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Industrialization has led to environmental pollution with various hazardous chemicals including pollution with metals. In this regard, the development of highly efficient analytical methods for their determination has received considerable attention to ensure public safety. Currently, scientists are paying more and more attention to the automation of analytical methods, since it permits fast, accurate, and sensitive analysis with minimal exposure of analysts to hazardous substances. This review discusses the automated methods with fluorescent detection developed for metal determination since 2000. It is evident that flow-injection analysis (FIA) with no preconcentration or with solid-phase preconcentration are predominant compared to liquid-phase preconcentration systems. FIA systems are also more widespread than sequential injection analysis (SIA) systems. Moreover, a significant number of works have been devoted to chromatography-based methods. Atomic fluorescence detectors significantly prevail over molecular fluorescence detectors. It must be highlighted that most of the methods result in good figures of merit and performance characteristics, demonstrating their superiority in comparison with manual systems.

Setophoma terrestris is an important phytopathogenic fungus listed by China as a harmful fungus subject to phytosanitary import control. This pathogen is a threat to a wide range of plants, particularly as the causal agent of onion pink root rot, one of the most severe diseases of onions. In order to provide rapid identification and early warning of S. terrestris and prevent its spread, we have developed a rapid, accurate, and visually intuitive diagnostic assay for this pathogen, by utilizing recombinase polymerase amplification (RPA), coupled with CRISPR/Cas12a cleavage and fluorescence-based detection systems or paper-based lateral flow strips. The developed RPA-CRISPR/Cas12a assay exhibited remarkable specificity for the detection of S. terrestris. Moreover, this protocol can detect the pathogen at a sensitivity level of 0.01 pg/μL, which significantly outperforms the 1 pg/μL sensitivity achieved by the existing qPCR-based detection method. The entire diagnostic procedure, including DNA extraction, the RPA reaction, the Cas12a cleavage, and the result interpretation, can be accomplished in 40 min. Furthermore, the successful application of the assay in infected plant samples highlighted its potential for rapid and accurate pathogen detection in agricultural settings. In summary, this RPA-CRISPR/Cas12a diagnostic method offers a potentially valuable technological solution for quarantine and disease management.

Antioxidants are substances that have the ability to resist or delay oxidative damage. Antioxidants can be used not only for the diagnosis and prevention of vascular diseases, but also for food preservation and industrial production. However, due to the excessive use of antioxidants, it can cause environmental pollution and endanger human health. It can be seen that the development of antioxidant detection technology is important for environment/health maintenance. It is found that traditional detection methods, including high performance liquid chromatography, gas chromatography, etc., have shortcomings such as cumbersome operation and high cost. In contrast, the nanozyme-based detection method features advantages of low cost, simple operation, and rapidity, which has been widely used in the detection of various substances such as glucose and antioxidants. This article focuses on the latest research progress of nanozymes for antioxidant detection. Nanozymes for antioxidant detection are classified according to enzyme-like types. Different types of nanozyme-based sensing strategies and detection devices are summarized. Based on the summary and analysis, one can find that the development of commercial nanozyme-based devices for the practical detection of antioxidants is still challenging. Some emerging technologies (such as artificial intelligence) should be fully utilized to improve the detection sensitivity and accuracy. This article aims to emphasize the application prospects of nanozymes in antioxidant detection and to provide new ideas and inspiration for the development of detection methods.

Fluorescence flow cytometry is a powerful instrument to distinguish cells or particles labelled with high-specificity fluorophores. However, traditional flow cytometry is complex, bulky, and inconvenient for users to adjust fluorescence channels. In this paper, we present a modular fluorescence flow cytometry (M-FCM) system in which fluorescence channels can be flexibly arranged. Modules for particle focusing and fluorescence detection were developed. After hydrodynamical focusing, the cells were measured in the detection modules, which were integrated with in situ illumination and fluorescence detection. The signal-to-noise ratio of the detection reached to 33.2 dB. The crosstalk among the fluorescence channels was eliminated. The M-FCM system was applied to evaluate cell viability in drug screening, agreeing well with the commercial cytometry. The modular cytometry presents several outstanding features: flexibility in setting fluorescence channels, cost efficiency, compact construction, ease of operation, and the potential to upgrade for multifunctional measurements. The modular cytometry provides a multifunctional platform for various biophysical measurements, e.g., electrical impedance and refractive-index detection. The proposed work paves an innovative avenue for the multivariate analysis of cellular characteristics.

