Construction of a dual-excitation ratiometric fluorescent probe for determining peroxynitrite levels in living cells and zebrafish.

Peroxynitrite (ONOO-) is a highly reactive oxygen species that plays a critical role in many physiological and pathological processes of cell function. This study aimed to propose a ratiometric fluorescent probe BDHCA derived from coumarin for determining the ONOO- level. ONOO- could specifically induce oxidative cleavage of the conjugated C = C double bond in probe BDHCA, providing a fluorescent ratiometric output. The response of probe BDHCA to ONOO- was selective, fast, and highly sensitive, with a detection limit of 50.3 nM. Biological imaging experiments suggested that probe BDHCA could be used to image ONOO- in living RAW264.7 cells and zebrafish.

A dihydro-benzo[4,5]imidazo[1,2-c]quinazoline-based probe with aggregation-induced ratiometric emission for the ratiometric fluorescent detection of peroxynitrite in living cells and zebrafish.

As a significant kind of reactive oxygen species (ROS), peroxynitrite (ONOO-) plays an indispensable role in many physiological and pathological processes. This study aimed to synthesize a novel dihydro-benzo[4,5]imidazo[1,2-c]quinazoline-based probe 1 for detecting ONOO-. In 99.5% H2O solution, probe 1 displayed a distinct aggregation-induced ratiometric emission (AIRE), and would selectively respond toward ONOO-via a ratiometric fluorescent signal, along with a short response time (<30 s) and ultra-sensitivity (LOD = 17.6 nM). Moreover, the probe was applied for monitoring the concentration fluctuations of ONOO- in HeLa cells and zebrafish.

A zeolitic imidazolate framework-90-based probe for fluorescent detection of mitochondrial hypochlorite in living cells and zebrafish.

An inorganic-organic hybrid probe MP-ZIF-90 was synthesized via a simple condensation reaction based on the free CHO groups of zeolitic imidazolate framework-90 (ZIF-90) and 4-methyl-1-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl)pyridinium bromide (MP). This probe exhibited intense green emission, which was selectively quenched by the addition of ClO- anions. The response of probe MP-ZIF-90 toward ClO- was rapid (within 20 s) and sensitive, with a limit of detection (LOD) of 0.612 µM. Importantly, the utilization of the probe in the fluorescence imaging of ClO- anions in the mitochondria of living cells and zebrafish was demonstrated.

A coumarin-pyrazole-based probe for the fluorescence detection of phosgene with high selectivity and sensitivity.

The high toxicity of phosgene poses potential threats to public health and safety. In this work, a novel fluorescent probe was designed to detect phosgene using hydroxyl and pyrazole moieties as the recognition sites. The response to phosgene with probe 1 was fast (less than 30 s), highly selective and sensitive with the limit of detection being 4.78 nM in solution. Furthermore, probe 1 was employed to conveniently fabricate paper test strips for efficiently detecting phosgene gas. The limit of detection was obtained as 0.014 ppm by using a smartphone RGB app, revealing that probe 1 has good prospects for sensitively detecting phosgene gas.

A p-toluenesulfonamide-modified benzo[h]chromene hydrazone: Fluorescent turn-on detection of hypochlorite and its application to imaging the endoplasmic reticula of living cells and zebrafishes.

Hypochlorite (ClO-) is a ROS that plays a crucial role in the immune system in the body. As the largest organelle in the cell, the endoplasmic reticulum (ER) manages various life activities. Thus, a simple hydrazone-based probe was designed, which provided a fast turn-on fluorescent response toward ClO-. With a terminal p-toluenesulfonamide group as the endoplasmic reticulum (ER)-specific site, probe 1 was mainly accumulated at ER of living cells, and could be used for imaging endogenous and exogenous HClO in cells and zebrafishes.

A dual-emission fluorescence probe for the detection of viscosity and hydrazine in environmental and biological samples.

The microenvironments of biological systems are associated with the pathology of organisms. This study, aimed to construct a hemicyanine-based probe (1), which can respond to mitochondrial viscosity and hydrazine (N2H4), for imaging application in living cells and zebrafish. The probe showed no fluorescence due to the intramolecular rotation in the solution; however, it exhibited a strong emission at 730 nm when the molecules were restricted to a high-viscosity environment. The addition of N2H4 caused an elimination reaction of the N-substituted group in the pyridinium part and further broke the CC bond to produce a highly fluorescent hydrazone. Also, the probe could selectively and quantitatively detect N2H4 via the fluorescence enhancement at 510 nm in a concentration range of 0 µM-140µM, with the limit of detection being 0.0485 µM. This probe may be used to study diseases related to N2H4 and viscosity changes in biological systems. Furthermore, the analysis methods based on probe 1 for N2H4 detection in soil, water, and air samples were successfully established.

