HOCl and viscosity dual-responsive fluorescent probe for accurate discrimination between early hepatocellular carcinoma and acute liver injury.

The accurate differentiation between early-stage hepatocellular carcinoma (HCC) and acute liver injury (ALI) remains a critical yet unresolved challenge in clinical practice, as these conditions share overlapping symptoms and biomarkers, often leading to misdiagnosis with conventional techniques. Although fluorescent probes offer potential for high-sensitivity imaging, existing designs typically detect only a single parameter and lack subcellular targeting precision. Consequently, a methodology that simultaneously visualizes multiple pathology-specific biomarkers within a key organelle for precise discrimination is urgently needed. Here, we present a lipid droplet-targeted, dual-channel fluorescent probe, TPA-DCN-TPE, which independently monitors microenvironmental viscosity and hypochlorous acid (HOCl). This probe exhibits a remarkable 1532-fold fluorescence enhancement in response to viscosity (red channel) and a 363-fold turn-on response to HOCl (green channel). Crucially, we discovered a distinct red-to-green signal ratio that clearly discriminates HCC (high ratio) from ALI (low ratio) in vivo, ex vivo, and in tissue sections. Our work establishes a quantifiable optical criterion for differentiating these liver pathologies, thereby addressing a major diagnostic gap. We anticipate this dual-parameter imaging strategy will advance the precision diagnosis of liver diseases and provide a versatile platform for studying organelle-specific microenvironments in other metabolic and inflammatory disorders.