A real-time ratiometric fluorescent probe for imaging of SO2 derivatives in mitochondria of living cells

A real-time ratiometric fluorescent probe (IN-CZ) for highly selective detection of sulfite was designed and synthesized, which is based on modulating the intramolecular charge transfer (ICT) of the hemicyanine dye platform. The mechanism of using the probe is mainly through the Michael addition that occurs between IN-CZ and sulfite with a detection limit of 2.99 × 10⁻⁵ M. IN-CZ displays a fast response (within 1 minute) and is highly selective for SO₃²⁻/HSO₃⁻ over ROS, biologically relevant ions, biological mercaptans and other reactive species. More importantly, IN-CZ was suitable for ratiometric fluorescence imaging in living cells, by real-time monitoring of SO₃²⁻/HSO₃⁻ changes in mitochondria targeted in living cells.

A real-time ratiometric fluorescent probe (IN-CZ) for highly selective detection of sulfite was designed and synthesized, which is based on modulating the intramolecular charge transfer of the hemicyanine dye platform.     

A ratiometric fluorescent probe for detection of exogenous mitochondrial SO2 based on a FRET mechanism

A novel imidazo[1,5-a]pyridine-hemicyanine based ratiometric fluorescent probe for detection of mitochondrial SO₂ was designed and synthesized. The probe is based on a fluorescence resonance energy transfer (FRET) mechanism. It exhibits high selectivity and sensitivity towards SO₃²⁻ with a fast response time (3 min) and detection limit of 0.13 μM. Further, it showed low cytotoxicity and was successfully applied to image exogenous mitochondrial SO₂ in cells.

A novel imidazo[1,5-a]pyridine-hemicyanine based ratiometric fluorescent probe for detection of mitochondrial SO₂ was designed and synthesized.     

A sensitive and selective BINOL based ratiometric fluorescence sensor for the detection of cyanide ions

A highly selective, novel BINOL based sensor BBCN has been developed for the fluorescent ratiometric detection of cyanide ions (CN⁻). The optical study revealed that BBCN exhibited unique spectral changes only with cyanide ions in the presence of other competing ions. Besides, an apparent fluorescent colour change from green to blue was observed. A clear linear relationship was observed between the fluorescence ratiometric ratio of BBCN and the concentration of CN⁻ with a reasonably low detection limit (LOD) of 189 nM (507 ppb). The optical response was due to the nucleophilic addition of CN⁻ to the dicyanovinyl group of the sensor, which compromises the probe''s intramolecular charge transfer. This mechanism was well confirmed by Job''s plot, ¹H-NMR and ESI-MS studies. BBCN showed immediate spectral response towards (1 second) CN⁻ and detection could be realized in a broad pH window. Furthermore, the practical utility of BBCN was studied by test paper-based analysis and the detection of CN⁻ in various water resources.

A highly selective, novel BINOL based sensor BBCN has been developed for the fluorescent ratiometric detection of cyanide ions (CN⁻).     

One-step synthesis of green emission carbon dots for selective and sensitive detection of nitrite ions and cellular imaging application

In recent years, carbon dot (CD)-based fluorescent sensors for selective ions or small biomolecules have drawn great attention. In this work, highly fluorescent CDs (QY = 21%) were prepared from 2,3-diamino pyridine as the precursor through a facile solvothermal process. The CDs showed high stability and a green emission in aqueous, and the optimal emission wavelength of CDs is 508 nm under the excitation wavelength of 438 nm. Interestingly, a CDs-based nanoprobe was developed for a selective and sensitive fluorescence quenching response to NO₂⁻ in water, and the quenching mechanism was investigated in the work. Besides, the recovery rates of NO₂⁻ in the range of 98–103.5% were found to be acceptable, indicating that the proposed CDs could be act as potential candidates for determination of nitrite ions in real samples. Meanwhile, the nanoprobe was also successfully employed in a visualization biosensing platform for determination of NO₂⁻ in living cells due to its eminent biocompatibility.

Schematic route of the carbon dots and their applications for the nitrite detection.     

