In‐Sequence High‐Specificity Dual‐Reporter Unlocking of Fluorescent Probe Enables the Precise Identification of Atherosclerotic Plaques

The formation of atherosclerotic plaques is the root cause of various cardiovascular diseases (CVDs). Effective CVD interventions thus call for precise identification of the plaques to aid clinical assessment, diagnosis, and treatment of such diseases. In this study, we introduce a dual‐target sequentially activated fluorescence reporting system, termed in‐sequence high‐specificity dual‐reporter unlocking (iSHERLOCK), to precisely identify the atherosclerotic plaques in vivo and ex vivo. ISHERLOCK was achieved by creating a three‐in‐one fluorescent probe that permits highly specific and sensitive detection of lipid droplets and hypochlorous acid via “off‐on” and ratiometric readouts, respectively. Based on this format, the upregulated lipid accumulation and oxidative stress—the two hallmarks of atherosclerosis (AS)—were specifically measured in the atherosclerotic plaques, breaking through the barrier of precise tissue biopsy of AS and thus aiding effective CVD stewardship. An in‐sequence high‐specificity dual‐reporter unlocking platform, termed iSHERLOCK, is delicately designed to detect lipid droplets and HClO via fluorescent “off–on” and ratiometric readouts, respectively. Based on iSHERLOCK, the two hallmarks of atherosclerosis including upregulated lipid accumulation and oxidative stress were quantitatively measured in the plaques.