A new green and highly sensitive method for the determination of rhodamine B (RhB) by deep eutectic solvent-based vortex-assisted liquid–liquid microextraction with fluorescence detection (DES-VALLME-FLD) was developed. The extraction efficiency of conventional solvents and different deep eutectic solvent (DES) systems composed of tetrabutylammonium bromide (TBAB) and an alcohol (hexanol, octanol, or decanol) in different ratios were compared. DFT calculations of intermolecular electrostatic and non-covalent interactions of the most stable RhB forms with DES and water explain the experimental DESs’ extraction efficiency. Semiempirical PM7 computations were used to obtain Hansen solubility parameters, which supported the good solubility of the monocationic RhB form in selected DESs. The dependence of the linear calibration of microextraction into 100 µL DES was observed in the RhB calibration range from 0.2 to 10.0 µg L−1 with a correlation coefficient of R2 = 0.9991. The LOD value was calculated to be 0.023 µg L−1. The accuracy and precision of the proposed method were verified over two days with RSD values of 2.9 to 4.1% and recovery of 94.6 to 103.7%. The developed method was applied to the determination of RhB in real samples (tap water, energy drink, and lipstick).

Staphylococcus aureus exists widely in the natural environment and is one of the main food-borne pathogenic microorganisms causing human bacteremia. For safe food management, a rapid, high-specificity, sensitive method for the detection of S. aureus should be developed. In this study, a platform for detecting S. aureus (nuc gene) based on isothermal amplification (loop-mediated isothermal amplification—LAMP, recombinase polymerase amplification—RPA) and the clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas12a) proteins system (LAMP, RPA-CRISPR/Cas12a) was proposed. In this study, the LAMP, RPA-CRISPR/Cas12a detection platform and immunochromatographic test strip (ICS) were combined to achieve a low-cost, simple and visualized detection of S. aureus. The limit of visual detection was 57.8 fg/µL of nuc DNA and 6.7 × 102 CFU/mL of bacteria. Moreover, the platform could be combined with fluorescence detection, namely LAMP, RPA-CRISPR/Cas12a-flu, to establish a rapid and highly sensitive method for the detection of S. aureus. The limit of fluorescence detection was 5.78 fg/µL of genomic DNA and 67 CFU/mL of S. aureus. In addition, this detection platform can detect S. aureus in dairy products, and the detection time was ~40 min. Consequently, the isothermal amplification CRISPR/Cas12a platform is a useful tool for the rapid and sensitive detection of S. aureus in food.

Digital polymerase chain reaction (PCR) technology in microfluidic systems often results in bubble formation post-amplification, leading to microdroplet fragmentation and compromised detection accuracy. To solve this issue, this study introduces a method based on the constant pressure regulation of microdroplets during PCR within microfluidic chips. An ideal pressure reference value for continuous pressure control was produced by examining air solubility in water at various pressures and temperatures as well as modeling air saturation solubility against pressure for various temperature scenarios. Employing a high-efficiency constant pressure device facilitates precise modulation of the microfluidic chip’s inlet and outlet pressure. This ensures that air solubility remains unsaturated during PCR amplification, preventing bubble precipitation and maintaining microdroplet integrity. The device and chip were subsequently utilized for quantitative analysis of the human epidermal growth factor receptor (EGFR) exon 18 gene, with results indicating a strong linear relationship between detection signal and DNA concentration within a range of 101–105 copies/μL (R2 = 0.999). By thwarting bubble generation during PCR process, the constant pressure methodology enhances microdroplet stability and PCR efficiency, underscoring its significant potential for nucleic acid quantification and trace detection.