A dual-ratiometric mitochondria-targeted fluorescent probe to detect hydrazine in soil samples and biological imaging.

Hydrazine (N2H4) is carcinogenic, extremely toxic, and induces serious environmental contamination and physiological dysfunction; however, it is widely used as an industrial material. Hence, the development of a simple and effective analytical method to detect N2H4 detection in both environmental and biological sectors is warranted. In this work, an intramolecular charge transfer (ICT)-based fluorescent probe 1, namely (Z)- 1-(4-acetoxybenzyl)- 4-(1-cyano-2-(7-(diethylamino)- 2-oxo-2 H-chromen-3-yl)vinyl)pyridin-1-ium, was designed for dual-excitation (420 and 600 nm, excitation separations >160 nm), near infrared (NIR)-emissive, and ratiometric fluorescent detection of N2H4. The sensing behavior of probe 1 for N2H4 detection was shown to be available over a wide pH range, and detection limits of 68 nM and 569 nM were achieved at excitation wavelengths of 420 and 600 nm, respectively. In addition, probe 1 was successfully used to image mitochondrial N2H4 in living cells and zebrafish. Furthermore, the probe was also capable of determining hydrazine signals in test strips and environmental soil.

A simple colorimetric and fluorometric probe for rapid detection of CN- with large emission shift.

In this work, a novel probe R was synthesized via Knoevenagel reaction between 3H-benzo[f]chromium-2-formaldehyde and ethyl cyanoacetate for selective detection of CN- in both colorimetric and fluorescent signal channels. The recognition of CN- was through the nucleophilic reaction of CN- to C = C of probe R, which destroys π-conjugation and blocks the ICT effect of the probe, resulting in colorimetric and fluorometric responses. Probe R showed great sensitivity toward CN-, with large fluorescent emission (595 nm) and low detection limit (0.70 µM). Moreover, probe R can detect exogenous CN- in living cells.

Europium (III) complex-based fluorescent probe for instantaneous, selective, and sensitive detection of phosgene.

Phosgene (carbonyl chloride, COCl2) is a widely used colorless gas in organic synthesis. However, its high toxicity sets a severe potential damage of public safety. As the fluorescence method has the advantages of simple operation and real-time detection of phosgene, it is extremely important to develop a fluorescent phosgene probe for public health and safety. This study aimed to present a simple Eu3+ complex (1) with 2-hydroxyl-1H-benzimidazole moiety as a novel phosgene probe. Probe 1 exhibited characteristic emission of Eu3+ in CH3CN solution, which was specifically quenched after encountering phosgene. The change in the solution color from light red to dark could be easily distinguished with the naked eye under a 365 nm ultraviolet lamp. Finally, the test paper with probe 1 was fabricated for effortless, selective, and visual detection of phosgene gas.

Simultaneous detection of lysosomal SO2 and viscosity using a hemicyanine-based fluorescent probe.

The changes in sulfur dioxide and viscosity of lysosomes are significant indicators in physiological processes and the cell microenvironment. This study aimed to synthesize a hemicyanine-based probe for simultaneous detection of SO2 and viscosity. The probe could not only rationally detect sulfur dioxide in a semi-aqueous solution with high sensitivity (limit of detection = 0.78 µM) and fast response (within 30 s) but also monitor viscosity via fluorescence emission enhancement at 580 nm. Further, the dual-response probe was successfully used to image SO2 and viscosity in the lysosomes of living cells.

A novel indene-chalcone-based fluorescence probe with lysosome-targeting for detection of endogenous carboxylesterases and bioimaging.

An indene-chalcone-based fluorescence probe 1 was synthesized and characterized. Under physiological conditions (containing 5% DMSO), probe 1 showed satisfactory stability with a low background signal and recognized carboxylesterases (CEs) based on the catalytic hydrolysis of ester groups, releasing a significant green fluorescence. Probe 1 presents several features including a short response time (within 20 min), low detection limit (1.3 × 10-4 U/mL) and large stokes shift (over 155 nm). Notably, commercial lysosomal dye co-staining experiments illustrated the lysosomal localization function of 1, with the probe also being used for cell and zebrafish imaging of endogenous CEs.

