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The eukaryotic cell is highly compartmentalized with organelles. Owing to their function in transporting metabolites, metabolic intermediates and byproducts of metabolic activity, organelles are important players in the orchestration of cellular function. Recent advances in optical methods for interrogating the different aspects of organellar activity promise to revolutionize our ability to dissect cellular processes with unprecedented detail. The transport activity of organelles is usually coupled to the transport of charged species; therefore, it is not only associated with the metabolic landscape but also entangled with membrane potentials. In this context, the targeted expression of fluorescent probes for interrogating organellar membrane potential (Ψ org ) emerges as a powerful approach, offering less-invasive conditions and technical simplicity to interrogate cellular signalling and metabolism. Different research groups have made remarkable progress in adapting a variety of optical methods for measuring and monitoring Ψ org . These approaches include using potentiometric dyes, genetically encoded voltage indicators, hybrid fluorescence resonance energy transfer sensors and photoinduced electron transfer systems. These studies have provided consistent values for the resting potential of single-membrane organelles, such as lysosomes, the Golgi and the endoplasmic reticulum. We can foresee the use of dynamic measurements of Ψ org to study fundamental problems in organellar physiology that are linked to serious cellular disorders. Here, we present an overview of the available techniques, a survey of the resting membrane potential of internal membranes and, finally, an open-source mathematical model useful to interpret and interrogate membrane-bound structures of small volume by using the lysosome as an example., (© 2024 The Authors. The Journal of Physiology © 2024 The Physiological Society.)

Background: Heavy metal pollution has become one of the world's most important environmental pollution, especially Hg 2+ is enriched, it is easy to enter the human body through the food chain, bind to the sulfhydryl group in the protein, cause mercury poisoning. Traditional methods for detecting Hg 2+ have obvious drawbacks, such as poor selectivity and long detection time. Fluorescence detection has attracted attention because of its good sensitivity and specificity detection ability. In previously reported probes for detecting Hg 2+ , Cu 2+ often interferes. Therefore, it is of great practical significance to synthesize a fluorescent probe that can distinguish between Hg 2+ and Cu 2+ ., Results: We have successfully synthesized the probe DFS, a fluorescent probe that can differentially detect Hg 2+ and Cu 2+ , and the probe DFS has good selectivity and anti-interference ability for Hg 2+ and Cu 2+ . The fluorescence intensity at 530 nm increased rapidly when Hg 2+ was detected; during the Cu 2+ detection, the fluorescence intensity at 636 nm gradually decreased, fluorescence quenching occurred, and the detection limits of Hg 2+ and Cu 2+ were 7.29 × 10 -9  M and 2.13 × 10 -9  M, respectively. Through biological experiments, it was found that probe DFS can complete the fluorescence imaging of Hg 2+ and Cu 2+ in Staphylococcus aureus and HUVEC cells, which has certain research value in the field of environmental monitoring and microbiology, and the probe DFS has low cytotoxicity, so it also has broad application prospects in the field of biological imaging. In addition, the probe DFS also has good applicability for Hg 2+ and Cu 2+ detection in actual samples., Significance and Novelty: This is a fluorescent probe that can distinguish between Hg 2+ and Cu 2+ , the fluorescence emission peak appears at 530 nm when Hg 2+ is detected; when detecting Cu 2+ , fluorescence quenching occurs at 636 nm, the fluorescence emission peak distance between Hg 2+ and Cu 2+ differs by 106 nm. This reduces mutual interference between Hg 2+ and Cu 2+ during detection, it provides a new idea for the detection of Hg 2+ and Cu 2+ ., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

In eutrophic water bodies, sulfides are closely related to the growth of cyanobacteria and the production of microcystin-LR (MC-LR). To date, the underlying interaction mechanism between a sulfides and MC-LR remains controversial. Thus, visually presenting the distribution characteristics of sulfides and MC-LR in contaminated water is crucial. Here, we propose a novel and expeditious practical approach, utilizing fluorescence probe technology, to assess the distribution characteristics of MC-LR and sulfur in natural lakes. We have developed novel probes, pib2, to detect HSO 3 - and HS - , and pib18, to simultaneously identify MC-LR and sulfides. Through correlation analysis of fluorescence data and physicochemical indicators at sampling points, it is found that fluorescence data has good correlation with sulfides and MC-LR, and speculated that pib2 and pib18 may be able to detect sulfides and MC-LR in lakes. Using this method, we rapidly obtained the distribution of MC-LR and sulfur in Qilu and Erhai Lakes. Notably, for the first time, we rapidly displayed the distributions of sulfides and MC-LR across lakes by the fluorescent probe technology., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

As cyanide is a huge hazard to the environment and human health, the study of the method of detecting low concentrations of cyanide is of great significance. In general, materials with strong positive electrostatic properties can use electrostatic attraction to enrich anions in the water near the materials, then realize rapid detection of low concentration anions by fluorescent probes. In this paper, fluorescent probes PI-S, PI-I and PI-N with cyanide-specific recognition and different charges were synthesized to study the relationship between the charge effect of probes and the sensing sensitivity. Through the zeta potential test and the calculation of the surface electrostatic potential, the positive electricity of PI-S, PI-I and PI-N gradually increased, the ΔG < 0 of the adsorption process gradually decreased, CN - could be aggregated to the vicinity of probes. As a result, the detection limit of the probe was gradually reduced from 1.07 × 10 -6 to 5.03 × 10 -8 M, the sensitivity was significantly enhanced. Therefore, this is expected to be a new strategy to improve the sensitivity of anion probes by increasing the positive electricity of molecules. In addition, PI-N has good anti-interference ability, short response time and certain application value in cell imaging and identification of endogenous cyanide in food., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Hydrazine (N 2 H 4 ) plays an important role in industrial production, but it is highly toxic, leaking or exposing it will pollute the environment and cause serious harm to human beings. Therefore, it is necessary to use a simple and effective method to detect N 2 H 4 in environmental systems and organisms. Herein, a novel water-soluble fluorescent probe based on coumarin fluorophore, 2-(7-(diethylamino)-2-oxo-2H-chromen-3-yl)isoindoline-1,3-dione (C-Z1), is reported. The fluorescence intensity of the probe at 530 nm was enhanced gradually with the addition of N 2 H 4 , and the maximum enhancement was about 28 times. The probe has good selectivity and sensitivity, the detection limit of hydrazine hydrate is 1.48 × 10 -7  M, and the response mechanism of the probe is proved by theoretical calculation and experiment. C-Z1 has been shown to detect N 2 H 4 in a variety of environmental samples, including water, soil, air, cells, zebrafish and plants. In addition, C-Z1 can be made into test strips for easy portability and used for rapid quantitative detection of N 2 H 4 in the field by its distinct change in fluorescence color. Thus, C-Z1 has great potential for the analysis and detection of environmental contaminants., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Atmospheric brown carbon (BrC) contain amounts of organic species, but their molecular weight (MW) distributions is still poorly understood. This study applied high-performance size exclusion chromatography (HPSEC) coupled with a diode array detector (DAD) and fluorescence detector (FLD) to characterize the MW distributions of typical chromophores and fluorophores within water-soluble BrC. The investigation focused on the spring season, encompassing both typical urban and rural aerosols. Our results showed that chromophores (at 254 and 365 nm), and humic-like and protein-like fluorophores identified by excitation-emission matrix parallel factor analysis (EEM-PARAFAC) within BrC were broadly distributed along the MW continuum (∼50-20,000 Da). This suggests that BrC mainly comprises complex chromophores and fluorophores with heterogeneous molecular sizes. High-MW (HMW, >1 kDa) species (66%-74%) dominated the chromophores at 254 and 365 nm. However, the latter chromophores were enriched with more HMW species. This result suggested that the HMW chromophores might contribute more to BrC absorption at longer wavelengths. The PARAFAC-derived fluorescent components also exhibited different MW distributions. Three humic-like substances (HULIS) were all dominated by HMW fractions (51%-74%), but protein-like fluorescent component (PLOM) enriched low-MW (LMW, <1 kDa) species (60%-66%). Furthermore, the molecular size (i.e., weight-averaged and number-averaged MW) and the ratios between HMW and LMW species decreased in the order highly-oxygenated HULIS > less-oxygenated HULIS > PLOM, indicating that the fluorophores with longer Em were generally related to larger MW. To our knowledge, this is the first report on the molecular size of individual fluorescent components within aerosol BrC. The results obtained here enhanced our knowledge of heterogeneous composition, complex physicochemical properties, and potential atmospheric fates of aerosol BrC., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