Adenosine is a vital biological small molecule that regulates various physiological processes in the human body. A high expression of adenosine in cells can facilitate tumor growth. Therefore, detecting adenosine is crucial for early disease diagnosis. In this paper, we designed a fluorescent biosensor for the sensitive detection of adenosine based on the cationic comb-type copolymer PLL-g-Dex for assisted rapid hybridization of nucleic acids at room temperature. In this strategy, adenosine preferentially binds to the aptamer immobilized on the surface of magnetic nanobeads, releasing free aDNA in solution as the primer strand, which rapidly forms DNA nanowires with auxiliary probes of bDNA with the assistance of PLL-g-Dex. SYBR Green I is embedded in DNA duplexes to generate strong fluorescence. The experimental results showed that PLL-g-Dex promotes DNA hybridization reactions at room temperature to form ultra-long DNA nanowires, thus achieving signal amplification and shortening the detection time. In addition, magnetic nanobeads can reduce the background signal during the reaction. Compared with several previous studies on the fluorescence detection of adenosine, this strategy has a lower detection limit of 2.32 nM. Furthermore, this novel system exhibited a good detection performance even under complex environments, such as serum, providing some reference for the quantitative detection of adenosine in early disease diagnosis.

High-Performance Liquid Chromatography (HPLC) combined with ultraviolet/visible (UV/Vis) or diode array detection (DAD) is routinely used for drug quantification in R&D around the world. However, it may lack the sensitivity required for bioanalytical studies. On the other hand, HPLC with fluorescence detection (FLD) is a cost-effective alternative that significantly increases drug signals, enabling the detection of compounds at very low concentrations. Pitavastatin is a lipid-lowering drug that contains the structure of quinoline, a highly fluorescent molecule. Recently, it has gained interest due to its pleiotropic effects on different conditions. Bearing this in mind, an HPLC-FLD method was developed and validated for the quantification of pitavastatin in human plasma. Overall, a signal gain of 54–70 times compared to that of UV detection was achieved when using fluorescence. Sample preparation included one-step protein precipitation with acetonitrile, followed by centrifugation and filtration prior to injection. Pitavastatin was separated from endogenous matrix interferents using a C18 column and by applying gradient elution. Atorvastatin was used as an internal standard. Accordingly, the method was shown to be selective, specific, and sensitive, with a lower limit of quantification of 3 ng/mL and complete absolute and relative recoveries higher than 94%. The method was linear over the wide concentration range of 3–900 ng/mL (R2 = 0.998), accurate (bias < 7.15%), and precise (RSD < 9.63%). This method allows for the therapeutic monitoring of patients treated with pitavastatin but can also support novel clinical studies of this drug in human plasma.

The response performances of the crystalline organic fluorescence probe are highly dependent on the long-range ordered arrangement of crystalline structure. Herein, a novel organic crystalline fluorescent probe with a high quantum yield was established through the rapid self-assembly of 1,2,4,5-Tetrakis (4-carboxyphenyl) benzene (H4TCPB) and DMF molecules. Each H4TCPB, which connects to four DMF molecules through hydrogen bonds, acts as the structural unit. The building units are packed by π–π, lone pair···π, and lone pair···lone pair interactions to form solid-state crystalline materials. H4TCPB·4DMF exhibits distinct blue fluorescent under UV light, while the quantum yield is as high as 89.02% and the fluorescence lifetime is 1.95 ns. The H4TCPB·4DMF nanocrystal exhibits a specific fluorescence quench sensibility to tetracycline (TC), compared with the common chemicals and ions in environmental water. Moreover, the test results can be obtained quickly and are easily visible to the naked eye. The limit of detection for TC is as low as 12 nM in an aqueous solution. Spectral analysis and density functional theory (DFT) theoretical calculations were used to explain the fluorescence quenching mechanism of H4TCPB·4DMF toward TC, which could be attributed to the photoinduced electron transfer occurring from H4TCPB·4DMF to TC. Our work enriches the database of crystalline luminescent materials and provides theoretical support for the design and mechanical studies of organic fluorescent probes.

