Theranostic Fluorescent Probes.

The ability to gain spatiotemporal information, and in some cases achieve spatiotemporal control, in the context of drug delivery makes theranostic fluorescent probes an attractive and intensely investigated research topic. This interest is reflected in the steep rise in publications on the topic that have appeared over the past decade. Theranostic fluorescent probes, in their various incarnations, generally comprise a fluorophore linked to a masked drug, in which the drug is released as the result of certain stimuli, with both intrinsic and extrinsic stimuli being reported. This release is then signaled by the emergence of a fluorescent signal. Importantly, the use of appropriate fluorophores has enabled not only this emerging fluorescence as a spatiotemporal marker for drug delivery but also has provided modalities useful in photodynamic, photothermal, and sonodynamic therapeutic applications. In this review we highlight recent work on theranostic fluorescent probes with a particular focus on probes that are activated in tumor microenvironments. We also summarize efforts to develop probes for other applications, such as neurodegenerative diseases and antibacterials. This review celebrates the diversity of designs reported to date, from discrete small-molecule systems to nanomaterials. Our aim is to provide insights into the potential clinical impact of this still-emerging research direction.

De Novo Design of a Highly Stable Ratiometric Probe for Long-Term Continuous Imaging of Endogenous HClO Burst.

Long-term continuous imaging of endogenous HClO burst is of great importance for the elucidation of various physiological or pathological processes. However, most of the currently reported HClO probes have failed to achieve this goal due to their insufficient photobleaching resistance under a laser source. Herein, a highly stable ratiometric probe, HFTC-HClO 1, which is capable of continuously monitoring endogenous HClO burst over a long period of time, has been judiciously developed. Briefly, the de novo development of HFTC-HClO 1 mainly involved three main steps: (1) novel coumarins (HFTC 1-5) were designed and synthesized; (2) the most stable scaffold, HFTC 3, was selected through dye screening and cell imaging validation; and (3) based on HFTC 3, three candidate HClO probes were constructed, and HFTC-HClO 1 was finally selected due to its superior sensing properties toward HClO. Furthermore, HFTC-HClO 1 can quantitatively measure HClO levels in various real samples with excellent recovery (>90.4%), and the use of HFTC-HClO 1-coated test strips for qualitative analysis of HClO in real samples was also achieved. In addition, the application of HFTC-HClO 1 for long-term continuous monitoring of intracellular HClO burst was successfully demonstrated. Significantly, HFTC-HClO 1 was able to visualize HClO generated in the rheumatoid arthritis mouse model.

Engineered Collaborative Size Regulation and Shape Engineering of Tremella-Like Au-MnOx for Highly Sensitive Bimodal-Type Lateral Flow Immunoassays.

Engineered collaborative size regulation and shape engineering of multi-functional nanomaterials (NPs) offer extraordinary opportunities for improving the analysis performance. It is anticipated to address the difficulty in distinguishing color changes caused by subtle variations in target concentrations, thereby facilitating the highly sensitive analysis of lateral flow immunoassays (LFIAs). Herein, tremella-like gold-manganese oxide (Au-MnOx ) nanoparticles with precise MnCl2 regulation are synthesized as immuno signal tracers via a facile one-step redox reaction in alkaline condition at ambient temperature. Avail of the tunable elemental composition and anisotropy in morphology, black-colored tremella-like Au-MnOx exhibits superb colorimetric signal brightness, enhanced antibody coupling efficiency, marvelous photothermal performance, and unrestricted immunological recognition affinity, all of which facilitate highly sensitive multi-signal transduction patterns. In conjunction with the handheld thermal reader device, a bimodal-type LFIA that combines size-regulation- and shape-engineering-mediated colorimetric-photothermal dual-response assay (coined as the SSCPD assay) with a limit of detection of 0.012 ng mL-1 for ractopamine (RAC) monitoring is achieved by integrating Au-MnOx with the competitive-type immunoreaction. This work illustrates the effectiveness of this strategy for establishing high-performance sensing, and the SSCPD assay may be extended to a wide spectrum of future point-of-care (POC) diagnostic applications.

Real-Time Fluorescence Screening Platform for Meat Freshness.