The widespread use of cyanide (CN - ) in industry results in contamination of various effluents such as drain, lake, and tap water, an imminent danger to the environment and human health. We prepared naphthoquinolinedione (cyclized; 1-5) and anthracenedione (un-cyclized) probes (6-7) for selective detection of CN - . The addition of CN - to the probe solutions (1-5) resulted in a color change from pale green to orange under 365 nm illumination. The nucleophilic addition of CN - to C2 of the imidazolium ring of the probes is responsible for selective CN - detection. Among all probes, 1 gave the lowest fluorescence-based LOD of 0.13 pM. In contrast, the un-cyclized probes (6 and 7) were substantially inferior to the cyclized counterparts (1 and 2, respectively) for detecting a trace amount of CN - . The notably low LOD displayed by probe 1 was maintained in the detection of CN - in real food samples, human fluids, and human brain cells. This is the first report studying imidazolium-bearing naphthoquinolinedione-based probes for CN - sensing in 100% water., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Although lipophilic membrane dyes (LMDs) or probes (LMPs) are widely used to label extracellular vesicles (EVs) for detection and purification, their labelling performance has not been systematically characterized. Through concurrent side scattering and fluorescence detection of single EVs as small as 40 nm in diameter by a laboratory-built nano-flow cytometer (nFCM), present study identified that (1) PKH67 and PKH26 could maximally label ∼60%-80% of EVs isolated from the conditioned cell culture medium (purity of ∼88%) and ∼40%-70% of PFP-EVs (purity of ∼73%); (2) excessive PKH26 could cause damage to the EV structure; (3) di-8-ANEPPS and high concentration of DiI could achieve efficient and uniform labelling of EVs with nearly 100% labelling efficiency for di-8-ANEPPS and 70%-100% for DiI; (4) all the four tested LMDs can aggregate and form micelles that exhibit comparable side scatter and fluorescence intensity with those of labelled EVs and thus hardly be differentiate from each other; (5) as the LMD concentration went up, the particle number of self-aggregates increased while the fluorescence intensity of aggregates remained constant; (6) PKH67 and PKH26 tend to form more aggregated micelles than di-8-ANEPPS and DiI, and the effect of LMD self-aggregation can be negligible at optimal staining conditions. (7) All the four tested LMDs can label almost all the very-low-density lipoprotein (VLDL) particles, indicating potential confounding factor in plasma-EV labelling. Besides, it was discovered that DSPE-PEG 2000 -biotin can only label ∼50% of plasma-EVs. The number of LMP inserted into the membrane of single EVs was measured for the first time and it was confirmed that membrane labelling by lipophilic dyes did not interfere with the immunophenotyping of EVs. nFCM provides a unique perspective for a better understanding of EV labelling by LMD/LMP., (© 2023 The Authors. Journal of Extracellular Vesicles published by Wiley Periodicals, LLC on behalf of the International Society for Extracellular Vesicles.)

Mercury ions are notoriously difficult to biodegradable, and its abnormal bioaccumulation in the human body through the food chain can cause various diseases. Therefore, the quantitative and real-time detection of Hg 2+ is very extremely important. Herein, we have brilliant designed and synthesized (E)-O-(4-(2-(3-(dicyanomethylene)-5,5-dimethylcyclohex-1-en-1-yl)vinyl)phenyl) O-phenyl carbonothioate (ICM-Hg) as a selective fluorescent probe for Hg 2+ detection in real samples and intracellular staining. ICM-Hg displayed high specificity toward Hg 2+ by activating the intramolecular charge transfer (ICT) process, resulting in distinguished color change from colorless to bright yellow along with noticeable switch on yellow fluorescence emission. The fluorescent intensity of ICM-Hg at 585 nm shows a well linear relationship in the range of Hg 2+ concentration (0-45 μM), and the detection of limit for Hg 2+ is calculated to be 231 nM. Promisingly, ICM-Hg can efficiently detect Hg 2+ in real samples including tap water, tea, shrimp, and crab with quantitative recovery as well as the intracellular fluorescence imaging., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Tannic acid (TA)-a natural product-is a polyphenol derivative that occurs in certain kinds of beverages. A large amount of TA could give rise to an unpleasant flavour and could negatively affect the human body by causing stomach irritation, abdominal pain, nausea, vomiting, and even death. Thus, the need exists for a simple TA detection procedure that meets specific criteria such as on-site analysis, portability, and affordability. Herein, we present a new TA assay, which is based on the fluorescent quenching effect of an efficient fluorophore, and which comprises a smartphone-integrated homemade reader system. The fluorescent polyethyleneimine-derivatised polymer (FP), a strong emitter at 510 nm, was synthesised with the aid of a facile sonication method. In the presence of Eu 3+ ions, TA quenches the fluorescence of the FP via electrostatic interaction. A smartphone was used to capture an image of the FP undergoing fluorescence for conversion to RGB values. The blue channel was chosen for further analysis because it offered the highest R 2 -value compared to the red and green channels. We verified these results using a commercial spectrofluorometer and calculated the limit of detection of this assay as 87 nM and 20 nM for the homemade reader and spectrofluorometer, respectively. The detection range for TA with the proposed assay is 0.16-66.66 μM. The application of the proposed method to real beverage samples for TA detection demonstrates its analytical applicability., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