Apomixis is a unique reproductive process that produces fertile offspring without the combination of sperm and egg cells. This process perfectly reproduces maternal DNA, making it possible to fix heterosis during reproduction. Malus shizongensis is a newly discovered species that is closely related to Malus hupehensis Rehd. After de-male bagging, it was found that the fruit set rate reached 78.7%. Preliminary analysis indicated that M. shizongensis have apomictic reproductive characteristics. In this work, we employed paraffin sectioning and electron scanning microscopy to explore apomixis in M. shizongensis during the development of male–female gametes and embryo sacs. Stigma fluorescence assays showed that pollen germination was normal, but less pollen entered the ovaries. Additionally, analysis of anthers indicated the presence of dysplasia and paraffin sectioning revealed that the pollen mother cells were aborted due to abnormal disintegration of the tapetum layer. Taken together, our results indicate that the primary causes of apomixis in M. shizongensis are anther dysplasia and male gamete development failure, resulting in reduced pollen tube entry into ovaries and reduced reproduction of female gametes. In conclusion, this study provide a theoretical basis and technical supports for apple stock breeding and apple industry development.

Octreotide is a synthetic cyclic octapeptide analogue of somatostatin-14. It is mainly administered for the treatment of acromegaly, severe diarrhea, and neuroendocrine neoplasias. In this work, a hydrophilic interaction liquid chromatography (HILIC) method with fluorescence (FL) detection was developed and validated for the quantitation of octreotide in solutions for injection. Chromatographic separation was performed on an XBridge®-HILIC analytical column under isocratic elution with a short chromatographic run time of less than 10 min. The mobile phase consisted of ammonium bicarbonate 8.6 mM (pH 8.1)/acetonitrile 35/65 (v/v). The high sensitivity and selectivity of the fluorescence detection, with the excitation wavelength (λexcitation) set at 280 nm and the emission wavelength set at (λemission) 330 nm, enabled a simple sample preparation procedure that included only dilution steps. The calibration curve showed good linearity with a correlation coefficient greater than 0.998. The method was successfully applied to the analysis of commercially available octreotide injection forms.

A device and method for the constant pressure regulation of microdroplet PCR in microfluidic chips are developed to optimize for the microdroplet movement, fragmentation, and bubble generation in microfluidic chips. In the developed device, an air source device is adopted to regulate the pressure in the chip, such that microdroplet generation and PCR amplification without bubbles can be achieved. In 3 min, the sample in 20 μL will be distributed into nearly 50,000 water-in-oil droplets exhibiting a diameter of about 87 μm, and the microdroplet will be subjected to a close arrangement in the chip without air bubbles. The device and chip are adopted to quantitatively detect human genes. As indicated by the experimental results, a good linear relationship exists between the detection signal and DNA concentration ranging from 101 to 105 copies/μL (R2 = 0.999). The microdroplet PCR devices based on constant pressure regulation chips exhibit a wide variety of advantages (e.g., achieving high pollution resistance, microdroplet fragmentation and integration avoidance, reducing human interference, and standardizing results). Thus, microdroplet PCR devices based on constant pressure regulation chips have promising applications for nucleic acid quantification.

Eco-friendly liquid chromatographic methods for measuring ergotamine (EGT) are scant in the published database. Accordingly, the goal of the current study was to develop a high-performance thin-layer chromatography (HPTLC) method for fluorescence detection of EGT in commercially available tablets. This approach was based on the application of ethyl alcohol–water (80:20 v/v) as the eco-friendly eluent mixture. The fluorescence detection of EGT was carried out at 322 nm. The greenness score of the present approach was evaluated by “Analytical GREENness (AGREE)” technology. The present approach for measuring EGT in the 25–1000 ng band−1 range was linear. The present assay for fluorescence detection of EGT was validated successfully by ICH guidelines for various parameters. The method was found to be rapid, sensitive, eco-friendly, and stability-indicating. The computed AGREE index for the current strategy was 0.84, displaying outstanding greenness features. The present methodology successfully separated the EGT degradation products under forced-degradation circumstances, exhibiting its stability-indicating qualities and selectivity. An amount of 99.33% of EGT was found in commercial formulations, indicating the validity of the current method for pharmaceutical analysis of EGT in commercial products. The results showed that EGT in commercial products might be regularly measured by the existing method.