Preparation of N/S doped carbon dots and their application in nitrite detection

Detection of carcinogens is generally recognized to be meaningful, especially for nitrites (NO₂⁻). Here blue-green fluorescent carbon dots (CDs) were successfully synthesized by using p-aminobenzenesulfonic acid, and their surfaces were identified to be abundant in the functional groups of amino, hydroxyl, and sulfuric acid. Importantly, the sulfuric acid group and aromatic primary ammonia groups on the surfaces of CDs showed the interactions with the nitrites to cause fluorescence quenching. The novel CDs showed high sensitivity and selectivity for NO₂⁻ detection with a low detection limit of 0.03 mM in water due to the fluorescence quenching effect of the CDs. Consequently, the proposed CDs here may provide a new way of monitoring NO₂⁻ in the target samples.

Detection of carcinogens is generally recognized to be meaningful, especially for nitrites (NO₂⁻).     

A dual emission metal–organic framework for rapid ratiometric fluorescence detection of CO32− in seawater

A dual emission metal–organic framework (IRMOF-10-Eu) was prepared and used as a ratiometric fluorescent sensor for CO₃²⁻ detection. IRMOF-10-Eu had good stability and excellent luminescence in aqueous solution. IRMOF-10-Eu showed dual fluorescence emission from the ligand and Eu³⁺ with single excitation. Upon treatment with CO₃²⁻, the fluorescence ratio (I₆₂₄/I₃₅₈) of the probe displayed significant change. The relative fluorescence intensity ratio (I₆₂₄/I₃₅₈) and CO₃²⁻ concentration had a linear relationship in 50–300 μM range with a low detection limit of 9.58 μM. And the luminescence probe of CO₃²⁻ showed a fast detection time. The possible mechanism was investigated. CO₃²⁻ changed the structure of IRMOF-10-Eu and interrupted the energy transfer process. Thus, the fluorescence emission intensity of the ligand was increased and Eu³⁺ was decreased with the addition of CO₃²⁻. IRMOF-10-Eu was used to detect CO₃²⁻ in seawater, which showed good prospect in practical application. Subsequently, a highly selective and sensitive probe, IRMOF-10-Eu, may pave an efficient way for CO₃²⁻ detection in seawater.

A dual emission metal–organic framework (IRMOF-10-Eu) was prepared and used as a ratiometric fluorescent sensor for CO₃²⁻ detection.     

A methylene blue-based near-infrared fluorescent probe for rapid detection of hypochlorite in tap water and living cells

A methylene blue-based near-infrared fluorescent probe was designed for the selective determination of hypochlorite (ClO⁻), over other reactive oxygen species or interfering agents. Acetylated methylene blue was synthesized by introducing the acetyl group into the methylene blue framework, which can specifically recognize exogenous and endogenous ClO⁻. The acetylated methylene blue fluorescent probe was characterized by ¹H NMR, ¹³C NMR and HRMS. The response process and possible mechanism were studied using products of the probe. The emission response of the probe to ClO⁻ presented good linear relationship in the 0–60 μM concentration range, with the detection limit of 0.1 μM (measured at 660 nm and 690 nm). The absorption and emission wavelengths of acetylated methylene blue are both in the near-infrared region; in addition, the probe itself and the degradation products were well-dissolved in water and have almost no toxicity. The probe was used for intracellular ClO⁻ imaging and showed a large fluorescence enhancement (about 200-fold increase).

We developed a MB-based probe to detect OCl⁻, whose product is almost non-toxic. The fluorescence enhancement times are large.     

A dual responsive fluorescent probe for selective detection of cysteine and bisulfite and its application in bioimaging

A coumarin-based dual responsive fluorescent probe with a simple structure was developed for the detection of Cys and HSO₃⁻. Under simulated physiological conditions, Cou-F displayed an on–off fluorescence response to Cys at 521 nm and an off–on fluorescence response to HSO₃⁻ at 500 nm. Furthermore, Cou-F had the advantages of high sensitivity, strong specificity and rapid response. The detection limits of Cou-F toward Cys and HSO₃⁻ were 0.54 μM and 0.65 μM, respectively. Cou-F enabled high selective responses to Cys and HSO₃⁻ over other biologically related species. The response times of Cou-F toward Cys and HSO₃⁻ were 80 s and 100 s. The fluorescence imaging of Cys and HSO₃⁻ was achieved in living RAW246.7 cells.

A coumarin-based dual responsive fluorescent probe with a simple structure was developed for the detection of Cys and HSO₃⁻.     

