. Finally, the assembly was tested for performance using both rodent and human islets with multiple assays conducted in parallel, such as dynamic perfusion, staining and optogenetics on standard microscopes, as well as for integration with commercial perfusion machines. The optimized design of convective fluid flows, use of bio-inert and non-absorbent materials, reversible assembly, manual access for loading and unloading of islets, and straightforward integration with commercial imaging and fluid handling systems proved to be critical for perfusion assay, and particularly suited for time-resolved optogenetics studies. A reversibly sealing fluidic platform fabricated from non-absorbent materials that permits 3D islet cultures, perfusion testing, and time-resolved imaging assays