A Mitochondrial Microenvironment-Responsive Fluorescent Sensor Array for Intelligent Analysis of Lung Cancer Cells and Chemotherapeutic Efficacy

Abstract Accurate discrimination of lung cancer subtypes and evaluation of chemotherapeutic efficacy remain critical challenges in precision oncology owing to the heterogeneity of tumor cellular microenvironments. Herein, we report a mitochondrial microenvironment-responsive fluorescence sensor array, MR-SA, for intelligent analysis of lung cancer cells and chemotherapeutic efficacy. Four mitochondria-targeting fluorescent probes were rationally designed by integrating indolium salts with environment-sensitive fluorophores, enabling selective sensing of mitochondrial hypoxia, polarity, pH, and viscosity in living cells. Benefiting from distinct cross-reactive fluorescence response patterns and broad emission coverage ranging from 573 to 734 nm, MR-SA generated multidimensional fluorescence fingerprints for different lung cancer subtypes while minimizing spectral crosstalk. The chemotherapy-induced mitochondrial alterations were monitored using MR-SA and combined with outlier analysis for evaluation of chemotherapeutic efficacy and drug resistance in nonsmall cell lung cancer cells. Furthermore, MR-SA fluorescence images were integrated with ResNet, a deep-learning framework, enabling rapid and accurate blind classification of lung cancer cells with a recognition accuracy of 99.71%. Thus, MR-SA successfully distinguished lung adenocarcinoma, lung squamous cell carcinoma, large cell carcinoma, and small cell lung cancer cells through differential mitochondrial microenvironment profiling. This work establishes a versatile fluorescence sensing strategy for multidimensional mitochondrial analysis and provides a promising analytical platform for intelligent cancer cell profiling and chemotherapeutic evaluation.