Barley (Hordeum vulgare) is one of the most widely cultivated crops for feedstock and beer production, whereas lupins (Lupinus spp.) are grown as fodder and their seeds are a source of protein. Both species produce the allelopathic alkaloids gramine and hordenine. These plant-specialized metabolites may be of economic interest for crop protection, depending on their tissue distribution. However, in high concentrations they pose a health risk to humans and animals that feed on them. This study was carried out to develop and validate a new method for monitoring these alkaloids and their related metabolites using fluorescence detection. Separation was performed on an HSS T3 column using slightly acidified water-acetonitrile eluents. Calibration plots expressed linearity over the range 0.09–100 pmol/µL for gramine. The accuracy and precision ranged from 97.8 to 123.4%,

We developed an extensible LED-induced fluorescence detection module with a highly integrated and ultra-compact structure. A target-oriented design methodology was used to demonstrate the module’s optimal design. Lucigenin solution was used as a test sample in evaluation trials to demonstrate the module’s quantitative fluorescence detection capability. Results showed that the integrated module has an outstanding linear response in the range of 0–1 μmol·L−1, with sensitivity and limit of detection (LOD) of 0.1692 V/μmol·L−1 and 0.03 μmol·L−1, respectively. Statistical analyses showed that our integrated module has extremely high repeatability and accuracy, i.e., the values of Pearson’s correlation coefficient and root-mean-square error exceeded 0.9995 and 1.8‰, respectively. More importantly, the integrated module possesses favorable extensibility and can realize on-demand rapid fluorescence-signal detection of other targets using appropriate parameter combinations. This module offers new opportunities for reliable, cost-effective and easy-to-use fluorescence-signal detection, especially in resource-constrained fluorescence detection applications.

Continuous-flow PCR (CF-PCR) can realize rapid DNA amplification because of the high temperature variation rate. However, off-line detection methods for PCR may induce cross contamination. To overcome this problem, we herein fabricated an integrated CF-PCR and electrophoresis microfluidic chip. The optimal voltage applied in the electrophoresis part of the microfluidic chip was achieved by simulation in COMSOL. Coating the inside wall of the microchannel can inhibit electroosmotic flow and improve the resolution for DNA fragments. The temperature distribution of the serpentine part can meet the PCR and has no obvious suppressive effect on sample separation. Finally, we have performed the amplification of target genes for Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola and detected the corresponding PCR products in the microfluidic chip within 11 min. Such work provides a new method for the rapid detection of bacteria.

In this study, three compounds A1, A2, and A3 and fluorescent probes T1, T2, T3, and T4 were designed and synthesized. 1H NMR, 13C NMR, and MS characterization and elemental analysis were used to confirm A1–A3 and T1–T4. A1–A3 and T1–T4 formed diagnostic molecules by “click” reactions. A1–A3 and T1–T4 did not significantly increase cell death at concentrations of 80 μmol/L. Preliminary screening of the compounds for antibacterial activity revealed that A2 has better antibacterial activity against Agrobacterium tumefaciens. The synthesized compounds and fluorescent probes can be targeted and combined in the physiological condition to form diagnostic molecules for fluorescence detection of Agrobacterium tumefaciens. The binding sites of A1–A3 were deduced theoretically using the AutoDock Vina software docking tool. Further study of the mechanism of the antibacterial action of these compounds is likely to identify new agents against resistant bacterial strains.

