Optochemical sensors are actively used in cell analysis, however existing systems have limitations with respect to their robustness and analytical performance. We have developed advanced multimodal and multi-parametric solid-state pH sensors for cell analysis based on hydrophobic protonable metal-free porphyrins, such as octaethylporphine (OEP), and octaethylporphine-ketone (OEPK), as fluorescent pH indicators. The internally referenced ratiometric intensity and nanosecond lifetime-based versions of the pH sensors were developed and also multiplexed with the O₂ sensors based on phosphorescent PtOEP dye. We optimised the key parameters of the pH sensor, including: dye encapsulation matrix, the type and concentration of proton transfer reagent, measurement pH range and pKa, dye concentrations and cross-talk with the O₂ sensor. Subsequently, sensor coatings were deposited on common substrates used in cell analysis (96-well plates), fine-tuned for their operational performance, dual O₂/pH sensing functionally and the ability to measure Extracellular Acidification (ECAR) and Oxygen Consumption (OCR) rates in biological samples containing cells. The optimised sensors with stable internally referenced calibrations, convenient spectral characteristics and low cytotoxicity, were demonstrated with cultured cells and 3D spheroid structures, measuring their ECAR, OCR and responses to stimulation. These pH and dual pH/O₂ sensors are well-suited for detailed metabolic studies of biological samples on widely available laboratory equipment.

光化学传感器在细胞分析中得到了广泛应用，但现有系统在其稳健性和分析性能方面存在局限性。我们开发了先进的多模式和多参数固态 pH 传感器，用于基于疏水质子无金属卟啉（如八乙基卟啉 (OEP) 和八乙基卟啉酮 (OEPK) 作为荧光 pH 指示剂）的细胞分析。开发了内部参考的比率强度和基于纳秒寿命的 pH 传感器版本，并与 O _2复用基于磷光 PtOEP 染料的传感器。_{我们优化了pH传感器的关键参数，包括：染料封装基质、质子转移试剂的类型和浓度、测量pH范围和pKa、染料浓度和与O 2}传感器的串扰。随后，传感器涂层被沉积在细胞分析中使用的普通基底（96 孔板）上，针对其操作性能进行了微调，双 O ₂/pH 传感功能和测量含有细胞的生物样品中的细胞外酸化 (ECAR) 和耗氧 (OCR) 率的能力。优化的传感器具有稳定的内部参考校准、方便的光谱特性和低细胞毒性，并通过培养细胞和 3D 球体结构进行了验证，测量了它们的 ECAR、OCR 和对刺激的响应。这些 pH 和双 pH/O ₂传感器非常适合在广泛可用的实验室设备上对生物样品进行详细的代谢研究。