A gold nanoparticle based colorimetric and fluorescent dual-channel probe for acetylcholinesterase detection and inhibitor screening

Based on the competitive host–guest interaction between p-sulfonatocalix[6]arene (p-SC₆A) capped AuNPs and Rhodamine B (RhB)/acetylthiocholine, a fluorescent and colorimetric dual channel probe was developed for rapid detection of AChE with high sensitivity and selectivity. The detection limit was estimated to be 0.16 mU mL⁻¹. Crucially, due to the specific host–guest interaction, the high selectivity of the bioassay permitted the discrimination of AChE from other cations and proteins including biothiols and enzymes. Furthermore, the present method was also successfully applied to determinate AChE levels and screen AChE inhibitors in real cerebrospinal fluid (CSF) samples, which suggested that our proposed method has great potential to be applied in monitoring the disease progression and drug treatment effects of Alzheimer''s disease (AD).

A novel colorimetric and fluorescent dual-channel probe was developed for acetylcholinesterase detection and inhibitor screening with high sensitivity and selectivity.     

A coumarin Schiff's base two-photon fluorescent probe for hypochlorite in living cells and zebrafish

Selective and sensitive fluorescent probes for ClO⁻ are desirable due to the importance of ClO⁻ in biological processes. Here, a coumarin Schiff''s base, compound 1, has been developed and successfully used as a one- and two-photon fluorescent probe for ClO⁻ with high selectivity. This probe can recognize ClO⁻ with obvious color change from yellow-green to colorless and green to blue fluorescence emission, which can be observed by the naked eye. The properties of low cytotoxicity and good cell permeability allow it to be used for ClO⁻ detection in living cells and zebrafish by both one- and two-photon microscopy imaging. All these results indicate that the compound is a sensitive probe with potential for analysis of ClO⁻ in biological samples. The mechanism by which probe 1 recognizes ClO⁻ is possibly nucleophilic addition followed by hydrolysis.

A coumarin Schiff''s base compound can selectively recognize ClO⁻ and can be successfully applied to the detection of ClO⁻ in living cells and zebrafish by one- and two-photon fluorescence modes.     

Ratiometric fluorescence probe of Cu2+ and biothiols by using carbon dots and copper nanoclusters

A novel dual-emission ratiometric fluorescent probe based on N-doped yellow fluorescent carbon dots (y-CDs) and blue fluorescent copper nanoclusters (CuNCs) was established for quantitative determination of Cu²⁺ and biothiols. In this work, the Cu²⁺-(y-CDs) complexes formed by the chelation of y-CDs with Cu²⁺, showed an absorption peak at 430 nm that not only enhanced the fluorescence of y-CDs through inhibiting photoinduced electron transfer (PET) but also effectively quenched the fluorescence of CuNCs due to Förster resonance energy transfer (FRET). In addition, the chelation of y-CDs with Cu²⁺ could be inhibited by biothiols that prevented the fluorescence of y-CDs from being enhanced and the fluorescence of CuNCs from being quenched. On account of the changes of ratiometric signal, a dual-emission fluorescence probe for Cu²⁺ and biothiols determination was achieved. The proposed method exhibited high sensitivity for Cu²⁺ and biothiols in the ranges of 0.5–100 μM and 0.8–50 μM and the limits of detection (LODs) of Cu²⁺, glutathione (GSH), cysteine (Cys) and homocysteine (Hcy) were 0.21 μM, 0.33 μM, 0.39 μM and 0.46 μM, respectively. Subsequently, the established strategy presented an application prospect for the detection of Cu²⁺ and biothiols in real samples.

A dual-mechanism ratiometric fluorescent probe based on N-doped yellow fluorescent carbon dots (y-CDs) and blue fluorescent copper nanoclusters (CuNCs) was established for the first simultaneous determination of Cu²⁺ and biothiols.     

A new ratiometric fluorescence assay based on resonance energy transfer between biomass quantum dots and organic dye for the detection of sulfur dioxide derivatives

Sulfur dioxide (SO₂) is considered as the fourth gas signal molecule after nitric oxide (NO), carbon monoxide (CO) and hydrogen sulfide (H₂S). It plays important roles in several physiological processes. Therefore, the design and synthesis of nanoprobes for the detection of SO₂ derivatives in cells is of great significance. Herein, we report a new ratiometric fluorescence nanoprobe based on resonance energy transfer (RET) between biomass quantum dots (BQDs) and organic dye (DMI) for the detection of SO₂ derivatives. The proposed ratiometric fluorescence assay allows the determination of HSO₃⁻ in the range of 1.0 to 225 μM with a detection limit of 0.5 μM. Importantly, the proposed ratiometric fluorescence nanoprobe exhibits a high photostability and good selectivity for HSO₃⁻ over other chemical species including H₂S and biological mercaptans. Quantitation of HSO₃⁻ in cell lysates by using the nanoprobe is demonstrated.

