Firestone Institute for Respiratory Health-Division of Respirology, Department of Medicine, McMaster University, Hamilton, ON, L8N 4A6, Canada.
Department of Mechanical Engineering, McMaster University, Hamilton, ON, L8S 4L7, Canada.
Sci Rep. 2021 Feb 10;11(1):3477. doi: 10.1038/s41598-021-83189-x.
In many biological systems, pH can be used as a parameter to understand and study cell dynamics. However, measuring pH in live cell culture is limited by the sensor ion specificity, proximity to the cell surface, and scalability. Commercially available pH sensors are difficult to integrate into a small-scale cell culture system due to their size and are not cost-effective for disposable use. We made PHAIR-a new pH sensor that uses a micro-wire format to measure pH in vitro human airway cell culture. Tungsten micro-wires were used as the working electrodes, and silver micro-wires with a silver/silver chloride coating were used as a pseudo reference electrode. pH sensitivity, in a wide and narrow range, and stability of these sensors were tested in common standard buffer solutions as well as in culture media of human airway epithelial cells grown at the air-liquid interface in a 24 well cell culture plate. When measuring the pH of cells grown under basal and challenge conditions using PHAIR, cell viability and cytokine responses were not affected. Our results confirm that micro-wire-based sensors have the capacity for miniaturization and detection of diverse ions while maintaining sensitivity. This suggests the broad application of PHAIR in various biological experimental settings.
在许多生物系统中，pH 值可以作为一个参数来了解和研究细胞动力学。然而，在活细胞培养中测量 pH 值受到传感器离子特异性、接近细胞表面的程度和可扩展性的限制。由于尺寸原因，商业上可用的 pH 传感器难以集成到小规模细胞培养系统中，并且由于一次性使用，其成本效益不高。我们制作了 PHAIR，这是一种新型 pH 传感器，使用微线格式在体外人呼吸道细胞培养中测量 pH 值。钨微线被用作工作电极，而带有银/氯化银涂层的银微线被用作伪参比电极。在常见的标准缓冲溶液以及在 24 孔细胞培养板中在气液界面生长的人呼吸道上皮细胞的培养基中，测试了这些传感器在宽范围和窄范围以及稳定性方面的 pH 敏感性。当使用 PHAIR 测量在基础和挑战条件下生长的细胞的 pH 值时，细胞活力和细胞因子反应不受影响。我们的结果证实，基于微线的传感器具有小型化和检测多种离子的能力，同时保持灵敏度。这表明 PHAIR 在各种生物实验设置中有广泛的应用。