Meat's freshness is closely related to food safety and human health and has received increasing attention nowadays. To on-site visually screen meat freshness in a fast and non-destructive manner, we rationally constructed a series of fluorescent probes (JDCN, JDNS, and JDPY) with distinct electron-withdrawing substitution groups based on julolidine-fused coumarin. These probes underwent an aza-Michael addition followed by an elimination reaction with cadaverine to generate a colorimetric and ratiometric fluorescence response, and their sensing performance was rationally enhanced by improving the electron-withdrawing strength of substitution groups. Particularly, JDCN with a dicyanovinyl group as the reaction site exhibited outstanding sensing performance including rapid response (∼60 s), high selectivity, and low detection limit (14 nM). Furthermore, JDCN was fabricated into test kits to detect cadaverine vapor with a high-contrast fluorescence change from red to green. Based on two-color visualization of cadaverine vapor, on-site non-contact and non-destructive monitoring of meat freshness was successfully achieved. The good sensing performance rendered JDCN test kits a promising real-time fluorescence screening platform for rapid, non-destructive, and accurate evaluation of meat freshness.

A Single Fluorescent Chemosensor for Simultaneous Discriminative Detection of Gaseous Phosgene and a Nerve Agent Mimic.

A fluorescent chemosensor has been developed for discriminative detection of phosgene and a nerve agent mimic diethyl chlorophosphate (DCP), which was comprised of an anthracene-carboxyimide fluorophore and o-phenylenediamine (OPD) reaction site. Upon phosphorylation of OPD, the chemosensor displays an obvious fluorescence turn-on response toward DPC at 588 nm with instant response and a low detection limit (88 nM). By contrast, the chemosensor exhibits a colorimetric and fluorescence enhancement response at 500 nm toward phosgene with fast response (<2 min), high selectivity, and a low detection limit (72 nM). Furthermore, chemosensor-loaded test membrane was fabricated for real-time, portable and efficient discriminative detection of trace amounts of gaseous phosgene and DCP vapor with different optical responses.

In Vivo Imaging of Endogenously Produced HClO in Zebrafish and Mice Using a Bright, Photostable Ratiometric Fluorescent Probe.

A high brightness red fluorescent probe (S-BODIPY) has been developed for the sensitive and specific imaging of HClO/ClO- in vitro and in vivo. This probe exhibits some distinctive features such as excellent resistance to photobleaching, a high fluorescence brightness, high selectivity, as well as a good biocompatibility. Upon oxidation of the thio-ether group into sulfoxide, the probe showed a noticeable ratiometric fluorescence response toward ClO- with fast response (within 30 s) and a low detection limit (59 nM). The probe demonstrated the successful imaging of exogenous and endogenous HClO/ClO- in living HeLa cells, zebrafish, and mice with high signal-to-noise ratios. S-BODIPY allows for the real-time monitoring the level of ClO- in living cells by ratiometric fluorescence imaging, opening up exciting prospects to develop red and even near-infrared BODIPYs with high brightness and good photostability for in vivo imaging.

Colorimetric and Ratiometric Chemosensor for Visual Detection of Gaseous Phosgene Based on Anthracene Carboxyimide Membrane.

In this work, we reported an anthracene carboxyimide-based chemosensor (AC-Phos) for colorimetric and ratiometric fluorescence detection of highly toxic phosgene, which displayed rapid response (<5 min) toward phosgene with a high selectivity and a low detection limit (2.3 nM). Furthermore, a facile testing membrane with a polystyrene immobilizing chemosensor has been fabricated for real-time visualizing of gaseous phosgene.

Regulating glutathione-responsiveness of naphthalimide-based fluorescent probes by an oxidation strategy.

Two naphthalimide-based fluorescent probes containing a thiomorpholine (Np-NS) or a sulfoxide-morpholine (Np-NSO) component are reported. The morpholine unit of non-fluorescent Np-NS and Np-NSO can transform into sulphone-morpholine and be accompanied by blue fluorescence upon oxidative stress, ascribed to the formation of sulphone-morpholine on probes. This sensing behavior displays that they can selectively respond to glutathione to generate a green emission by a sulfonamide-based detection moiety both in vitro and in living cells. Interestingly, the different oxidation states of a sulphur atom on a thiomorpholine ring can be utilized to regulate responsiveness of these probes towards glutathione. Such an oxidation strategy would provide a possibility for enhancing the response rate.

meso-Ester and carboxylic acid substituted BODIPYs with far-red and near-infrared emission for bioimaging applications.

