Ferroptosis modulation represents a pioneering therapeutic approach for non-small-cell lung cancer (NSCLC), where precise monitoring and regulation of ferroptosis levels are pivotal for achieving optimal therapeutic outcomes. Cisplatin (Cis), a widely used chemotherapy drug for NSCLC, demonstrates remarkable therapeutic efficacy, potentially through its ability to induce ferroptosis and synergize with other treatments. However, in vivo studies of ferroptosis face challenges due to the scarcity of validated biomarkers and the absence of reliable tools for real-time visualization. Biothiols emerge as suitable biomarkers for ferroptosis, as their concentrations decrease significantly during this process. To address these challenges, fluorescence/photoacoustic (PA) bimodal imaging offers a promising solution by providing more accurate in vivo information on ferroptosis. Therefore, the development of methods to detect biothiols using fluorescence/PA bimodal imaging is highly desirable for visualizing ferroptosis in NSCLC. In this study, we designed and constructed two activatable near-infrared (NIR) fluorescent/PA bimodal imaging probes specifically for visualizing ferroptosis by monitoring the fluctuations in biothiol levels. These probes exhibited excellent bimodal response performance in solution, cells, and tumors. Furthermore, they were successfully applied for real-time monitoring of biothiol changes during the ferroptosis process in NSCLC cells and tumors. Importantly, our findings revealed that the combined use of erastin and cisplatin exacerbates the consumption of biothiols, suggesting an enhancement of ferroptosis in NSCLC. This work not only provides powerful tools for monitoring in vivo ferroptosis but also facilitates the study of ferroptosis mechanisms and holds the potential to further advance the treatment of NSCLC.