Institute for Biomaterials and Biomedical Engineering, University of Toronto, 164 College St, Toronto, ON M5S 3G9, Canada.
Integr Biol (Camb). 2013 Aug;5(8):1014-25. doi: 10.1039/c3ib40104a.
The potential benefits of using new technologies such as microfluidics for life science applications are exciting, but it is critical to understand and document potential biases imposed by these technologies on the observed results. Here, we report the first study of genome-level effects on cells manipulated by digital microfluidics. These effects were evaluated using a broad suite of tools: cell-based stress sensors for heat shock activation, single-cell COMET assays to probe changes in DNA integrity, and DNA microarrays and qPCR to evaluate changes in genetic expression. The results lead to two key observations. First, most DMF operating conditions tested, including those that are commonly used in the literature, result in negligible cell-stress or genome-level effects. Second, for DMF devices operated at high driving frequency (18 kHz) and with large driving electrodes (10 mm × 10 mm), there are significant damage to DNA integrity and differential genomic regulation. We hypothesize that these effects are caused by droplet heating. We recommend that for DMF applications involving mammalian cells that driving frequencies be kept low (≤ 10 kHz) and electrode sizes be kept small (≤ 5 mm) to avoid detrimental effects.
使用微流控等新技术应用于生命科学具有令人兴奋的潜在益处，但理解和记录这些技术对观察结果产生的潜在偏差至关重要。在这里，我们报告了首次对数字微流控处理的细胞进行全基因组水平影响的研究。这些影响使用了一系列广泛的工具进行评估：用于热休克激活的基于细胞的应激传感器、用于探测 DNA 完整性变化的单细胞彗星测定法、以及用于评估基因表达变化的 DNA 微阵列和 qPCR。研究结果得出了两个关键观察结果。首先，测试的大多数数字微流控操作条件，包括文献中常用的条件，都不会导致细胞应激或全基因组水平的影响。其次，对于在高驱动频率（18 kHz）和大驱动电极（10 mm×10 mm）下运行的数字微流控设备，会导致 DNA 完整性严重受损和基因组调控的差异。我们假设这些影响是由液滴加热引起的。我们建议，对于涉及哺乳动物细胞的数字微流控应用，应保持低驱动频率（≤10 kHz）和小电极尺寸（≤5 mm），以避免有害影响。