A rapid, facile and ultrasensitive fluorescence sensing system based on beer-derived nitrogen, phosphorus co-doped carbon quantum dots (N, P-CQDs) for the detection of ascorbic acid (AA) in fruits was proposed. N, P-CQDs were successfully synthesized by one-step hydrothermal method, which afforded a high quantum yield (21.7 %), and showed the fluorescence with a maximum emission wavelength of 450 nm at an excitation wavelength of 370 nm. Further, N, P-CQDs were employed as an efficient sensor for ultrasensitive Fe 3+ -detection at concentrations ranging from 1-20 µM and 100-300 µM, respectively. N, P-CQDs@Fe 3+ showed a high sensitivity and selectivity for AA detection. A linear response range for AA was obtained from 1 to 200 µM with limit of detection of 0.84 µM was obtained for AA. The result of MTT test showed that N, P-CQDs exhibit low toxicity, providing fast, accurate and less toxic route for testing AA in the food analysis fields., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Hepatic lipid droplets (LDs) and peroxynitrite (ONOO - ) levels are closely related to nonalcoholic fatty liver disease (NAFLD). Additionally, some drug-induced liver injury (DILI) is often associated with ONOO - . Here, we constructed and screened the quasi-LDs-targeted and ONOO - -responsive fluorescent probe MBDP-Py + to investigate the interaction of NAFLD with DILI. By monitoring the upregulation of the ONOO - levels and the accumulation of LDs, MBDP-Py + was more sensitive and efficient than tissue staining and serum markers detection in evaluating the early toxicity of NAFLD and diagnosing the anticancer-DILI. More importantly, the sensitive enhancement of fluorescence signals demonstrated that in different stages of NAFLD, the dominant element of liver injury was different in the NAFLD combined with DILI mice models. As the degree of NAFLD deepens, the synergistic effect of the two will lead to more serious liver damage.

Detection of highly toxic thiophenols in biological or environmental systems is of great importance. Therefore, fast, reliable, and sensitive probes are needed to detect thiophenols. Herein, a novel triphenylamine conjugated dicyanoisophorone-based near infrared fluorescence probe is reported to determine trace thiophenol (PhSH) levels. The probe demonstrates a distinct "turn-on" fluorescence response to thiophenol among the tested analytes and its quantum yield (Φ) increases from 0.011 to 0.142. It has low cytotoxicity with cell viability of 90-100% up to 10.0 μM of the probe, a strong anti-interference capability, a large Stokes shift (150 nm), and a fast response time (<1 min). In addition, the probe exhibits a good linear response to PhSH over the range from 0 to 15.0 μM with a detection limit of 32.3 nM (R 2  = 0.9978). The detection process is also confirmed through HPLC. The practical applicability of the probe is proved by a smartphone platform, TLC kit, plant tissue imaging, soil assay, tap, and lake water analysis with good recovery values (92.3-117%), and concentration-dependent live cell bioimaging PhSH from 5.0 to 15.0 μM. Therefore, the present probe is a robust candidate for monitoring PhSH levels in biological and environmental systems., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Dynamic measurements of molecular machines can provide invaluable insights into their mechanism, but these measurements have been challenging in living cells. Here, we developed live-cell tracking of single fluorophores with nanometer spatial and millisecond temporal resolution in two and three dimensions using the recently introduced super-resolution technique MINFLUX. Using this approach, we resolved the precise stepping motion of the motor protein kinesin-1 as it walked on microtubules in living cells. Nanoscopic tracking of motors walking on the microtubules of fixed cells also enabled us to resolve the architecture of the microtubule cytoskeleton with protofilament resolution.

Qualitative and quantitative analysis of mercury at concentration levels as low as parts per billion (ppb) is a basic and practical concern. The vast majority of research in this field has centered on the development of potent chemosensor to monitor mercuric (Hg 2+ ) ions. Mercury exists in three oxidation states, + 2, + 1 and 0, all of which are highly poisonous. In this study, (N 1 E,N 2 E)-N 1 ,N 2 -bis(pyrene-1-ylmethylene)benzene-1,2-diamine (PAPM), a novel photoluminescent sensor based on pyrene platform was synthesized. Over the tested metal ions (Cd 2+ , Co 2+ , Cu 2+ , Mg 2+ , Mn 2+ , Ni 2+ , K + , Na + , Zn 2+ , Sr 2+ , Pb 2+ , Al 3+ , Cr 3+ and Fe 3+ ) the sensor responds only to Hg 2+ by showing high selectivity and sensitivity. After treatment with mercuric ions at room temperature, the luminescence intensity of probe was quenched at 456 nm. The quenching of fluorescence intensity of probe upon addition of mercury is due to the effect of "turn-off" chelation enhanced quenching (CHEQ) by the formation of 1:1 complex. The ESI-MS spectrum and the Job's experimental results confirm the formation of 1:1 complex between PAPM and Hg 2+ . The detection limit and association constant of sensor for mercury is computed using fluorescence titration data and were found to be 9.0 × 10 -8  M and 1.29 × 10 5  M -1 respectively. The practical application of sensor towards recognition of mercury(II) ions was explored through economically viable test strips and also using cell imaging studies., (© 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Sensitive high-throughput analytic methodologies are needed to quantify microplastic particles (MPs) and thereby enable routine monitoring of MPs to ultimately secure animal, human, and environmental health. Here we report a multiplexed analytical and flow cytometry-based high-throughput methodology to quantify MPs in aqueous suspensions. The developed analytic MPs-quantification platform provides a sensitive as well as high-throughput detection of MPs that relies on the material binding peptide Liquid Chromatography Peak I (LCI) conjugated to Alexa-fluorophores (LCI F16C -AF488, LCI F16C -AF594, and LCI F16C -AF647). These fluorescent material-binding peptides (also termed plastibodies) were used to fluorescently label polystyrene MPs, whereas Alexa-fluorophores alone exhibited a negligible background fluorescence. Mixtures of polystyrene MPs that varied in size (500 nm to 5 μm) and varied in labeled populations were analyzed and sorted into distinct populations reaching sorting efficiencies >90 % for 1 × 10 6 sorted events. Finally, a multiplexed quantification and sorting with up to three plastibodies was successfully achieved to validate that the combination of plastibodies and flow cytometry is a powerful and generally applicable methodology for multiplexed analysis, quantification, and sorting of microplastic particles., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

This paper reported an improved molecular beacon method for the rapid detection of aflatoxin B1 (AFB1), a natural mycotoxin with severe carcinogenicity. With the assistance of a complementary DNA (cDNA) chain, the molecular beacon which consists of a DNA aptamer flanked by FAM and BHQ1 displayed a larger fluorescent response to AFB1, contributing to the sensitive detection of AFB1. Upon optimization of some key experimental factors, rapid detection of AFB1 ranging from 1 nM to 3 μM, within 20 min, was realized by using this method. A limit of detection (LoD) of 1 nM was obtained, which was lower than the LoD (8 nM) obtained without cDNA assistance. This aptamer-based molecular beacon detection method showed advantages in easy operation, rapid analysis and larger signal response. Good specificity and anti-interference ability were demonstrated. This method showed potential in real-sample analysis.

