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原位阻抗生物传感器芯片监测细菌生物膜形成过程。

Monitoring of bacteria biofilms forming process by in-situ impedimetric biosensor chip.

机构信息

School of Chemistry and Chemical Engineering, Chongqing University, Chongqing 400030, China; Key Disciplines Lab of Novel Micro-Nano Devices and System Technology, Chongqing University, Chongqing 400030, China; National Center for International Research of Micro/Nano-System and New Material Technology, Chongqing University, Chongqing 400030, China.

School of Chemistry and Chemical Engineering, Chongqing University, Chongqing 400030, China; Key Disciplines Lab of Novel Micro-Nano Devices and System Technology, Chongqing University, Chongqing 400030, China; National Center for International Research of Micro/Nano-System and New Material Technology, Chongqing University, Chongqing 400030, China; School of Optoelectronics Engineering, Chongqing University, Chongqing 400044, China.

出版信息

Biosens Bioelectron. 2018 Jul 30;112:86-92. doi: 10.1016/j.bios.2018.04.019. Epub 2018 Apr 12.

Abstract

A biosensor chip integrated interdigital microelectrodes was proposed and applied to monitor the formation process of Salmonella and E. coli biofilms in this paper. The biosensor chip was composed of a glass substrate with interdigital microelectrodes and PDMS layer with micro cavities. The electrochemical impedance spectroscopy (EIS) of Salmonella and E. coli biofilms was measured by the biosensor chip using alternating voltage of 100 mv in the frequency range from 1 Hz to 100 kHz for 48 h. It was illustrated that the changes of impedance spectroscopy of biofilms occurred with culture time. Furthermore, impedance spectroscopy of biofilms was fitted by an equivalent circuit model including the biofilms capacitance (C) and the biofilms resistance (R) parameters. The results indicated that the C presented a tendency to decrease first and then rise with culture time, while the R was in the opposite direction. These changing trends were consistent with the formation process of biofilms that bacteria adhered to electrodes surface, and then formed mature biofilms, finally escaped from biofilms. In addition, it was also demonstrated that the changing trends of C and R with culture time were quite different between Salmonella and E. coli. The results obtained by impedance detection were in accordance with the results of using crystal violet staining to analyze biofilms formation process, under the same conditions for bacterial culture. The biosensor chip provided a promising platform for further study of biofilms owing to its unique advantages of real time, continuity, and non-invasion for bacteria biofilms detection and in-situ monitoring.

摘要

本文提出并应用了一种集成叉指微电极的生物传感器芯片,用于监测沙门氏菌和大肠杆菌生物膜的形成过程。该生物传感器芯片由带有叉指微电极的玻璃基底和带有微腔的 PDMS 层组成。使用生物传感器芯片,在 1 Hz 至 100 kHz 的频率范围内,以 100 mV 的交流电压对沙门氏菌和大肠杆菌生物膜进行了 48 小时的电化学阻抗谱(EIS)测量。结果表明,生物膜的阻抗谱随培养时间而变化。此外,通过包含生物膜电容(C)和生物膜电阻(R)参数的等效电路模型对生物膜的阻抗谱进行拟合。结果表明,C 随培养时间呈现先减小后增大的趋势,而 R 则相反。这些变化趋势与生物膜的形成过程一致,即细菌首先附着在电极表面,然后形成成熟的生物膜,最后从生物膜中逃脱。此外,还表明 C 和 R 随培养时间的变化趋势在沙门氏菌和大肠杆菌之间有很大的不同。在相同的细菌培养条件下,阻抗检测的结果与使用结晶紫染色分析生物膜形成过程的结果一致。由于该生物传感器芯片具有实时、连续、非侵入式检测细菌生物膜的独特优势,因此为生物膜的进一步研究提供了一个有前途的平台。

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