A series of meso-ester-substituted BODIPY derivatives 1-6 are synthesized and characterized. In particular, dyes functionalized with oligo(ethylene glycol) ether styryl or naphthalene vinylene groups at the α positions of the BODIPY core (3-6) become partially soluble in water, and their absorptions and emissions are located in the far-red or near-infrared region. Three synthetic approaches are attempted to access the meso-carboxylic acid (COOH)-substituted BODIPYs 7 and 8 from the meso-ester-substituted BODIPYs. Two feasible synthetic routes are developed successfully, including one short route with only three steps. The meso-COOH-substituted BODIPY 7 is completely soluble in pure water, and its fluorescence maximum reaches around 650 nm with a fluorescence quantum yield of up to 15 %. Time-dependent density functional theory calculations are conducted to understand the structure-optical properties relationship, and it is revealed that the Stokes shift is dependent mainly on the geometric change from the ground state to the first excited singlet state. Furthermore, cell staining tests demonstrate that the meso-ester-substituted BODIPYs (1 and 3-6) and one of the meso-COOH-substituted BODIPYs (8) are very membrane-permeable. These features make these meso-ester- and meso-COOH-substituted BODIPY dyes attractive for bioimaging and biolabeling applications in living cells.

Synthesis and spectroscopic properties of some novel BODIPY dyes.

BODIPY dyes have some unique properties including high fluorescence quantum yield, large extinction coefficiency, narrow absorption and emission band. However, most of BODIPY dyes display short emission wavelength and small Stokes shift, which limits their applications in biosensing and bioimaging in vivo. For bioimaging application, a fluorescent dye with long emission wavelength and large Stokes shift is highly desired. To push the absorption and emission spectrum of BODIPY to red and even far-red region, a COOEt group was introduced to the meso position, and some aromatic group was attached to the 3, 5 position of BODIPY core. The structure of resulting compounds were comfirmed by 1H NMR, 13C NMR and HR-MS. Dye-1 displays a strong UV-Vis absorption band centered at 536 nm and a sharp emission band is located at 592 nm, which is significantly red-shifted (80 nm) compared to ordinary BODIPY analogs. In addition, the meso-COOEt substituted BODIPYs exhibit high quantum yield and red to far-red emission. Notably surprisingly, the meso-COOEt substituted BODIPYs display almost separated UV-Vis absorption and emission spectra with a large Stokes shift (-60 nm). Time-dependent density functional theory calculations were conducted to understand the structure-optical properties relationship, and it was revealed that the large Stokes shift was resulted from the geometric change from the ground state to the first excited singlet state. The spectroscopic properties of these BODIPY dyes display very subtle solvent-dependence effect. Furthermore, BODIPY was tested for its ability of imaging in living cells. The results indicate that Dye-1 is a water-soluble and membrane-permeable probe. Therefore, these BODIPYs are a new family dyes with excellent spectroscopic properties and can be good candidates for bioimaging in living cells.

Intramolecular cascade reaction sensing platform for rapid, specific and ultrasensitive detection of nitrite.

Nitrite is a carcinogenic substance in food. Excessive consumption of nitrite severely endangers human health. However, rapid and accurate quantification of nitrite by a simple tool is still very challenging. In this work, we designed a practical sensing platform based on 8-(o-phenylenediamine)-boron dipyrromethene (BDP-OPD) to determine nitrite in food. BDP-OPD can take a specific diazotization-cyclization cascade reaction with nitrite to form boron dipyrromethene (BODIPY), giving rise to a remarkable chromogenic reaction along with high contrast fluorescence turn-on response towards nitrite. BDP-OPD has high sensitivity, rapid response, and good selectivity. Furthermore, a portable smartphone-based fluorescence device integrated with a self-programmed Python program was fabricated, which has been successfully used to determine nitrite in food with the advantages of rapid response, low cost, ease of operation, portability, and satisfactory recoveries (92-112%). The good sensing performance rendered BDP-OPD a promising fluorescence platform for on-site visual detection of nitrite.

A practical chromogenic and fluorogenic dual-mode sensing platform for rapid quantification of sulfite in food.