A quinoline-based probe for the ratiometric fluorescent detection of sulfite in lysosomes of living cells.

A lysosome-targeting ratiometric fluorescent probe was synthesized for detecting sulfite based on sulfite-triggered nucleophilic addition reaction. Due to the specific reaction, the fluorescence intensity ratio (I530/I390) of the probe in an almost aqueous solution (0.5% DMSO) changed significantly after the addition of HSO3-, corresponding to the change in the fluorescence color of the solution from green to blue. The recognition was conducted using high-resolution mass spectrometry, proton nuclear magnetic resonance, and density functional theory calculations. The fluorescent probe could be utilized to quantitatively monitor HSO3- in lysosomes of living C6 glioma cells and real-water samples.

A ratiometric fluorescent probe for hydrogen sulfide in neat aqueous solution and its application in lysosome-targetable cell imaging.

Hydrogen sulfide (H2S) has been recently regarded as one of the most important gasotransmitters in the metabolic system, while abnormal H2S concentration is associated with various diseases. Although numerous fluorescent probes have been developed for the detection of cellular H2S, only a few of them can monitor lysosomal H2S with ratiometric fluorescent output. Here, we developed a water-soluble probe 1 toward H2S by introducing 2,4-dinitrophenyl ether into a novel merocyanine-based dye. As expected, H2S induced an obvious red-shift of the probe from 520 nm to 580 nm in neat aqueous solution, and this fluorescent ratiometric response is highly selective and sensitive (with the detection limit of 0.81 nM), rapid (within 10 s), and effective in a wide pH range (2.0-10.0). In particular, the probe was successfully applied for tracing H2S in the lysosomes of living cells and in zebrafish.

A deep-red lysosome-targetable fluorescent probe for detection of hypochlorous acid in pure water and its imaging application in living cells and zebrafish.

Hypochlorite plays a significant role in physiological processes, particularly regulation of lysosomal functions, and is involved in various diseases. Thus, it is crucial to develop highly sensitive and selective molecule tools to detect HClO in lysosomes. Herein, a novel 2H-benzo[h]chromene-pyridine derivative (1) was synthesized through condensation reaction, which exhibited a notable deep-red emission at 640 nm in pure water. This deep-red emission was specifically quenched by adding ClO-. The response of probe 1 toward ClO- was rapid (within 10 s), sensitive (detection limit of 0.012 µM), and effective over a wide range of pH (1.0-12.0). Due to the existence of morpholine as the lysosome-targeting unit, the probe was successfully utilized to monitor lysosomal ClO-. Moreover, the probe 1 was also applied to detecting ClO- in zebrafish.

A simple hydrazone probe for recognition of Al3+ and PPi and its applicability in lysosomal imaging.

A simple hydrazone probe (1) was designed and synthesized for the successive detection of Al3+ and pyrophosphate (PPi) in almost 100% buffer environment. The probe provided O2N donor set for chelation with Al3+, leading to a distinct fluorescence boost at 510 nm. The in-situ formed 1-Al3+ complex detected PPi with an "on-off" behavior. The detection limits for Al3+ and PPi were 35.7 nM and 76 nM, respectively. Benefiting from the existence of morpholine as lysosome-targeting group, probe 1 was successfully applied to the detection of Al3+ and PPi in lysosomes.

An AIRE-active far-red ratiometric fluorescent chemosensor for specifically sensing Zn2+ and resultant Zn2+ complex for subsequent pyrophosphate detection in almost pure aqueous media.