Extensive use of palladium in many catalysts and catalytic converters causes a high degree of pollution of water and soil resources. Therefore, there is an urgent need to develop rapid and sensitive palladium probes. Herein, a novel "turn-on" near-infrared (NIR) fluorescence and colorimetric probe for Pd has been designed on the basis of the deallylation of the probe, followed by the release of NIR emissive fluorophore through the Tsuji-Trost reaction. The probe can selectively discriminate between the oxidation states of Pd 0 and Pd 2+ . Sensing results demonstrates that the probe has excellent selectivity, sensitivity, fast response time, NIR fluorescence, high biocompatibility, and low detection limit for the Pd detection over a series metal ion. The probe has been successfully applied in visualization of residual Pd content from water, soil, drug and living cell samples by fluorescence observation with the naked eye., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Herein, a novel relay peptide-based fluorescent probe DGRK was synthesized via solid phase peptide synthesis (SPPS) technology. DGRK exhibited excellent water-solubility, good stability, remarkably large Stokes shift (230 nm) and high selectivity response to Hg 2+ with a non-fluorescence complex DGRK-Hg 2+ formation via a 1:1 binding mode. Further studies indicated that the DGRK-Hg 2+ complex could act as a secondary probe for rapidly and sequentially detecting S 2- based on fluorescent "off-on" response, and without interference from a range of anions. The limit of detection (LOD) for Hg 2+ and S 2- were calculated to be 33.6 nM and 60.9 nM, respectively. In addition, The reversibility of interaction of confirmed that the continuous and reversible recognition behavior of Hg 2+ and S 2- by the probe DGRK, and could be cycled more than 5 times. In addition, DGRK could be successfully applied to the fluorescence imaging of Hg 2+ and S 2- in two living cells based on excellent cells permeability and low cytotoxicity. Meanwhile, DGRK was successfully used to create the low-cost and portable test strips for visual detection and rapid analysis under 365 nm UV lamp, and the test strips combined with a smartphone (RGB color) was successfully applied to the semi-quantitative analysis and monitoring of dynamic changes of Hg 2+ levels., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

The threat of tetracycline antibiotics to the environment and human health is attracting widespread attention. The development of morphological analysis and quantitative techniques of multiple tetracyclines is of great significance for the evaluation of biochemical toxicity, wide-spectrum antibacterial property and degradation cycle between different tetracyclines. In this study, the white fluorescent Eu/Tb@CDs was synthesized and applied successfully to the identification and detection of the most widely used tetracycline antibiotics (tetracycline (TC), oxytetracycline (OC), chlortetracycline (CC) and doxycycline (DC)) with detection limits all below 1 nM. For the actual water samples with coexistence of the above 4 tetracyclines, their simultaneous morphology identification and accurate quantitative detection can also be realized through simple spectrometric measurement. In addition, the selective and competitive experiments have been carried out on the pollutants widely present in water, and the results have also confirmed that other pollutants could not interfere with the detection of the above 4 tetracyclines. It is undeniable that this work will conveniently and visually reveal the existence information and geographical distribution characteristics of different tetracycline antibiotics in the environment and their action mechanism on organisms., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

A novel multicolor fluorescent nano-probe based on the hybridization of Tb 3+ ion with gold nanoclusters (Au NCs) was synthesized to monitor and on-site visual assay of 2,6-pyridinedicarboxylic acid (DPA), a biomarker of bacterial spores. DPA can replace the water molecule in the center of Tb 3+ and strongly coordinate with Tb 3+ based on the analyte-triggered antenna effect. Simultaneously, the red fluorescence of Au NCs is not influenced after addition of DPA and can be used as steady inside fluorescence reference channel to measure background noise. On this basis, the multicolor fluorescence nano-probe based on Tb 3+ -doped Au NCs for fast analysis of DPA was fabricated. The linear range of this method is 0 to 12.5 μM and the limit of detection is 3.4 nM, which is well below the quantity of DPA concentration of 60 μM released by the spore transmission dose of anthrax infection. The proposed multicolor fluorescence nano-probe was successfully detecting DPA in actual sample with good sensitivity and specificity. In addition, the visual paper-based nano-probe is designed to detect DPA by using the color scanning application of smart phone. This developed platform possesses abroad application prospects with advantages of effective, convenient carrying, simple operation, good selectivity and repeatability., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Two-dimensional difference gel electrophoresis (2D-DIGE) appears to be especially useful in quantitative approaches, allowing the co-separation of proteins of control samples and proteins of treated/disease samples on the same gel, eliminating gel-to-gel variability. The principle of 2D-DIGE is to label proteins prior to isoelectric focusing and use three spectrally resolvable fluorescent dyes, allowing the independent labeling of control and experimental samples. This procedure makes it possible to reduce the number of gels in an experiment, allowing the accurate and reproducible quantification of multiple samples. 2D-DIGE has been found to be an excellent methodical tool in several areas of fish research, including environmental pollution and toxicology, the mechanisms of development and disorders, reproduction, nutrition, evolution, and ecology., (© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

Two-dimensional comparative fluorescence gel electrophoresis (CoFGE) uses an internal standard to increase the reproducibility of coordinate assignment for protein spots visualized on 2D polyacrylamide gels. This is particularly important for samples that need to be compared without the availability of replicates and thus cannot be studied using differential gel electrophoresis (DIGE). CoFGE corrects for gel-to-gel variability by co-running with the sample proteome a standardized marker grid of 80-100 nodes, which is formed by a set of purified proteins. Differentiating of reference and analyte is possible by the use of two fluorescent dyes. Variations in the y-dimension (molecular weight) are corrected by the marker grid. For the optional control of the x-dimension (pI), azo-dyes can be used. Experiments are possible in both vertical and horizontal (h) electrophoresis devices, but hCoFGE is much easier to perform. The CoFGE experimental principle can additionally be used for protein quantification. For data analysis, commercial software has been adapted., (© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

A fused heterocyclic ESIPT imidazo[1,2-a]pyridine-based probes for colorimetric and fluorometric detection of nerve agents simulant sarin (DCP) and tabun (DCNP) are reported. The probes (5b, 6a & 6b) were found to be highly sensitive and selective for the detection of DCNP and DCP at a micromolar concentration within seconds with no observed interference from other various types of analytes. The LOD for 6b towards DCP was found to be 0.6 µM with a linear range from 0 to 8 µM. The low-cost portable cellulose paper strip fabricated with probe 6b for real-time detection of DCP in the gas phase and spiked water has been developed. The paper strip product was found effective in detecting the presence of DCP in water and vapor state with substantial color changes which could be easily observed by the naked eye and under a handheld UV lamp at a wavelength of 365 nm., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