A simple, selective, rapid, sensitive and less costly green automated solid phase extraction bio-analytical high-performance liquid chromatographic-based technique with fluorescence detection (Aut-SPE-BA-HPLC-FL) for the quantification of levofloxacin in human serum samples has been developed and validated. The serum samples were loaded into the chromatographic system without prior treatment and then injected into short (20 mm × 4.6 mm, 20 µm) protein-coated (PC) µBondapak CN (µBCN) silica pre-column (PC-µBCN-pre-column). Levofloxacin was retained and pre-concentrated on the head of the PC-µBCN-pre-column, while proteins and other polar components were eliminated using phosphate buffer saline (PBS), pH 7.4, as the first mobile phase in the extraction step. Levofloxacin is then transferred to the analytical column; ZORBAX Eclipse XDB-C18 (150 mm × 46 mm, 5 µm), through the aid of a column-switching valve technique, on-throughs the elution mode using the second mobile phase containing a methanol and phosphate buffer (0.05 M, pH 5) in a ratio of 70:30 (v/v). Levofloxacin signals were detected using a fluorescence detector operated at excitation/emission wavelengths of 295/500 nm. The proposed Aut-SPE-BA-HPLC-FL methodology showed linearity over a levofloxacin concentration range of 10–10,000 ng/mL (r2 = 0.9992), with good recoveries ranging from 87.12 to 97.55%. Because of the validation qualities in terms of linearity, recovery, precision, accuracy, selectivity and robustness, the Aut-SPE-BA-HPLC-FL method has been used in some clinical trials for therapeutic drug monitoring and the pharmacokinetic study of levofloxacin in human serum.

With the development of nanotechnology, fluorescent silicon nanomaterials have been synthesized and applied in various areas. Among them, silicon quantum dots (SiQDs) are a new class of zero-dimensional nanomaterials with outstanding optical properties, benign biocompatibility, and ultra-small size. In recent years, SiQDs have been gradually utilized for constructing high-performance fluorescent sensors for chemical or biological analytes. Herein, we focus on reviewing recent advances in SiQD-based fluorescent biosensors from a broad perspective and discussing possible future trends. First, the representative progress for synthesizing water-soluble SiQDs in the past decade is systematically summarized. Then, the latest achievement of the design and fabrication of SiQD-based fluorescent biosensors is introduced, with a particular focus on analyte-induced photoluminescence (fluorescence) changes, hybrids of SiQDs with other materials or molecules, and biological ligand-modification methods. Finally, the current challenges and prospects of this field are highlighted.

The applicability of capillary electrophoresis (CE) with light-emitting diode-induced fluorescence detection (LEDIF) for the separation of sugars in honey samples was studied. An amount of 25 mM ammonium acetate (pH 4.5) with 0.3% polyethylene oxide (PEO) was found to be optimal for the efficient separation of carbohydrates. 8-aminopyrene-1,3,6-trisulfonic acid (APTS) was used for the labeling of the carbohydrate standards and honey sugars for fluorescence detection. The optimized method was applied in the quantitative analysis of fructose and glucose by direct injection of honey samples. Apart from the labeling reaction, no other sample preparation was performed. The mean values of the fructose/glucose ratio for phacelia honey, acacia honey and honeydew honey were 0.86, 1.61 and 1.42, respectively. The proposed method provides high separation efficiency and sensitive detection within a short analysis time. Apart from the labeling reaction, it enables the injection of honeys without sample pretreatment. This is the first time that fluorescence detection has been applied for the CE analysis of sugars in honeys.

Setophoma terrestris is an important phytopathogenic fungus listed by China as a harmful fungus subject to phytosanitary import control. This pathogen is a threat to a wide range of plants, particularly as the causal agent of onion pink root rot, one of the most severe diseases of onions. In order to provide rapid identification and early warning of S. terrestris and prevent its spread, we have developed a rapid, accurate, and visually intuitive diagnostic assay for this pathogen, by utilizing recombinase polymerase amplification (RPA), coupled with CRISPR/Cas12a cleavage and fluorescence-based detection systems or paper-based lateral flow strips. The developed RPA-CRISPR/Cas12a assay exhibited remarkable specificity for the detection of S. terrestris. Moreover, this protocol can detect the pathogen at a sensitivity level of 0.01 pg/μL, which significantly outperforms the 1 pg/μL sensitivity achieved by the existing qPCR-based detection method. The entire diagnostic procedure, including DNA extraction, the RPA reaction, the Cas12a cleavage, and the result interpretation, can be accomplished in 40 min. Furthermore, the successful application of the assay in infected plant samples highlighted its potential for rapid and accurate pathogen detection in agricultural settings. In summary, this RPA-CRISPR/Cas12a diagnostic method offers a potentially valuable technological solution for quarantine and disease management.

