Enzymes play a critical role in most complex biochemical processes. Some of them can be regarded as biomarkers for disease diagnosis. Taking advantage of aggregation-induced emission (AIE)-based biosensors, a series of fluorogens with AIE characteristics (AIEgens) have been designed and synthesized for the detection and imaging of enzymes. In this work, we summarized the advances in AIEgens-based probes and sensing platforms for the fluorescent detection of enzymes, including proteases, phosphatases, glycosidases, cholinesterases, telomerase and others. The AIEgens involve organic dyes and metal nanoclusters. This work provides valuable references for the design of novel AIE-based sensing platforms.
Taking advantage of high porosity, large surface area, tunable nanostructures and ease of functionalization, metal-organic frameworks (MOFs) have been popularly applied in different fields, including adsorption and separation, heterogeneous catalysis, drug delivery, light harvesting, and chemical/biological sensing. The abundant active sites for specific recognition and adjustable optical and electrical characteristics allow for the design of various sensing platforms with MOFs as promising candidates. In this review, we systematically introduce the recent advancements of MOFs-based fluorescent chemosensors and biosensors, mainly focusing on the sensing mechanisms and analytes, including inorganic ions, small organic molecules and biomarkers (e.g., small biomolecules, nucleic acids, proteins, enzymes, and tumor cells). This review may provide valuable references for the development of novel MOFs-based sensing platforms to meet the requirements of environment monitoring and clinical diagnosis.
Fluorescence-assisted digital counting analysis allowed sensitive quantification of targets by measuring individual fluorescent labels. However, traditional fluorescent labels suffered from low brightness, small size, and sophisticated preparation procedures. Herein, engineering fluorescent dye-stained cancer cells with magnetic nanoparticles were proposed to construct singlecell probes for fluorescence-assisted digital counting analysis by quantifying the target-dependent binding or cleaving events. Various engineering strategies of cancer cells including biological recognition and chemical modification were developed for rationally designing single-cell probes. Introduction of suitable recognition elements into single-cell probes allowed digital quantification of each target-dependent event via counting the colored single-cell probes in the representative image taken using a confocal microscope. The reliability of the proposed digital counting strategy was corroborated by traditional optical microscopy-and flow cytometry-dependent counting technologies. The advantages of single-cell probes, including high brightness, big size, ease of preparation, and magnetic separation, contributed to the sensitive and selective analysis of targets of interest. As proof-to-concept assays, indirect analysis of exonuclease III (Exo III) activity, as well as direct quantitation of cancer cells, were investigated, and the potential in biological sample analysis was also assessed. This sensing strategy will open a new avenue for the development of biosensors.
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