Real-Time Tracking of Mitochondrial O2•– and Polarity Fluctuations in Ferroptosis via a Polysiloxane-Supported Dual-Channel Fluorescent Probe

Abstract Ferroptosis is intricately linked to mitochondrial dysfunction, particularly the abnormal accumulation of superoxide anions (O2•–) and alterations in membrane polarity. To decipher the spatiotemporal relationship between these factors, we report the construction of Si-Fur-Hem, a polysiloxane-based fluorescent probe designed for dual-responsive imaging. By grafting an ICT-based polarity sensor and a triggered hemicyanine onto a flexible polysiloxane chain, the probe achieves simultaneous monitoring of the mitochondrial microenvironment. The intrinsic positive charge of the hemicyanine unit facilitates specific mitochondrial localization. Experimental results demonstrate that Si-Fur-Hem can dynamically track the synergism between production and polarity shifts during ferroptosis. This bifunctional imaging platform not only clarifies the molecular mechanisms of ferroptosis but also serves as a robust tool for screening therapeutic strategies targeting mitochondrial homeostasis.