Mollet Mike, Godoy-Silva Ruben, Berdugo Claudia, Chalmers Jeffrey J
Department of Chemical and Biomolecular Engineering, The Ohio State University, 125 Koffolt Laboratories, 140 W 19th Ave, Columbus, Ohio 43210, USA.
Biotechnol Bioeng. 2008 Jun 1;100(2):260-72. doi: 10.1002/bit.21762.
Fluorescence activated cell sorting, FACS, is a widely used method to sort subpopulations of cells to high purities. To achieve relatively high sorting speeds, FACS instruments operate by forcing suspended cells to flow in a single file line through a laser(s) beam(s). Subsequently, this flow stream breaks up into individual drops which can be charged and deflected into multiple collection streams. Previous work by Ma et al. (2002) and Mollet et al. (2007; Biotechnol Bioeng 98:772-788) indicates that subjecting cells to hydrodynamic forces consisting of both high extensional and shear components in micro-channels results in significant cell damage. Using the fluid dynamics software FLUENT, computer simulations of typical fluid flow through the nozzle of a BD FACSVantage indicate that hydrodynamic forces, quantified using the scalar parameter energy dissipation rate, are similar in the FACS nozzle to levels reported to create significant cell damage in micro-channels. Experimental studies in the FACSVantage, operated under the same conditions as the simulations confirmed significant cell damage in two cell lines, Chinese Hamster Ovary cells (CHO) and THP1, a human acute monocytic leukemia cell line.
荧光激活细胞分选术(FACS)是一种广泛应用的将细胞亚群分选至高纯度的方法。为实现相对较高的分选速度,FACS仪器通过迫使悬浮细胞以单列形式流过一束或多束激光束来进行操作。随后,该流动流会分裂成单个液滴,这些液滴可被充电并偏转到多个收集流中。Ma等人(2002年)以及Mollet等人(2007年;《生物技术与生物工程》98:772 - 788)之前的研究表明,使细胞在微通道中受到由高拉伸和剪切成分组成的流体动力作用会导致显著的细胞损伤。使用流体动力学软件FLUENT,对典型流体流经BD FACSVantage喷嘴的计算机模拟表明,使用标量参数能量耗散率量化的流体动力在FACS喷嘴中与据报道在微通道中造成显著细胞损伤的水平相似。在与模拟相同条件下运行的FACSVantage中进行的实验研究证实,两种细胞系,即中国仓鼠卵巢细胞(CHO)和人急性单核细胞白血病细胞系THP1,均出现了显著的细胞损伤。