A simple Schiff-base fluorescent chemosensor (1) was synthesized by the reaction of 3-amino-pyrazine-2-carbohydrazide and 7-diethylamino-3-formylcoumarin; the sensor 1 displayed a notable green emission at 524 nm in DMSO and an aggregation-induced ratiometric emission (AIRE) at 555 nm in an almost buffered aqueous media (0.5% DMSO content). The AIRE of 1 was quenched following binding to Zn2+ ions, while the fluorescence emission in the far-red region was evidently enhanced at 628 nm. Notably, the ratiometric signal output could be utilized to specifically distinguish Zn2+ among various metal ions. Moreover, the 1-Zn2+ complex was effectively employed as a fluorescent ratiometric chemosensor for pyrophosphate (PPi) detection. The detection limit was 3.52 µM and 2.45 µM for Zn2+ and PPi, respectively. The binding mechanism was evaluated by 1H NMR, ESI-MS, single-crystal X-ray diffraction, TEM, time-resolved fluorescence spectrophotometry, and density functional theory studies. Overall, owing to its sensitive fluorescence behavior, cell imaging studies demonstrated that this sensor is capable of sensing Zn2+ and PPi in living cells.

Hydrazone derivative bearing coumarin for the relay detection of Cu2+ and H2S in an almost neat aqueous solution and bioimaging in lysosomes.

A new morpholine functionalized coumarin-based fluorescent probe 1 was easily synthesized. The probe realized the sequentially detecting of Cu2+ and H2S in the HEPES buffer solution (20 mM, pH = 5.0). It made a turn-off fluorescence response to Cu2+ by using a complex formation with a 2:1 binding mode, and the resulting complex was able to detect H2S according to the displacement approach with a turn-on fluorescence response. The detecting limits of probe 1 for Cu2+ and 1-Cu2+ system for H2S were calculated to be 26 nM and 88.5 nM, respectively. This "on-off-on" recognition process was demonstrated by ultraviolet-visible spectroscopy, fluorescence spectroscopy, using proton nuclear magnetic resonance studies, electrospray ionization mass spectroscopy, single crystal X-ray diffraction, and using density functional theory calculations. In addition, both cell imaging and co-staining experiments showed that the probe could be utilized to visually detect Cu2+ and H2S in lysosomes.

Effect of graphene oxide with different morphological characteristics on properties of immobilized enzyme in the covalent method.

Although graphene oxide (GO) has great potential in the field of immobilized enzyme catalysts, the detailed effects of GO with different morphological structures on immobilized enzyme are not well understood. GOs were prepared from 8000 mesh and nanoscale graphite at different reaction temperatures, and used as carriers to immobilize alpha-amylase by cross-linking method. The properties of GOs were characterized through Atomic force microscope, Fourier-transformed infrared, X-ray photoelectron spectroscopy, Raman and UV-Vis. Furthermore, the dosage of cross-linking agent, cross-linking time, optimum temperature/pH, thermal/pH/storage stability, reusability and kinetic parameters of immobilized enzymes were investigated. The results showed that the loading of alpha-amylase on GOs was 162.3-274.2 mg g-1. The reusability experiments revealed high activity maintenance of immobilized alpha-amylase even after seven reaction cycles. Moreover, the storage stability of immobilized enzyme improved via immobilization in comparison with free one and it maintained over 70% of their initial activity after 20 days storage at 4 °C.

Sensitive and selective fluorescent probe for hypochlorite in 100% aqueous solution and its application for lysosome-targetable cell imaging.

A morpholine-functionalized pyrrole-cyanine probe was synthesized via a simple condensation reaction in high yield. This probe exhibits high selectivity toward ClO- on fluorescence and UV-vis spectra in neat aqueous solution. The strong green emission of the probe solution was quenched and the yellow color faded immediately upon the addition of ClO-. The detection limit of the probe for ClO- was 0.165 µM. The mechanism of hypochlorite-induced CC breakage was supposed on the basis of EIS-MS, NMR, and density functional theory (DFT) calculation. Finally, the probe was utilized to image ClO- in lysosomes of living cells.

Pyrrole-quinazoline derivative as an easily accessible turn-off optical chemosensor for Cu2+ and resultant Cu2+ complex as a turn-on sensor for pyrophosphate in almost neat aqueous solution.

A simple chemosensor, 6-(1H-pyrrol-2-yl)-5,6-dihydro-benzo[4,5]imidazo[1,2-c]quinazoline (1), was synthesized via simple nucleophilic addition reaction coupled with Schiff base condensation. The probe 1 is aggregation-induced emission-active and could be used as an on-off fluorescence sensor toward Cu2+ in H2O/CH3CN (99.5%, v/v) solution. Furthermore, the resultant Cu2+ complex selectively responded to pyrophosphate (PPi) among various anions based on fluorescent on-off signal. In addition, the probe could be used for detecting Cu2+ and PPi in HeLa cells.