A new ratiometric fluorescence assay has been developed for the detection of sulfur dioxide derivatives with repeatability and selectivity. The assay was applied to quantitate HSO₃⁻ in cell lysates with accurate results.     

A naphthalimide-based turn-on fluorescence probe for peroxynitrite detection and imaging in living cells

Peroxynitrite (ONOO⁻) is a potent biological oxidant that plays a significant role in diverse physiological and pathological processes. A novel fluorescent probe HCA-OH was developed for specific and sensitive detection of peroxynitrite, and displayed a significant fluorescence turn-on signal. With low cytotoxicity and good photostability, the probe HCA-OH could be applied in imaging ONOO⁻ distribution in HepG2 cells and live C. elegans in real time. Therefore, the probe can be a promising tool for imaging in vivo.

A novel fluorescent probe HCA-OH was designed for selective detection of peroxynitrite and imaging in HepG2 cells and C. elegans.     

Orange emissive carbon dots for colorimetric and fluorescent sensing of 2,4,6-trinitrophenol by fluorescence conversion

In this study, infrequent orange carbon nanodots (CNDs) were applied as a dual-readout probe for the effective colorimetric and fluorescent detection of 2,4,6-trinitrophenol (TNP). The orange fluorescence could be rapidly and selectively quenched by TNP, and the colorimetric response from the original pink color to blue could also be captured immediately by the naked eye. A limit of detection of 0.127 μM for TNP was estimated by the fluorescent method and 5 × 10⁻⁵ M by visualized detection. Interestingly, the fluorescence of the CNDs with TNP gradually transitioned from orange to green upon irradiation by a UV lamp, and the colorimetric response transitioned from pink to blue to colorless, which ensured effective multi-response detection of TNP. In addition, the CNDs exhibited bright fluorescence, excellent biocompatibility and low toxicity, making them high-quality fluorescent probes for cellular imaging.

We have described a colorimetric and fluorescent dual-readout probe with a strong and sensitive response towards TNP.     

A highly specific and sensitive turn-on fluorescence probe for hypochlorite detection and its bioimaging applications

Development of high performance fluorescent chemosensors for the detection of ClO⁻in vitro and in vivo is very desirable, because many human diseases are caused by ClO⁻. In this paper, a highly selectivity and sensitive fluorescent probe, EDPC, based on 3-acetylcoumarin, was synthesized, which could respond to ClO⁻ and exhibit an “off–on” mode in Tris–HCl buffer (pH = 7.2, 10 mM, 50% C₂H₅OH) solutions. The detection limit of the EDPC probe for ClO⁻ was as low as 1.2 × 10⁻⁸ M. Moreover, the high selectivity and high sensitivity of EDPC towards ClO⁻ are attributed to the oxidation reaction between the C–O of the coumarin lactone and the C C formed by aldol condensation and the mechanism was further verified using ESI-MS and DFT. Additionally, the concentrations of ClO⁻ in real water were also calculated using the EDPC probe and showed good recovery. Finally, the distribution of intracellular endogenous ClO⁻ was gained by confocal fluorescence microscopy in living HEK293T cells.

A new probe, EDPC, was designed and synthesized for highly specific and sensitive detection of ClO⁻ with a fast response time in living cells.     

Semi-quantitative and visual assay of copper ions by fluorescent test paper constructed with dual-emission carbon dots

A novel, simple and effective dual-emissive fluorescent probe for the sensitive and selective detection of Cu(ii) has been developed by mixing blue carbon dots and orange carbon dots, with a sensitive detection limit of 7.31 nM. The blue fluorescence can be selectively quenched by Cu(ii), while the orange fluorescence is a internal reference, resulting in a distinguishable fluorescence color change from blue to orange under a UV lamp. Meanwhile, its as-prepared text paper provides a convenient and simple approach for the visual detection of Cu(ii) and successfully applied in real water samples, with a dose-discerning ability as low as 50 nM. The methodology reported here opens a novel pathway toward the real applications of fluorescent test papers.

