Dubay R, Fiering J, Darling E M
Draper, Cambridge, Massachusetts 02139, USA.
Biomicrofluidics. 2020 Aug 3;14(4):044110. doi: 10.1063/5.0017770. eCollection 2020 Jul.
Label-free microfluidic-based cell sorters leverage innate differences among cells (e.g., size and stiffness), to separate one cell type from another. This sorting step is crucial for many cell-based applications. Polystyrene-based microparticles (MPs) are the current gold standard for calibrating flow-based cell sorters and analyzers; however, the deformation behavior of these rigid materials is drastically different from that of living cells. Given this discrepancy in stiffness, an alternative calibration particle that better reflects cell elasticity is needed for the optimization of new and existing microfluidic devices. Here, we describe the fabrication of cell-like, mechanically tunable MPs and demonstrate their utility in quantifying differences in inertial displacement within a microfluidic constriction device as a function of particle elastic modulus, for the first time. Monodisperse, fluorescent, cell-like microparticles that replicate the size and modulus of living cells were fabricated from polyacrylamide within a microfluidic droplet generator and characterized via optical and atomic force microscopy. Trajectories of our cell-like MPs were mapped within the constriction device to predict where living cells of similar size/modulus would move. Calibration of the device with our MPs showed that inertial displacement depends on both particle size and modulus, with large/soft MPs migrating further toward the channel centerline than small/stiff MPs. The mapped trajectories also indicated that MP modulus contributed proportionally more to particle displacement than size, for the physiologically relevant ranges tested. The large shift in focusing position quantified here emphasizes the need for physiologically relevant, deformable MPs for calibrating and optimizing microfluidic separation platforms.
基于无标记微流控的细胞分选仪利用细胞之间的固有差异(如大小和硬度)来将一种细胞类型与另一种细胞类型分离。这一分选步骤对于许多基于细胞的应用至关重要。基于聚苯乙烯的微粒子(MPs)是当前校准基于流式细胞分选仪和分析仪的金标准;然而,这些刚性材料的变形行为与活细胞的变形行为截然不同。鉴于这种硬度差异,需要一种能更好反映细胞弹性的替代校准粒子来优化新的和现有的微流控设备。在此,我们描述了类细胞、机械可调MPs的制造,并首次展示了它们在量化微流控收缩装置内惯性位移差异方面的效用,该差异是作为粒子弹性模量的函数。在微流控液滴发生器内由聚丙烯酰胺制备了单分散、荧光、类细胞微粒子,其复制了活细胞的大小和模量,并通过光学和原子力显微镜进行了表征。我们的类细胞MPs的轨迹在收缩装置内被绘制,以预测类似大小/模量的活细胞会移动到何处。用我们的MPs对该装置进行校准表明,惯性位移取决于粒子大小和模量,大/软的MPs比小/硬的MPs向通道中心线迁移得更远。绘制的轨迹还表明,在所测试的生理相关范围内,MP模量对粒子位移的贡献比大小成比例地更大。此处量化的聚焦位置的大偏移强调了需要生理相关的、可变形的MPs来校准和优化微流控分离平台。