The primary seafood-borne pathogen Vibrio parahaemolyticus seriously threats the health of consumers preferring raw-fish products, becoming a global concern in food safety. In the present study, we found ferrous sulfate (FeSO 4 ), a nutritional iron supplement, could efficiently induce the death of V. parahaemolyticus. Further, the bactericidal mechanisms of FeSO 4 were explored. With a fluorescent probe of Fe 2+ , a significant influx of Fe 2+ was determined in V. parahaemolyticus exposed to FeSO 4 , and the addition of an intracellular Fe 2+ chelator was able to block the cell death. This suggested that cell death in V. parahaemolyticus induced by FeSO 4 was dependent on the influx of Fe 2+ . It was intriguing that we did not observe the eruption of reactive oxygen species (ROS) and lipid hydroperoxides by Fe 2+ , but the application of liproxstatin-1 (a ferroptosis inhibitor) significantly modified the occurrence of cell death in V. parahaemolyticus. These results suggested FeSO 4 -induced cell death in V. parahaemolyticus be a ferroptosis differing from that in mammalian cells. Through transcriptome analysis, it was discovered that the exposure of FeSO 4 disturbed considerable amounts of gene expression in V. parahaemolyticus including those involved in protein metabolism, amide biosynthesis, two-component system, amino acid degradation, carbon metabolism, citrate cycle, pyruvate metabolism, oxidative phosphorylation, and so on. These data suggested that FeSO 4 was a pleiotropic antimicrobial agent against V. parahaemolyticus. Notably, FeSO 4 was able to eliminate V. parahaemolyticus in salmon sashimi as well, without affecting the color, texture, shearing force, and sensory characteristics of salmon sashimi. Taken together, our results deciphered a unique ferroptosis in V. parahaemolyticus by FeSO 4 , and highlighted its potential in raw-fish products to control V. parahaemolyticus., Competing Interests: Declaration of competing interest The authors (Yanyan Zhao, Xiaofeng Kang, Wei Zhou, Jintae Lee，Shuyan Wang, Zhenkun Cui, Hao Zhang, Haizhen Mo, Liangbin Hu) declare that there are no financial interests or personal relationships that will influence the work reported in this paper., (Copyright © 2022. Published by Elsevier B.V.)

A ratiometric fluorescence strategy was proposed based on carbon dots (CDs) and self-assembled copper nanoclusters (CuNCs) driven by Al 3+ ions for S 2- detection. Si-CDs/CuNCs@Al 3+ exhibits blue and red emission under single excitation. Interestingly, the red emission of the CuNCs was regularly quenched while the blue fluorescence emission of the CDs was preserved after continuous addition of S 2- . The fluorescence spectrometer-based S 2- linear range is from 0.5 to 40 μM, with a low detection limit (LOD) of 0.16 μM. The fluorescence response of Si-CDs/CuNCs@Al 3+ to S 2- exhibits a distinct color change process (red to pink to blue), implying feasibility of visual analysis. A portable fluorescence sensing platform was established using the color-to-value conversion function of a smartphone for accurate visualization and quantitative identification of S 2- without spectrometer. The fluorescent test strips prepared with Si-CDs/CuNCs@Al 3+ can conduct on-site visual analysis of S 2- in the water environment more conveniently and quickly. The linear range of S 2- detection based on the smartphone-integrated test strip sensing platform is 1-40 μM, and the LOD is 0.42 μM. This work provides a new horizon for target on-site analysis in environmental samples., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022. Published by Elsevier B.V.)

Staining microplastics (MPs) for fluorescence detection has been widely applied in MP analyses. However, there is a lack of standardized staining procedures and conditions, with different researchers using different dye concentrations, solvents, incubation times, and staining temperatures. Moreover, with the limited types and morphologies of commercially available MPs, a simple and optimized approach to making fluorescent MPs is needed. In this study, 4 different textile dyes, along with Nile red dye for comparison, are used to stain 17 different polymers under various conditions to optimize the staining procedure. The MPs included both virgin and naturally weathered polymers with different sizes and shapes (e.g., fragments, fibers, foams, pellets, beads). We show that the strongest fluorescence intensity occurred with aqueous staining at 70 °C for 3 h with a dye concentration of 5 mg/mL, 55 mg/mL, and 2 µg/mL for iDye dyes, Rit dyes, and Nile red, respectively. Red fluorescent signals are stronger and thus preferred over green ones. The staining procedure did not significantly alter the surface, mass, and chemical characteristics of the particles, based on FTIR and stereomicroscopy. Stained MPs were spiked into freshwater, saltwater, a sediment slurry, and wastewater-activated sludge; even after several days, the recovered particles are still strongly fluoresced. The approach described herein for producing customized fluorescent MPs and quantifying MPs in laboratory-controlled experiments is both straightforward and simple.

A novel portable low-cost Arduino-controlled photo- and fluorimeter for on-site measurements has been developed. The device uses LEDs as a light source and a phototransistor as a light sensor. The circuit is based on the discharge of a capacitor with the photocurrent from the phototransistor. Validation experiments for absorbance measurements were performed by measuring protein concentration using the Bradford method and measuring phosphate ions in water using a commercial test kit. The emission light of the excited fluorescent dyes rhodamine 6G and calcofluor white was measured to validate the usability of the device as a fluorescence photometer. In all validation experiments, similar correlation coefficients and limit of detection could be achieved with the portable photo- and fluorimeter and a laboratory spectrometer and fluorimeter. Real sample analysis was performed, measuring phosphate concentration in freshwater and concentration of green fluorescent protein, extracted from Escherichia coli.

γ-Glutamyl transpeptidase (GGT) is one of the biomarker of cancer, hepatitis, and numerous other diseases. The accurate analysis of GGT is useful for the early diagnosis of these diseases. In this work, Probe 1, a ratiometric fluorescent probe based on 2,3,5,6-tetrafluoroterephthalonitrile, was designed for GGT detection. The results indicated that Probe 1 can sensitively and selectively detect GGT in phosphate buffered solution and complex biological systems (e.g., blood serum). Furthermore, Probe 1 has been successfully applied for ratiometric imaging of GGT in cancer cells and normal cells., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Noble metal nanoclusters have attracted much attention due to their excellent optical properties. In the present work, a silver-doped gold-based bimetallic nanoclusters (Au/Ag NCs) were reasonably designed and prepared through a one-pot method by using 5-mercapto-1-tetrazolea-acetic acid sodium salt (MTAS) as a protector and capping agent. In comparison with the monometallic nanoclusters, Ag-doped metallic nanoclusters show better performance. The particle size of the MTAS-Au/Ag NCs is slightly larger than that of the undoped Au NCs by about 1.86 ± 0.5 nm, and the MTAS-Au/Ag NCs exhibit an emission peak at 635 nm with a quantum yield (QY) of 3.05%. The presence of cysteine (Cys) induces efficient quenching of the photoluminescence of the obtained Au/Ag NCs, achieving the sensitive detection of Cys with a detection limit of 16 nM. The fluorescence quenching rate of the nano fluorescent probe has a linear relationship with the cysteine concentration. Under the best detection conditions, the linear range for Cys detection with MTAS-Au/Ag NCs as a probe is 0.05-25.0 μM. Moreover, this probe has been successfully applied to the analysis of Cys in milk and apples, and a satisfactory recovery rate has been obtained, indicating the effectiveness and reliability of the sensor system for the detection of actual samples., (Copyright © 2022 Elsevier B.V. All rights reserved.)