A ratiometric fluorescence nanosensor has been developed by mixing blue fluorescent carbon dots and orange fluorescent carbon dots for the detection of copper ions.     

Cobalt metal-mixed organic complex-based hybrid micromaterials: ratiometric detection of cyanide

A micrometer-sized hybrid crystalline material was facilely fabricated from the synergistic coordination chemistry of two types of 2-hydroxyazobenzene building blocks and cobalt ions. This hybrid crystalline material functioned as both receptor and signal reporter in cyanide (CN⁻) sensing with ratiometric absorption responses at 456 nm and 537 nm. The mechanism of CN⁻ sensing involves partial ligand dissociation accompanied by the formation of a new organic metal–CN⁻ adduct. We believe that this protocol would be valuable in achieving the expected selectivities and sensitivities for a wide variety of analytes in many chemical and biological systems in the future.

A cobalt metal-mixed organic complex-based hybrid micromaterial has been presented to detect CN⁻ with ratiometric absorption responses (A_{456 nm}/A_{537 nm}).     

An aggregation-induced emission-based fluorescence turn-on probe for Hg2+ and its application to detect Hg2+ in food samples

In this work, we presented a new tetraphenylethene-derived fluorescent probe TPE-M for Hg²⁺ detection in an aqueous solution. Probe TPE-M is molecularly dissolved in CH₃OH/PBS (20 mM, pH = 7.4) (3 : 7, v/v) mixed solution and is almost non-emissive. Reaction of TPE-M with Hg²⁺ leads to release of an AIE-active precursor 4, and results in a significant fluorescence enhancement. The Hg²⁺ recognition process has some distinct advantages including rapid response, high selectivity and sensitivity, strong anti-interference ability, and a low detection limit (4.16 × 10⁻⁶ M). Moreover, the probe is applicable to detect Hg²⁺ in real food samples such as shrimp, crab and teas, suggesting the practical applicability of TPE-M.

A new AIE-based fluorescent probe TPE-M for Hg²⁺ detection in an aqueous solution has been developed.     

Immunochromatographic assay for T-2 toxin based on luminescent quantum dot beads

A quantum dot bead based immunochromatographic assay (QB-ICA) system was established for T-2 toxin (T-2), which widely occurs in agriculture and could be used as a potential biological warfare agent. After optimization, the dynamic linear detection range of T-2 calculated from a calibration curve was from 0.12 to 0.67 ng mL⁻¹ and the limit of detection (LOD) was 0.08 ng mL⁻¹, which is lower than those of the ICA based on colloidal gold nanoparticles or a fluorescent material or an antibody-based biochip in other reports. The performance and practicability of the established ICA system were validated with a commercial ELISA kit and the two methods were comparable. The proposed QB-ICA for T-2 could be an alternative for rapid, sensitive, and quantitative on-site detection of this toxin in biosafety monitoring in agriculture and for susceptibility testing of the potential release of this biological warfare agent.

A quantum dot bead based immunochromatographic assay was established for T-2 toxin with a limit of detection of 0.08 ng mL⁻¹.     

A “Turn-On” fluorescent probe for sensitive and selective detection of fluoride ions based on aggregation-induced emission

Based on the fluorophore of 2-(2′-hydroxyphenyl)benzothiazole (HBT) with aggregation-induced emission (AIE) properties, a highly selective and sensitive fluorescent probe PBT towards F⁻ was investigated. “Turn-On” fluorescence type signaling was realized by employing fluoride-selective cleavage of the latent thiophosphinated probe in mixed aqueous media. The probe is designed in such a way that the excited state intramolecular proton transfer (ESIPT) of the HBT moiety becomes blocked. The chemodosimetric approach of F⁻ to the probe results in the recovery of the ESIPT by removal of a free AIE-active HBT moiety through a subsequent hydrolysis process. The F⁻ detection limit of the probe was 3.8 nM in the dynamic range of 0.5 μM to 10 μM. In addition, the proposed probe has been used to detect F⁻ in water samples and toothpaste samples with satisfying results.

A “Turn-On” fluorescent probe PBT for sensitive and selective detection of fluoride ions based on aggregation-induced emission.