Centre for Biomedical Science Research, School of Clinical and Applied Sciences, Leeds Beckett University, Leeds LS1 3HE, United Kingdom.
Centre for Biomedical Science Research, School of Clinical and Applied Sciences, Leeds Beckett University, Leeds LS1 3HE, United Kingdom.
Biosens Bioelectron. 2018 Aug 15;113:46-51. doi: 10.1016/j.bios.2018.04.052. Epub 2018 Apr 26.
The measurement of pH is important throughout many biological systems, but there are limited available technologies to enable its periodical monitoring in the complex, small volume, media often used in cell culture experiments across a range of disciplines. Herein, pad printed electrodes are developed and characterised through modification with: a commercially available fullerene multiwall carbon nanotube composite applied in Nafion, casting of hydrophobic ubiquinone as a pH probe to provide the electrochemical signal, and coated in Polyethylene glycol to reduce fouling and potentially enhance biocompatibility, which together are proven to enable the determination of pH in cell culture media containing serum. The ubiquinone oxidation peak position (E) provided an indirect marker of pH across the applicable range of pH 6-9 (R = 0.9985, n = 15) in complete DMEM. The electrochemical behaviour of these sensors was also proven to be robust; retaining their ability to measure pH in cell culture media supplemented with serum up to 20% (v/v) [encompassing the range commonly employed in cell culture], cycled > 100 times in 10% serum containing media and maintain > 60% functionality after 5 day incubation in a 10% serum containing medium. Overall, this proof of concept research highlights the potential applicability of this, or similar, electrochemical approaches to enable to detection or monitoring of pH in complex cell culture media.
在许多生物系统中，pH 值的测量都很重要，但可用的技术有限，无法在跨学科的细胞培养实验中经常使用的复杂、小体积介质中定期监测 pH 值。在此，通过对印刷电极进行修饰，开发并表征了这种电极：一种商业上可用的富勒烯多壁碳纳米管复合材料应用于 Nafion 中，将疏水性泛醌铸造成 pH 探针以提供电化学信号，并涂覆聚乙二醇以减少污垢并可能提高生物相容性，事实证明，这三者结合在一起能够在含有血清的细胞培养基中测定 pH 值。在完整的 DMEM 中，泛醌氧化峰位置 (E) 在 pH 6-9 的适用范围内（R = 0.9985，n = 15）提供了 pH 的间接标志物。这些传感器的电化学行为也被证明是稳健的；它们仍然能够在含有高达 20%（v/v）血清的细胞培养基中测量 pH 值（涵盖细胞培养中常用的范围），在含有 10%血清的介质中循环 > 100 次，并在含有 10%血清的介质中孵育 5 天后保持 > 60%的功能。总的来说，这项概念验证研究强调了这种或类似电化学方法在复杂细胞培养基中检测或监测 pH 值的潜在适用性。