Rose Bengal (RB) is a fluorescent dye with several potential biomedical applications, particularly in dermatology. Due to RB’s poor physicochemical properties, several advanced delivery systems have been developed as a potential tool to promote its permeation across the skin. Nevertheless, no validated quantitative method to analyse RB within the skin is described in the literature. Considering RB exhibits a conjugated ring system, the current investigation proposes fluorescence-based techniques beneficial for qualitatively and quantitatively determining RB delivered to the skin. Notably, the development and validation of a fluorescence-coupled HPLC method to quantify RB within the skin matrix are herein described for the first time. The method was validated based on the ICH, FDA and EMA guidelines, and the validated parameters included specificity, linearity, LOD, LLOQ, accuracy and precision, and carry-over and dilution integrity. Finally, the method was applied to evaluate RB’s ex vivo permeation and deposition profiles when loaded into dermatological formulations. Concerning qualitative determination, multiphoton microscopy was used to track the RB distribution within the skin strata, and fluorescence emission spectra were investigated to evaluate RB’s behaviour when interacting with different environments. The analytical method proved specific, precise, accurate and sensitive to analyse RB in the skin. In addition, qualitative side-analytical techniques were revealed to play an essential role in evaluating the performance of RB’s dermatological formulation.

Machine learning and image processing have been combined to identify and detect defects in mature citrus fruit at night, which has great research and development significance. First, a multi-light vision system was used to collect citrus UV images, and from these, 1500 samples were obtained, 80% of which were training and 20% were experimental sets. For a support vector machine (SVM) model with “2*Cb-Cr”, “4*a-b-l”, and “H” as the training features, the accuracy of the final training model in the experimental set is 99.67%. Then, the SVM model was used to identify mature citrus regions, detect defects, and output the defective citrus regions label. The average running time of the detection algorithm was 0.84097 s, the accuracy of citrus region detection was 95.32%, the accuracy of citrus defect detection was 96.32%, the precision was 95.24%, and the recall rate was 87.91%. The results show that the algorithm had suitable accuracy and real-time performance in recognition and defect detection in citrus in a natural environment at night.

In this work, we present a multifunctional Lab-on-Chip (LoC) platform based on hydrogenated amorphous silicon sensors suitable for a wide range of application in the fields of biochemical and food quality control analysis. The proposed system includes a LoC fabricated on a 5 cm × 5 cm glass substrate and a set of electronic boards for controlling the LoC functionalities. The presented Lab-on-Chip comprises light and temperature sensors, a thin film resistor acting as a heating source, and an optional thin film interferential filter suitable for fluorescence analysis. The developed electronics allows to control the thin film heater, a light source for fluorescence and absorption measurements, and the photosensors to acquire luminescent signals. All these modules are enclosed in a black metal box ensuring the portability of the whole platform. System performances have been evaluated in terms of sensor optical performances and thermal control achievements. For optical sensors, we have found a minimum number of detectable photons of 8 × 104 s−1·cm−2 at room temperature, 1.6 × 106 s−1·cm−2 in presence of fluorescence excitation source, and 2.4 × 106 s−1·cm−2 at 90 °C. From a thermal management point of view, we have obtained heating and cooling rates both equal to 2.2 °C/s, and a temperature sensor sensitivity of about 3 mV/°C even in presence of light. The achieved performances demonstrate the possibility to simultaneously use all integrated sensors and actuators, making promising the presented platform for a wide range of application fields.

The polymerase chain reaction is an important technique in biological research. However, it is time consuming and has a number of disadvantages. Therefore, real-time PCR technology that can be used in real-time monitoring has emerged, and many studies are being conducted regarding its use. Real-time PCR requires many optical components and imaging devices such as expensive, high-performance cameras. Therefore, its cost and assembly process are limitations to its use. Currently, due to the development of smart camera devices, small, inexpensive cameras and various lenses are being developed. In this paper, we present a Compact Camera Fluorescence Detector for use in parallel-light lens-based real-time PCR devices. The proposed system has a simple optical structure, the system cost can be reduced, and the size can be miniaturized. This system only incorporates Fresnel lenses without additional optics in order for the same field of view to be achieved for 25 tubes. In the center of the Fresnel lens, one LED and a complementary metal-oxide semiconductor camera were placed in directions that were as similar as possible. In addition, to achieve the accurate analysis of the results, image processing was used to correct them. As a result of an experiment using a reference fluorescent substance and double-distilled water, it was confirmed that stable fluorescence detection was possible.

