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在电动微流控系统中诱导和抑制细胞裂解。

Induction and suppression of cell lysis in an electrokinetic microfluidic system.

机构信息

Department of Chemical Engineering, Michigan Technological University, Houghton, Michigan, USA.

出版信息

Electrophoresis. 2022 Jun;43(12):1322-1336. doi: 10.1002/elps.202100310. Epub 2022 Apr 26.

DOI:10.1002/elps.202100310
PMID:35306692
Abstract

The ability to strategically induce or suppress cell lysis is critical for many cellular-level diagnostic and therapeutic applications conducted within electrokinetic microfluidic platforms. The chemical and structural integrity of sub-cellular components is important when inducing cell lysis. However, metal electrodes and electrolytes participate in undesirable electrochemical reactions that alter solution composition and potentially damage protein, RNA, and DNA integrity within device microenvironments. For many biomedical applications, cell viability must be maintained even when device-imposed cell-stressing stimuli (e.g., electrochemical reaction byproducts) are present. In this work, we explored a novel and tunable method to accurately induce or suppress device-imposed artifacts on human red blood cell (RBC) lysis in non-uniform AC electric fields. For precise tunability, a dielectric hafnium oxide (HfO ) layer was used to prevent electron transfer between the electrodes and the electric double layer and thus reduce harmful electrochemical reactions. Additionally, a low concentration of Triton X-100 surfactant was explored as a tool to stabilize cell membrane integrity. The extent of hemolysis was studied as a function of time, electrode configuration (T-shaped and star-shaped), cell position, applied non-uniform AC electric field, with uncoated and HfO coated electrodes (50 nm), and absence and presence of Triton X-100 (70 µM). Tangible outcomes include a parametric analysis relying upon literature and this work to design, tune, and operate electrokinetic microdevices to intentionally induce or suppress cellular lysis without altering intracellular components. Implications are that devices can be engineered to leverage or minimize device-imposed biological artefacts extending the versatility and utility of electrokinetic diagnostics.

摘要

在基于电动微流控平台进行的许多细胞水平诊断和治疗应用中,战略性地诱导或抑制细胞裂解的能力至关重要。在诱导细胞裂解时,亚细胞成分的化学和结构完整性很重要。然而,金属电极和电解质参与了不理想的电化学反应,这些反应改变了溶液的组成,并可能破坏设备微环境中蛋白质、RNA 和 DNA 的完整性。对于许多生物医学应用,即使存在设备施加的细胞应激刺激(例如,电化学反应副产物),也必须保持细胞活力。在这项工作中,我们探索了一种新颖且可调谐的方法,以精确诱导或抑制非均匀交流电场中对人红细胞(RBC)裂解的设备施加的人工制品。为了实现精确的调谐性,使用介电氧化铪(HfO )层来防止电极和双电层之间的电子转移,从而减少有害的电化学反应。此外,还探索了低浓度的 Triton X-100 表面活性剂作为稳定细胞膜完整性的工具。溶血程度随时间、电极配置(T 形和星形)、细胞位置、施加的非均匀交流电场、未涂覆和涂覆 HfO 的电极(50nm)以及是否存在 Triton X-100(70µM)而变化。有形的结果包括依赖文献和这项工作进行参数分析,以设计、调整和操作电动微器件,有意诱导或抑制细胞裂解而不改变细胞内成分。这意味着可以设计设备来利用或最小化设备施加的生物人工制品,从而扩展电动诊断的多功能性和实用性。

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