Sulfur dioxide (SO2) and its derivatives (HSO3- and SO32-) are widely used in food-processing. Whereas excessive consumption of sulfur dioxide and its derivatives (>0.70 mg·kg-1day-1) severely endangers human health. In this work, we rationally constructed a practical dual-mode probe (dicyanomethylene)-1-methyl-1,4-dihydroquinolin-2-yl)vinyl)-1-methylquinolinium (QMN), which underwent a specific 1, 4-Michael addition with sulfite to afford a noticeable color change from pale yellow to red along with a high-contrast fluorescence turn-on response at 598 nm. QMN has the advantages of rapid response, high signal-to-noise ratio, excellent selectivity, good water-solubility, large Stokes shift and low detection limit (LOD = 31.9 nM). QMN has been successfully used to on-site visually determine sulfite in a diversity of foods with satisfactory recoveries (91.33-111.33 %) and high accuracy (93.74-98.71 %). Furthermore, a portable smartphone-based fluorescence sensing platform was fabricated for on-site determination of sulfite in food with good performance.

Endogenous CO imaging in bacterial pneumonia with a NIR fluorescent probe.

Bacterial pneumonia is a serious respiratory illness that poses a great threat to human life. Rapid and precise diagnosis of bacterial pneumonia is crucial for symptomatic clinical treatment. Endogenous carbon monoxide (CO) is regarded as a significant indicator of bacterial pneumonia; herein, we developed a near-infrared (NIR) probe for fluorescence and photoacoustic (PA) dual-mode imaging of endogenous CO in bacterial pneumonia. NO2-BODIPY could rapidly and specifically react with CO to produce strong NIR fluorescence as well as ratiometric PA signals. NO2-BODIPY has outstanding features including fast response, fluorescence/PA dual mode signals, good specificity, and a low limit of detection (LOD = 20.3 nM), which enables it to image endogenous CO in cells and bacterial pneumonia mice with high sensitivity and high contrast ratio. In particular, NO2-BODIPY has two-photon excited (1340 nm, σ1 = 1671 GM) NIR fluorescence and has been utilized to image endogenous CO in bacterial pneumonia mice with deep tissue penetration. NO2-BODIPY has been demonstrated a good capability of fluorescence/PA dual-mode imaging of CO in bacterial pneumonia mice, providing a precise manner to diagnose bacterial pneumonia.

A novel and photostable pH probe for selectively staining nuclei in living cells.

A novel pH probe based on hemi-cyanine was synthesized, which displays extremely large Stokes shift (122 nm) and good photostability. The probe responds to basic pH (pKa 8.32) with a colorimetric and fluorescence turn-on signal. Interestingly, the probe has good cell membrane permeability and could selectively stain nuclei in living cells.

A colorimetric and fluorescent sensor for non-destructive screening of the freshness of shrimp and fish.

The freshness of fish and shrimp is closely associated with food safety, hence it is a wide concern to develop a facile and effective method for fast, non-destructive and visual screening the freshness of fish and shrimp. Herein, we developed a chromogenic and fluorogenic sensor (RFCC) based on resorufin for sensing of biogenic amines including cadaverine and putrescine. RFCC underwent aminolysis with cadaverine or putrescine, displaying a remarkable fluorescence turn on response at 593 nm along with obvious color change from colorless to pink. RFCC was fabricated into test strips to sense cadaverine vapor, and the RGB value of test strips showed a good linear relationship with the concentration of cadaverine (0.5 - 8.2 × 103 ppm). The RFCC tag was used to in situ screen the freshness of fish and shrimp according to obvious fluorescence change, and satisfactory results were achieved. Furthermore, this test strip was validated by total volatile base nitrogen (TVBN), providing a simple, low cost and portable tool to screen the freshness of fish and shrimp for consumers and suppliers without expensive instrumentation.

Molecular engineering of fluorescence probe for real-time non-destructive visual screening of meat freshness.

