ReviewSensors (Basel, Switzerland)2026
Fluorescent Probes for Aldehyde Detection in Biological Systems: Design Principles, Spectroscopy, and Emerging Applications.
Review in Sensors (Basel, Switzerland), 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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2 authors.
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Abstract
Reactive aldehydes-formaldehyde (FA), malondialdehyde (MDA), 4-hydroxynonenal (4-HNE), and acrolein-occupy a central role in epigenetic regulation, lipid peroxidation, ferroptosis, and cardiovascular disease, yet their transient nature and low intracellular concentrations have long made them difficult to quantify in living systems. Over the past decade, reaction-based small-molecule fluorescent probes have emerged as the principal tool for addressing this challenge, and this review provides a systematic account of that progress. We compare the six main conjugation chemistries that underlie current probe design-2-aza-Cope/Mannich cascade, hydrazone/oxime formation, Michael addition, Schiff base condensation, and 2-aminothiophenol cyclization-alongside the three photophysical strategies used to convert these reactions into quantitative signals: intensity turn-on, ratiometric dual-emission, and fluorescence lifetime imaging (FLIM). Attention is given to advances reported between 2020 and 2025, including organelle-targeted ratiometric formaldehyde sensors (MitoRFAP-2, NucRFAP-2), the first ratiometric acrolein probe for visualizing ferroptosis, lysosome-targeted malondialdehyde reporters for atherosclerosis staging, and near-infrared-compatible platforms validated in three-dimensional organoids and in vivo models. This review also critically examines the limitations that continue to constrain the field, including insufficient selectivity testing under physiologically relevant conditions, the pH-dependence of hydrazone equilibria, the frequently overlooked distinction between probes that report free aldehyde concentration and those that report ALDH enzyme activity, and the continued absence of reversible, real-time sensors. Closing these gaps will determine whether aldehyde imaging grows from a set of smart probes into a quantitative, widely trusted platform for studying redox and carbonyl biology in living systems. Reaching that point will depend on three criteria: better NIR fluorophores, effective bioconjugation chemistry, and machine-learning tools that guide probe design.
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Registered trials
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