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实时 QCM-D 监测不同细胞形态试剂对细胞反应的影响。

Real-time QCM-D monitoring of cellular responses to different cytomorphic agents.

机构信息

Department of Chemical Engineering, McGill University, Montreal, Quebec, Canada.

出版信息

Biosens Bioelectron. 2011 Mar 15;26(7):3207-12. doi: 10.1016/j.bios.2010.12.027. Epub 2010 Dec 23.

Abstract

Quartz crystal microbalance with dissipation monitoring (QCM-D) is used for real-time in situ detection of cytoskeletal changes in live primary endothelial cells in response to different cytomorphic agents; namely, the surfactant Triton-X 100 (TX-100) and bacterial lipopolysaccharide (LPS). Reproducible dissipation versus frequency (Df) plots provide unique signatures of the interactions between endothelial cells and cytomorphic agents. While the QCM-D response for TX-100 can be described in two steps (changes in the osmotic pressure of the medium prior to observing the expected cell lysis), LPS results in a different single-phase signal. A complementary analysis is carried out to evaluate the possible competitive effects of TX-100 and LPS through the QCM-D response to BAEC stress by analyzing the Df plots obtained. Experiments with non-toxic components (fibronectin or serum) produce a different QCM-D response than that observed for the toxic chemicals, suggesting the use of Df plot signatures for the possible differentiation between cytotoxic or non-cytotoxic effects. Observations obtained by QCM-D signals are confirmed by conducting fluorescence microscopy at the same time. Our results show that a fast (few minutes) sensing response can be obtained in situ and in real-time. The conclusions from this study suggest that QCM-D can potentially be used in biodetection for applications in drug screening tests and diagnosis.

摘要

石英晶体微天平与耗散监测(QCM-D)用于实时原位检测活原生内皮细胞对不同细胞形态试剂的细胞骨架变化;即表面活性剂 Triton-X 100(TX-100)和细菌脂多糖(LPS)。可重现的耗散与频率(Df)图谱提供了内皮细胞与细胞形态试剂相互作用的独特特征。虽然 TX-100 的 QCM-D 响应可以分为两个步骤(观察到预期的细胞裂解之前介质渗透压的变化),但 LPS 导致不同的单相信号。通过分析获得的 Df 图谱,进行补充分析以评估 TX-100 和 LPS 通过 BAEC 应激的 QCM-D 响应的可能竞争效应。与毒性化学物质相比,非毒性成分(纤连蛋白或血清)的实验产生了不同的 QCM-D 响应,这表明可以使用 Df 图谱特征来区分细胞毒性或非细胞毒性作用。通过 QCM-D 信号获得的观察结果同时通过荧光显微镜进行确认。我们的结果表明,可以原位实时获得快速(几分钟)的传感响应。这项研究的结论表明,QCM-D 有可能用于生物检测,用于药物筛选测试和诊断。

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