The plasma membrane, as a highly complex cell organelle, serves as a crucial platform for a multitude of cellular processes. Its collective biophysical properties are largely determined by the structural diversity of the different lipid species it accommodates. Therefore, a detailed investigation of biophysical properties of the plasma membrane is of utmost importance for a comprehensive understanding of biological processes occurring therein. During the past two decades, several environment-sensitive probes have been developed and become popular tools to investigate membrane properties. Although these probes are assumed to report on membrane order in similar ways, their individual mechanisms remain to be elucidated. In this study, using model membrane systems, we characterized the probes Pro12A, NR12S and NR12A in depth and examined their sensitivity to parameters with potential biological implications, such as the degree of lipid saturation, double bond position and configuration ( cis versus trans ), phospholipid headgroup and cholesterol content. Applying spectral imaging together with atomistic molecular dynamics simulations and time-dependent fluorescent shift analyses, we unravelled individual sensitivities of these probes to different biophysical properties, their distinct localizations and specific relaxation processes in membranes. Overall, Pro12A, NR12S and NR12A serve together as a toolbox with a wide range of applications allowing to select the most appropriate probe for each specific research question.

Deciphering the dynamics of intracellular organelles has gained immense attention due to their subtle control over diverse, complex biological processes such as cellular metabolism, energy homeostasis, and autophagy. In this context, molecular materials, including small-organic fluorescent probes and their supramolecular self-assembled nano-/microarchitectures, have been employed to explore the diverse intracellular biological events. However, only a handful of fluorescent probes and self-assembled emissive structures have been successfully used to track different organelle's movements, circumventing the issues related to water solubility and long-term photostability. Thus, the water-soluble molecular fluorescent probes and the water-dispersible supramolecular self-assemblies have emerged as promising candidates to explore the trafficking of the organelles under diverse physiological conditions. In this review, we have delineated the recent progress of fluorescent probes and their supramolecular self-assemblies for the elucidation of the dynamics of diverse cellular organelles with a special emphasis on lysosomes, lipid droplets, and mitochondria. Recent advancement in fluorescence lifetime and super-resolution microscopy imaging has also been discussed to investigate the dynamics of organelles. In addition, the fabrication of the next-generation molecular to supramolecular self-assembled luminogens for probing the variation of microenvironments during the trafficking process has been outlined.

High-performance liquid chromatography-size exclusion chromatography and excitation-emission matrix (EEM) fluorescence spectroscopy were used to analyze the seasonal variations and potential sources of molecular weight (MW) separated light-absorbing chromophores and fluorophores of water-soluble organic compounds (WSOC) and methanol-soluble organic compounds (MSOC) in PM 2.5 in cold areas of northern China. The results showed that the light-absorbing organics in MSOC had larger weight-average MW (M w ) (3.19 kDa) and number-average MW (M n ) (1.13 kDa) compared with WSOC (M w : 1.41 kDa, M n : 0.692 kDa). The light-absorption of organics showed a trend of winter>spring>autumn>summer and increased on air pollution days. Three fluorescent components including humic-like, protein-like, and terrestrial humic-like components in WSOC were extracted by parallel factor analysis (PARAFAC). Fluorophores in WSOC were dominated by humic-like and terrestrial humic-like components (67.7%). Three fluorescent components extracted from MSOC were low oxidation humic-like, polycyclic aromatic hydrocarbon (PAH)-like, and protein-like components respectively. It is worth noting that compared with WSOC, MSOC may have a higher human health risk due to the presence of PAH-like components. The combination of PARAFAC and self-organizing map had the potential to identify potential sources of fluorophores. It provided a new perspective for comprehensively exploring the characteristics of fluorophores in aerosols. This study provided a reference for further understanding the chemical composition and optical properties of organic aerosols in the cold regions of northern China., Competing Interests: Declaration of competing interest The authors declare no conflicts of interest., (Copyright © 2022. Published by Elsevier B.V.)

EDIL3 is a strong and highly accurate diagnostic marker for breast cancer, meanwhile, EDIL3 overexpressed exosomes are novel biomarkers for the early diagnosis of triple-negative breast cancer (TNBC). Here, we proposed a fluorescent detection method for EDIL3 overexpressed exosomes, which is simple and sensitive. Basically, we utilized a magnetic nanospheres (MNS) based liquid sandwich immunoassay strategy. MNS were modified with CD63 aptamers, which can immunologically bound to the CD63 protein on the surface of exosomes. Alexa Fluor 647 labeled anti-EDIL3 antibodies (Anti-EDIL3/AF647) were used as the fluorescent probes to recognize the EDIL3 on exosomes derived from a TNBC cell line (MDA-MB-231). With the target TNBC exosomes present, sandwich structures containing MNS, exosomes and fluorescent probes were formed. After magnetic purification, optical super resolution imaging of the products was conducted to check the specificity of the assay. In addition, fluorescence signals of the products were detected to quantitatively analyze the EDIL3 overexpressed exosomes. The linear range was found to be 7.78 × 10 1 to 7.78× 10 6 particles μ l -1 . The detection limit was approximately 10 particles μ l -1 . The feasibility of the method for the detection of exosomes in complex biological samples was also demonstrated. Such a simple and sensitive detection method for EDIL3 overexpressed exosomes holds a great potential in clinical diagnosis of TNBC., (© 2022 IOP Publishing Ltd.)

A great number of anions exist in biological systems and natural environment, and are highly relevant to human health and environment quality. It is necessary to develop simple and effective sensors to differentiate and identify those similar or different anions. Here, an imidazolium-modified bispyrene-based fluorescent amphiphilic probe DPyDIM was synthesized and its aggregates were applied to detect and discriminate various anions. The fluorescent aggregates exhibit ratiometric responses to different types of anions. Moreover, the ratiometric responses to different types of anions are featured with multiple-wavelength cross-reactivity. The collection of fluorescence variation at four typical wavelengths can generate distinct recognition patterns to specific anions. The heat map and principal component analysis results verify that this single fluorescent sensor system can effectively and sensitively identify 16 kinds of anions that belong to phosphorus-containing, sulfur-containing anions, and anionic surfactants. The cross-reactive sensing of the amphiphilic fluorescent aggregates was attributed to the different influences on the aggregation behaviors of the probes by different anions. The present work provides a promising strategy for effective detection and discrimination of multiple anions by employing dynamic fluorescent aggregates as a sensing platform.