Spoiled meat poses a great challenge to food security and human health, which should be addressed by the early monitoring and warning of the meat freshness. We herein exploited a molecular engineering strategy to construct a set of fluorescence probes (PTPY, PTAC, and PTCN) with phenothiazine as fluorophore and cyanovinyl as recognition site for the facile and efficient monitoring of meat freshness. These probes produce an obvious fluorescence color transition from dark red to bright cyan in response to cadaverine (Cad) through the nucleophilic addition/elimination reaction. The sensing performances were elaborately improved to achieve quick response (16 s), low detection limit (LOD = 3.9 nM), and high contrast fluorescence color change by enhancing the electron-withdrawing strength of cyanovinyl moiety. Furthermore, PTCN test strips were fabricated for portable and naked-eye detection of Cad vapor with fluorescence color change from crimson to cyan, and accurate determination of Cad vapor level with RGB color (red, green, blue) mode analysis. The test strips were employed to detect the freshness of real beef samples, and demonstrated a good capability of non-destructive, non-contact and visual screening meat freshness on site.

A chromo- and fluorogenic sensor for probing the cancer biomarker lysophosphatidic acid.

A novel chromo- and fluorogenic sensor, 4-(4-dimethylaminostyryl)-1-hexadecylpyridinium (DSHP) for lysophosphatidic acid (LPA) was successfully developed by incorporating a long alkyl chain into the cyanine molecule. DSHP shows excellent selectivity and high sensitivity towards LPA with a detection limit of about 7.09 × 10(-7) M based on electrostatic and hydrophobic interactions between the sensor and LPA. Upon addition of LPA ranging from 0 to 7.5 × 10(-4) M, DSHP displays an 'on-off-on' fluorescence response and obvious colour change. Good linear relationships between the fluorescence intensity and LPA concentrations were achieved in the fluorescence quenching ranges of 0-28 µM and 34-52 µM, which could be attributed to the combined effects of the photoinduced electron transfer and LPA-induced aggregation of the sensor molecules.

A smartphone-adaptable chromogenic and fluorogenic sensor for rapid visual detection of toxic hydrazine in the environment.

Hydrazine is an essential chemical in industries, but its high toxicity poses great threats to human health and environmental safety. Hence, it is of great significance to monitor the hydrazine in environment. In this work, we presented a chromogenic and fluorogenic dual-mode sensor RA for the detection of hydrazine based on nucleophilic substitution reaction. A linear relationship was obtained between the fluorescence intensity and the concentrations of N2H4 ranging from 0 to 35 µM (R2 = 0.9936). The sensor can determine hydrazine with fast response (within 12 min), low limit of detection (0.129 µM) and high selectivity. RA was successfully used to detect N2H4 in real water samples with good recoveries and the results corresponded to the standard method. Furthermore, the sensor-coated portable test papers were fabricated, which can visually quantify hydrazine solutions with obvious fluorescence transformation from colorless to red. Moreover, RA-loaded papers were used to create a smartphone-adaptable RGB values analytical method for quantitative N2H4 detection.

Fast visual monitoring of the freshness of beef using a smart fluorescent sensor.

Spoiled meat contains many pathogenic bacteria; hence, the intake of spoiled food can lead to various illnesses. To screen the freshness of food, in this study, we devised a ratiometric fluorescence sensor dicyanovinyl coumarin (CMDC) for the determination of cadaverine, an important biomarker for the spoilage of meat. CMDC underwent aza-Michael addition with cadaverine, exhibiting high sensitivity, fast response (50 s), and distinct fluorescence color transition. Test strips fabricated using CMDC showed a noticeable color change from red to green when exposed to cadaverine vapor. The test strips were successfully used to visually monitor the spoilage of beef based on the fluorescence color change. Furthermore, the as-developed test strip coupled with a smartphone provides a simple tool for consumers and suppliers to obtain information about meat quality.

Early diagnostic imaging of pneumonia with an ultra-sensitive two-photon near-infrared fluorescent probe.

The mortality rate of pneumonia increases significantly with the prolongation of illness. In the pursuit of a tool to accurately diagnose pneumonia in its early stages, we designed and synthesized a two-photon near-infrared fluorescent probe (DCQN) to identify increased concentrations of the inflammation marker SO2. The probe was found to specifically react with SO2 by undergoing a 1,4-addition reaction to generate near-infrared fluorescence with good sensitivity (6 s), a large Stokes shift (110 nm) and low detection limit (1.49 nM). DCQN has near-infrared emission as well as good two-photon performance, which can image exogenous and endogenous SO2 in cells and avoid interference from background fluorescence from cells. Furthermore, this study achieved accurate imaging of a pneumonia lesion site in deep tissues to provide a tool for the fluorescence diagnostic imaging of pneumonia in situ.