Polanco Edward R, Western Nicholas, Zangle Thomas A
Department of Chemical Engineering, University of Utah.
Department of Chemical Engineering, University of Utah; Huntsman Cancer Institute, University of Utah;
J Vis Exp. 2018 Sep 10(139):58296. doi: 10.3791/58296.
The use of microfluidic devices has emerged as a defining tool for biomedical applications. When combined with modern microscopy techniques, these devices can be implemented as part of a robust platform capable of making simultaneous complementary measurements. The primary challenge created by the combination of these two techniques is the mismatch in refractive index between the materials traditionally used to make microfluidic devices and the aqueous solutions typically used in biomedicine. This mismatch can create optical artifacts near the channel or device edges. One solution is to reduce the refractive index of the material used to fabricate the device by using a fluorinated polymer such as MY133-V2000 whose refractive index is similar to that of water (n = 1.33). Here, the construction of a microfluidic device made out of MY133-V2000 using soft lithography techniques is demonstrated, using O2 plasma in conjunction with an acrylic holder to increase the adhesion between the MY133-V2000 fabricated device and the polydimethylsiloxane (PDMS) substrate. The device is then tested by incubating it filled with cell culture media for 24 h to demonstrate the ability of the device to maintain cell culture conditions during the course of a typical imaging experiment. Finally, quantitative phase microscopy (QPM) is used to measure the distribution of mass within the live adherent cells in the microchannel. This way, the increased precision, enabled by fabricating the device from a low index of refraction polymer such as MY133-V2000 in lieu of traditional soft lithography materials such as PDMS, is demonstrated. Overall, this approach for fabricating microfluidic devices can be readily integrated into existing soft lithography workflows in order to reduce optical artifacts and increase measurement precision.
微流控设备的使用已成为生物医学应用中的一项关键工具。当与现代显微镜技术相结合时,这些设备可作为一个强大平台的一部分来实现,该平台能够进行同步互补测量。这两种技术结合所带来的主要挑战是,传统上用于制造微流控设备的材料与生物医学中通常使用的水溶液之间存在折射率不匹配的问题。这种不匹配会在通道或设备边缘附近产生光学伪像。一种解决方案是通过使用诸如MY133 - V2000之类的氟化聚合物来降低用于制造设备的材料的折射率,该聚合物的折射率与水的折射率相似(n = 1.33)。在此,展示了使用软光刻技术由MY133 - V2000制成的微流控设备的构建过程,通过结合使用O2等离子体和丙烯酸支架来增强由MY133 - V2000制成的设备与聚二甲基硅氧烷(PDMS)基板之间的附着力。然后,将充满细胞培养基的该设备孵育24小时进行测试,以证明该设备在典型成像实验过程中维持细胞培养条件的能力。最后,使用定量相显微镜(QPM)来测量微通道中活贴壁细胞内的质量分布。通过这种方式,展示了用诸如MY133 - V2000之类的低折射率聚合物代替诸如PDMS之类的传统软光刻材料制造设备所带来的更高精度。总体而言,这种制造微流控设备的方法可以很容易地集成到现有的软光刻工作流程中,以减少光学伪像并提高测量精度。