Tkachenko Eugene, Gutierrez Edgar, Ginsberg Mark H, Groisman Alex
Department of Medicine, University of California, San Diego, 9500 Gilman Drive, MC 0726, La Jolla, CA 92093, USA.
Lab Chip. 2009 Apr 21;9(8):1085-95. doi: 10.1039/b812184b. Epub 2009 Feb 6.
We have built and characterized a magnetic clamp for reversible sealing of PDMS microfluidic chips against cover glasses with cell cultures and a microfluidic chip for experiments on shear stress response of endothelial cells. The magnetic clamp exerts a reproducible uniform pressure on the microfluidic chip, achieving fast and reliable sealing for liquid pressures up to 40 kPa inside the chip with <10% deformations of microchannels and minimal variations of the substrate shear stress in perfusion flow. The microfluidic chip has 8 test regions with the substrate shear stress varying by a factor of 2 between each region, thus covering a 128-fold range from low venous to arterial. The perfusion is driven by differential pressure, which makes it possible to create pulsatile flows mimicking pulsing in the vasculature. The setup is tested by 15-40 hours perfusions over endothelial monolayers with shear stress in the range of 0.07-9 dyn/cm(2). Excellent cell viability at all shear stresses and alignment of cells along the flow at high shear stresses are repeatedly observed. A scratch wound healing assay under a shear flow is demonstrated and cell migration velocities are measured. Transfection of cells with a fluorescent protein is performed, and migrating fluorescent cells are imaged at a high resolution under shear flow in real time. The magnetic clamp can be closed with minimal mechanical perturbation to cells on the substrate and used with a variety of microfluidic chips for experiments with adherent and non-adherent cells.
我们构建并表征了一种磁性夹具,用于将带有细胞培养物的聚二甲基硅氧烷(PDMS)微流控芯片与盖玻片进行可逆密封,还构建了一种用于内皮细胞剪切应力响应实验的微流控芯片。该磁性夹具能在微流控芯片上施加可重复的均匀压力,对于芯片内部高达40 kPa的液体压力,能实现快速可靠的密封,微通道变形小于10%,且在灌注流中基底剪切应力变化极小。该微流控芯片有8个测试区域,每个区域之间的基底剪切应力变化系数为2,从而覆盖了从低静脉到动脉的128倍范围。灌注由压差驱动,这使得模拟脉管系统中的脉动产生脉动流成为可能。该装置在0.07 - 9 dyn/cm²的剪切应力下对内皮细胞单层进行了15 - 40小时的灌注测试。在所有剪切应力下均观察到优异的细胞活力,并且在高剪切应力下细胞沿流动方向排列。展示了在剪切流下的划痕伤口愈合试验并测量了细胞迁移速度。对细胞进行了荧光蛋白转染,并在剪切流下实时对迁移的荧光细胞进行了高分辨率成像。该磁性夹具可以在对基底上的细胞产生最小机械扰动的情况下关闭,并可与各种微流控芯片一起用于贴壁细胞和非贴壁细胞的实验。