Dual-Targeted Fluorogenic Probes for Precise In Vivo Imaging: Design Principles, Activation Mechanisms, and Applications

Abstract

Fluorogenic probes with "off-on" fluorescence signals have emerged as powerful tools for biosensing and bioimaging of biomolecules in living systems. Conventional single-target probes, however, often suffer from false-positive signals due to non-specific activation in non-target tissues or the diffusion of activated fluorescent products. To address these limitations, dual-targeted fluorogenic probes (DTFPs) have been developed, which simultaneously target two biomarkers to enhance detection specificity and minimize false-positive outcomes. DTFPs are designed to activate or retain fluorescence only when both biomarkers are present within a targeted region, enabling precise in vivo imaging of pathological conditions such as tumors and inflammation. This review highlights recent advances in DTFP development, focusing on their design principles, activation mechanisms, and applications in biosensing and bioimaging. We also discuss current challenges and future directions for DTFP research, aiming to inspire the design of next-generation probes with improved accuracy and specificity. By providing a comprehensive overview of DTFPs, this review seeks to advance their potential for transformative applications in biomedical imaging and diagnostics.

Keywords

Fluorescence imaging / In Vivo / Dual-targeted / Fluorogenic probes / Biomarkers / Bioimaging

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