Institute for Biocomplexity and Informatics, University of Calgary, 2500 University Driver NW, Calgary, Alberta, Canada.
Biosens Bioelectron. 2010 Mar 15;25(7):1774-80. doi: 10.1016/j.bios.2009.12.025. Epub 2009 Dec 29.
This paper presents a simple mathematical model to predict the impedance data acquired by electric cell-substrate impedance sensing (ECIS) at frequencies between 25 Hz and 60 kHz. With this model, the equivalent resistance (R) and capacitance (C) of biological samples adhered on gold surfaces could be more precisely measured at 4 kHz. ECIS applications were extended for real-time monitoring of living bacteria cultivated in Luria Bertani (LB) culture medium by two different approaches. In the former, we used a ferri/ferrocyanide redox couple in LB medium as an indicator for bacterial multiplication. Because the redox couple was toxic to some bacteria, we developed a second approach, in which l-cysteine self-assembled monolayers (SAM) on gold electrodes were used to detect living bacteria. The l-cysteine SAM could also be detected by ECIS. Unlike traditional impedance microbiological methods which need special culture media with low ions, our procedures significantly enhanced signal/noise ratios so bacteria could be detected in general purpose culture media. It was easy and convenient to obtain bacterial doubling times and evaluate the resistance of bacteria to antibiotics from ECIS spectra.
本文提出了一个简单的数学模型,用于预测在 25 Hz 至 60 kHz 频率下通过电细胞-基底阻抗感应 (ECIS) 获得的阻抗数据。使用该模型,可以更精确地在 4 kHz 测量附着在金表面上的生物样品的等效电阻 (R) 和电容 (C)。ECIS 应用通过两种不同的方法扩展到实时监测在 LB 培养基中培养的活细菌。在前者中,我们使用 LB 培养基中的铁/亚铁氰化物氧化还原偶联物作为细菌繁殖的指示剂。由于氧化还原偶联物对某些细菌有毒,我们开发了第二种方法,其中金电极上的 l-半胱氨酸自组装单层 (SAM) 用于检测活细菌。l-半胱氨酸 SAM 也可以通过 ECIS 检测到。与需要低离子特殊培养基的传统阻抗微生物方法不同,我们的方法大大提高了信号/噪声比,因此可以在普通培养基中检测到细菌。从 ECIS 谱中很容易获得细菌的倍增时间并评估细菌对抗生素的耐药性。