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Ratiometric fluorescent pH-sensitive polymers for high-throughput monitoring of extracellular pH

机译：用于细胞外pH的高通量监测的比率荧光pH敏感聚合物

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Extracellular pH has a strong effect on cell metabolism and growth. Precisely detecting extracellular pH with high throughput is critical for cell metabolism research and fermentation applications. In this research, a series of ratiometric fluorescent pH sensitive polymers are developed and the ps-pH-neutral is characterized as the best one for exclusive detection of extracellular pH. Poly( N -(2-hydroxypropyl)methacrylamide) (PHPMA) is used as the host polymer to increase the water solubility of the pH sensitive polymer without introducing cell toxicity. The fluorescent emission spectra from the polymeric sensor under excitation at the isosbestic point 455 nm possess two fluorescence peaks at 475 nm and 505 nm, which have different responding trends to pH. This enables the polymer to detect pH using fluorescent maxima at 475 nm and 505 nm ( I _(475 nm) / I _(505 nm) ) ratiometrically. The cell impermeability ensures the sensor can solely detect the environmental pH. The sensor is tested to detect the extracellular pH of bacteria or eukaryotic cells in high throughput assays using a microplate reader. Results showed that the pH sensor can be used for high throughput detection of extracellular pH with high repeatability and low photobleaching effect.
机译：细胞外pH对细胞代谢和生长具有很强的影响。精确地检测高通量的细胞外pH对于细胞代谢研究和发酵应用至关重要。在该研究中，开发了一系列比例荧光pH敏感聚合物，并且PS-pH中性的特征是用于专用检测细胞外pH的最佳选择。聚（N - （2-羟丙基）甲基丙烯酰胺）（pHPMA）用作宿主聚合物，以增加pH敏感聚合物的水溶性而不引入细胞毒性。来自Isosbeste 455nm的激发的聚合物传感器的荧光发射光谱具有在475nm和505nm处的两个荧光峰，其具有不同的响应趋势至pH。这使得聚合物能够在475nm和505nm（I _（475nm）/ I _（505nm））下使用荧光最小值来检测pH值。细胞不透毛可确保传感器可以单独检测环境pH。使用微孔板读卡器测试传感器以检测高通量测定中的细菌或真核细胞的细胞外pH。结果表明，PH传感器可用于具有高可重复性和低光漂白效果的细胞外pH的高通量检测。

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