Real-time quantification of glutathione (GSH) concentrations in living cells is essential for understanding GSH-related pathological events. However, most existing fluorescent probes are not suitable for this purpose due to their irreversible response mechanisms. Only a few probes have been successfully developed to realize this goal to date. Herein, we presented a fluorescent probe (QG-S) for quantifying cellular GSH and monitoring GSH dynamic variations. The probe QG-S was successfully employed in real-time monitoring of the dynamic variations of GSH in living cells upon reversible treatment with a GSH inhibitor (N-methylmaleimide) by using confocal microscopy. Moreover, the probe QG-S showed extremely low cytotoxicity and was facilely used to quantitatively determine the GSH concentration (4.25 mM) in living HeLa cells., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Reactive oxygen species (ROS) are an important component for maintaining normal physiological activities in organisms, and abnormal changes in their level are often accompanied by many diseases. As the two most representative components of ROS, HClO and H 2 O 2 play vital roles in many physiological and pathological processes and are interdependent and mutually transformable. Although there is a lot of work that has specifically detected HClO or H 2 O 2 , there are few reports on the simultaneous differential detection of HClO and H 2 O 2 . Here, we report a ratio-based fluorescent probe capable of simultaneously distinguishing HClO and H 2 O 2 based on making the best use of the untapped potential of coumarin derivatives. This probe was triumphantly put into use in the discriminative identification of HClO and H 2 O 2 in aqueous media with high sensitivity and selectivity, and the probe was appropriate in a wide pH range. Furthermore, the imaging experiment for HClO and H 2 O 2 in cells and zebrafish was eventually proven to be feasible. Importantly, this probe was qualified for monitoring the variation of HClO and H 2 O 2 levels in organisms with alcoholic liver injury.

The fluorescent organic nanoparticles (FONs)-based sensor has been attracting great attention in recent years. There are still big challenges in the preparation and application of FONs-based sensor. In this study, a FONs-based sensor was designed and developed through facile hydrothermal process using 3-perylenecarboxaldehyde (PlCA) as the fluorophore and L-methionine (Met) as the recognition site for mercury ions. According to the experimental results, the fluorescence intensity of the as-prepared PlCA-M would decrease when adding Hg 2+ and the mechanism was extrapolated to be photoinduced electron transfer inducing by specific coordination interaction. The acquired PlCA-M-based sensor was used to monitor Hg 2+ in several real samples (environmental water, tea, and apple) with the limit of detection being 60 nM. Remarkably, a visual detection device based on FONs, SDS-PAAG (sodium dodecyl sulfate polyacrylamide gel) @PlCA-M was firstly constructed and successfully used to Hg 2+ semi-quantitation by naked eyes. In addition, the acquired FONs was applied into imaging tool for security information detection and identified as solid-state luminescent material for the first time., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Hydrogen sulfide (H 2 S) is a well-known toxic gas with the odor of rotten eggs. Several reaction-based electrochemiluminescence (ECL) chemosensors for H 2 S have been developed; however, no homogeneous ECL probe with high selectivity toward H 2 S in aqueous media has been reported. Herein, we report an iridium(III) complex-based ECL chemodosimetric probe employing two 7-nitrobenz-2-oxa-1,3-diazol-4-yl (NBD) groups known as a photo-induced electron transfer quencher and a reaction site for the selective detection of H 2 S; the detection mechanism involves H 2 S being clearly distinguished from biothiols based on the different cleavage rates of the two NBD groups and extremely weak ECL interferences caused by reaction by-products. The probe was rationally designed to improve selectivity toward H 2 S within the ECL analysis platform by enabling the removal of nonspecific background signals observed via fluorescence analysis. This analytical system exhibited remarkable selectivity toward H 2 S, a rapid reaction rate, and high sensitivity (LOD = 57 nM) compared to conventional fluorescence methods. Furthermore, the probe could successfully quantify H 2 S in tap water samples and commercial ammonium sulfide solutions, which demonstrates the effectiveness of this probe in field monitoring.

In this study, the pros and cons of the most relevant L-format devices reported in the literature for measuring steady-state fluorescence polarization/anisotropy are identified. Combining all this information, and with the use of modern elements for the acquisition, treatment, and recording of signals, a modern, simple, and economical L-format accessory is implemented to rapidly and continuously record steady-state fluorescence anisotropy. This device can be adapted to the majority of the commercial spectrofluorometers (or fluorometers). During the measurement, the emission polarizer is in permanent rotation by means of a Gimbal brushless DC motor, and as a result the recorded fluorescence signal is sinusoidal. The maximums and minimums of this signal, which are obtained with the help of LabVIEW tools, allow recording the fluorescence anisotropy. The LabVIEW applications developed for this investigation are freely available, so it is not necessary to have LabVIEW software., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Nowadays, palladium has been widely used in many fields, which facilitates all aspects of our life. However, it may cause water and soil pollution and bring irreversible damage to the environment and organisms. Developing a fluorescence probe for rapid, highly sensitive and selective detection of palladium is still a poser. In this work, we designed and synthesized a novel fluorescence probe (RHS) for specific detection of palladium. Based on Pd 0 -mediated Tsuji-Trost reaction, the fluorescence probe was constructed by a rhodol derivative as thefluorophore and an allyl carbonate moiety as the specific palladium reactive site. The probe displayed excellent properties for detecting palladium, such as high selectivity and sensitivity, rapid response (20 min) and large Stokes shift (155 nm). The detection limit was determined to be as low as 0.140 μM with a linear range from 20 to 80 μM. After addition of palladium in RHS solution, the color of the solution turned from yellow to blue, indicating palladium could be monitored by the naked eyes. Moreover, probe RHS was successfully applied to palladium detection in environmental water samples. Importantly, with low cytotoxicity and good biocompatibility, the probe could monitor palladium in living cells., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Dysregulated energy metabolism has recently been recognized as an emerging hallmark of cancer. Tumor cells, which are characterized by abnormal glycolysis, exhibit a lower extracellular pH (6.5-7.0) than normal tissues (7.2-7.4), providing a promising target for tumor-specific imaging and therapy. However, most pH-sensitive materials are unable to distinguish such a subtle pH difference owing to their wide and continuous pH-responsive range. In this study, we developed an efficient strategy for the fabrication of a tumor metabolic acidity-activatable calcium phosphate (CaP) fluorescent probe (termed MACaP9). Unlike traditional CaP-based biomedical nanomaterials, which only work within more acidic organelles, such as endosomes and lysosomes (pH 4.0-6.0), MACaP9 could not only specifically respond to the tumor extra-cellular pH but also rapidly convert pH variations into a distinct fluorescence signal to visually distinguish tumor from normal tissues. The superior sensitivity and specificity of MACaP9 enabled high-contrast visualization of a broad range of tumors, as well as small tumor lesions., (Copyright © 2021 Science China Press. Published by Elsevier B.V. All rights reserved.)

