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用于实时活力和化学反应评估的微流控装置中单细胞的阻抗和光学检测。

Impedimetric and optical interrogation of single cells in a microfluidic device for real-time viability and chemical response assessment.

作者信息

James Conrad D, Reuel Nigel, Lee Eunice S, Davalos Rafael V, Mani Seethambal S, Carroll-Portillo Amanda, Rebeil Roberto, Martino Anthony, Apblett Christopher A

机构信息

Sandia National Laboratories, PO Box 5800, Albuquerque, MS 87185-1080, United States.

出版信息

Biosens Bioelectron. 2008 Jan 18;23(6):845-51. doi: 10.1016/j.bios.2007.08.022. Epub 2007 Sep 6.

Abstract

We report here a non-invasive, reversible method for interrogating single cells in a microfluidic flow-through system. Impedance spectroscopy of cells held at a micron-sized pore under negative pressure is demonstrated and used to determine the presence and viability of the captured cell. The cell capture pore is optimized for electrical response and mechanical interfacing to a cell using a deposited layer of parylene. Changes in the mechanical interface between the cell and the chip due to chemical exposure or environmental changes can also be assessed. Here, we monitored the change in adhesion/spreading of RAW264.7 macrophages in response to the immune stimulant lipopolysaccharide (LPS). This method enables selective, reversible, and quantitative long-term impedance measurements on single cells. The fully sealed electrofluidic assembly is compatible with long-term cell culturing, and could be modified to incorporate single cell lysis and subsequent intracellular separation and analysis.

摘要

我们在此报告一种用于在微流控流通系统中检测单细胞的非侵入性、可逆方法。展示了在负压下对处于微米级孔隙中的细胞进行阻抗谱分析,并用于确定捕获细胞的存在和活力。细胞捕获孔通过聚对二甲苯沉积层针对电响应和与细胞的机械接口进行了优化。由于化学暴露或环境变化导致的细胞与芯片之间机械界面的变化也可以进行评估。在此,我们监测了RAW264.7巨噬细胞对免疫刺激物脂多糖(LPS)的粘附/铺展变化。该方法能够对单细胞进行选择性、可逆和定量的长期阻抗测量。完全密封的电流体组件与长期细胞培养兼容,并且可以进行修改以纳入单细胞裂解以及随后的细胞内分离和分析。

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