Measurement of the nitric oxide (NO) concentration in living cells in the physiological nanomolar range is crucial in understanding NO biochemical functions, as well as in characterizing the efficiency and kinetics of NO delivery by NO-releasing drugs. Here, we show that fluorescence correlation spectroscopy (FCS) is perfectly suited for these purposes, due to its sensitivity, selectivity, and spatial resolution. Using the fluorescent indicators, diaminofluoresceins (DAFs), and FCS, we measured the NO concentrations in NO-producing living human primary endothelial cells, as well as NO delivery kinetics, by an external NO donor to the immortal human epithelial living cells. Due to the high spatial resolution of FCS, the NO concentration in different parts of the cells were also measured. The detection of nitric oxide by means of diaminofluoresceins is much more efficient and faster in living cells than in PBS solutions, even though the conversion to the fluorescent form is a multi-step reaction.

Fluorescence lifetime imaging microscopy (FLIM) is a powerful tool to quantify molecular compositions and study molecular states in complex cellular environment as the lifetime readings are not biased by fluorophore concentration or excitation power. However, the current methods to generate FLIM images are either computationally intensive or unreliable when the number of photons acquired at each pixel is low. Here we introduce a new deep learning-based method termed flimGANE (fluorescence lifetime imaging based on Generative Adversarial Network Estimation) that can rapidly generate accurate and high-quality FLIM images even in the photon-starved conditions. We demonstrated our model is up to 2,800 times faster than the gold standard time-domain maximum likelihood estimation (TD&#95;MLE) and that flimGANE provides a more accurate analysis of low-photon-count histograms in barcode identification, cellular structure visualization, Förster resonance energy transfer characterization, and metabolic state analysis in live cells. With its advantages in speed and reliability, flimGANE is particularly useful in fundamental biological research and clinical applications, where high-speed analysis is critical., (© 2022. The Author(s).)

Organic near-infrared room temperature phosphorescence materials have unparalleled advantages in bioimaging due to their excellent penetrability. However, limited by the energy gap law, the near-infrared phosphorescence materials (>650 nm) are very rare, moreover, the phosphorescence lifetimes of these materials are very short. In this work, we have obtained organic room temperature phosphorescence materials with long wavelengths (600/657-681/732 nm) and long lifetimes (102-324 ms) for the first time through the guest-host doped strategy. The guest molecule has sufficient conjugation to reduce the lowest triplet energy level and the host assists the guest in exciton transfer and inhibits the non-radiative transition of guest excitons. These materials exhibit good tissue penetration in bioimaging. Thanks to the characteristic of long lifetime and long wavelength emissive phosphorescence materials, the tumor imaging in living mice with a signal to background ratio value as high as 43 is successfully realized. This work provides a practical solution for the construction of organic phosphorescence materials with both long wavelengths and long lifetimes., (© 2022. The Author(s).)

Optosensorics is the direction of research possessing the possibility of non-invasive monitoring of the concentration of intracellular ions or activity of intracellular components using specific biosensors. In recent years, genetically encoded proteins have been used as effective optosensory means. These probes possess fluorophore groups capable of changing fluorescence when interacting with certain ions or molecules. For monitoring of intracellular concentrations of chloride ([Cl - ] i ) and hydrogen ([H + ] i ) the construct, called ClopHensor, which consists of a H + - and Cl - -sensitive variant of the enhanced green fluorescent protein (E 2 GFP) fused with a monomeric red fluorescent protein (mDsRed) has been proposed. We recently developed a line of transgenic mice expressing ClopHensor in neurons and obtained the map of its expression in different areas of the brain. The purpose of this study was to examine the effectiveness of transgenic mice expressing ClopHensor for estimation of [H + ] i and [Cl - ] i concentrations in neurons of brain slices. We performed simultaneous monitoring of [H + ] i and [Cl - ] i under different experimental conditions including changing of external concentrations of ions (Ca 2+ , Cl - , K + , Na + ) and synaptic stimulation of Shaffer's collaterals of hippocampal slices. The results obtained illuminate different pathways of regulation of Cl - and pH equilibrium in neurons and demonstrate that transgenic mice expressing ClopHensor represent a reliable tool for non-invasive simultaneous monitoring of intracellular Cl - and pH.

Fluorescent whitening agents (FWAs) are very important chemical additives that are widely applied in the industrial production field. The history of global FWA production and use spans more than 60 years, but the environmental fate of FWAs has been less reported in the public literature and most studies predate 2000; in addition, the studied FWAs were still limited to FWA71 and FWA351. In this study, the occurrence and distribution of 9 commonly used FWAs in a lake in North China were reported for the first time. We found that 6 target FWAs were prevalent in the lake, and the concentration levels were usually at the ng L -1 level. Decreasing FWA levels with increasing distance from the estuary area were observed in summer. FWA135, FWA185, and FWA367, the most detected 3 FWAs, with the ecological risk at high levels, and ΣRQ >10 were obtained from all the investigated samples, suggesting that all the sampling sites could be considered with certain ecological risk for aquatic life. As a category of heavily and widely used dyes, FWAs in environmental media have been ignored for a long time. Substantial additional research needs to be conducted to determine the environmental behavior and ecological toxicology of FWAs., (Copyright © 2021. Published by Elsevier Ltd.)

In living organisms, redox reactions play a crucial role in the progression of disorders accompanied by the overproduction of reactive oxygen and reactive chlorine species, such as hydrogen peroxide and hypochlorous acid, respectively. We demonstrate that green fluorescence graphene quantum dots (GQDs) can be employed for revealing the presence of the hypochlorous acid in aqueous solutions and cellular systems. Hypochlorous acid modifies the oxygen-containing groups of the GQD, predominantly opens epoxide ring C-O-C, forms excessive C=O bonds and damages the carbonic core of GQDs. These changes, which depend on the concentration of the hypochlorous acid and exposure time, manifest themselves in the absorbance and fluorescence spectra of the GQD, and in the fluorescence lifetime. We also show that the GQD fluorescence is not affected by hydrogen peroxide. This finding makes GQDs a promising sensing agent for selective detecting reactive chlorine species produced by neutrophils. Neutrophils actively accumulate GQDs allowing to visualize cells and to examine the redox processes via GQDs fluorescence. At high concentrations GQDs induce neutrophil activation and myeloperoxidase release, leading to the disruption of GQD structure by the produced hypochlorous acid. This makes the GQDs a biodegradable material suitable for various biomedical applications., (© 2021 IOP Publishing Ltd.)

The cyclic signal amplification technology has been widely applied for the ultrasensitive detection of many important biomolecules, such as nucleic acids, proteins, enzymes, adenosine triphosphate (ATP), metal ions, exosome, etc. Due to their low content in the complex biological samples, traditional detection methods are insufficient to satisfy the requirements for monitoring those biomolecules. Therefore, effective and sensitive biosensors based on cyclic signal amplification technology are of great significance for the quick and simple diagnosis and treatment of diseases. Fluorescent biosensor based on cyclic signal amplification technology has become a research hotspot due to its simple operation, low cost, short time, high sensitivity and high specificity. This paper introduces several cyclic amplification methods, such as rolling circle amplification (RCA), strand displacement reactions (SDR) and enzyme-assisted amplification (EAA), and summarizes the research progress of using this technology in the detection of different biomolecules in recent years, in order to provide help for the research of more efficient and sensitive detection methods., (© 2021. The Author(s).)