This study proposed a filter-free wavelength sensor with a double-well structure for detecting fluorescence without an optical filter. The impurity concentration was optimized and simulated to form a double-well-structured sensor, of which the result was consistent with the fabricated sensor. Furthermore, we proposed a novel wavelength detection method using the current ratio based on the silicon absorption coefficient. The results showed that the proposed method successfully detected single wavelengths in the 460–800 nm range. Additionally, we confirmed that quantification was possible using the current ratio of the sensor for a relatively wide band wavelength, such as fluorescence. Finally, the fluorescence that was emitted from the reagents ALEXA488, 594, and 680 was successfully identified and quantified. The proposed sensor can detect wavelengths without optical filters, which can be used in various applications in the biofield, such as POCT as a miniaturized wavelength detection sensor.

The combination of a metallated porphyrin, Pt(II)-5,10,15,20-tetrakis-(4-allyloxyphenyl)-porphyrin (Pt-allyloxyPP), and a water-soluble porphyrin, 5,10,15,20-tetrakis(4-sulfonatophenyl)-porphyrin (TSPP), leads to the formation of a porphyrin hetero-trimer. The hetero-trimer, consisting of two TSPP molecules linked via oxygen atoms axially to the platinum atom in the Pt-allyloxyPP molecule, was characterized by UV–Vis, FT-IR, fluorescence, and 1H-NMR spectroscopy, and the proposed structure was confirmed. The new porphyrin hetero-trimer offers both the advantage of enhanced fluorescence and the presence of multiple sites for the detection of toluidine blue, due to its high affinity for acidic binding sites. This work brings attention to the purposely designed fluorescent sensor for toluidine blue, in the biologically relevant concentration domain of 1.9 × 10−6–6.39 × 10−5 M, with a very good accuracy.

Sudan dyes are strictly prohibited from being added to edible products as carcinogens and tetracycline hydrochloride (TC) remaining in animal-derived food may cause harm to the human body. Therefore, it is necessary to establish a high-sensitivity, simple and convenient method for the detection of Sudan dyes and TC in foods for safety purposes. In this work, multifunctional blue fluorescent carbon dots (B-CDs) were prepared by a one-step hydrothermal synthesis using glucose as the carbon source. The results show that the fluorescence intensity of B-CDs was significantly affected by the acidity of the solution and can be quenched by Sudan I, IV and TC through selective studies. Interestingly, the fluorescence quenching intensities of B-CDs have a good linear relationship with the concentration of Sudan I and IV at pH = 3–7. The wide range of pH is beneficial to broaden the application of B-CDs in a practical samples analysis. The method has been successfully applied to real food samples of tomato paste, palm oil and honey, and the detection limits are 26.3 nM, 54.2 nM and 31.1 nM for Sudan I, Sudan IV and TC, respectively. This method integrates Sudan dyes and TC into the same multifunctional B-CDs, which shows that the sensor has a great potential in food safety detection.

An enzyme-linked fluorescent immunoassay (ELFIA) method has been developed for the quantitative analytical determination of the herbicide active ingredient glyphosate in environmental matrices (surface water, soil, and plant tissues). Glyphosate, as a ubiquitous agricultural pollutant, is a xenobiotic substance with exposure in aquatic and terrestrial ecosystems due its extremely high worldwide application rate. The immunoassay developed in Project Aquafluosense is part of a fluorescence-based instrumentation setup for the in situ determination of several characteristic water quality parameters. The 96-well microplate-based competitive immunoassay method applies fluorescence signal detection in the concentration range of 0–100 ng/mL glyphosate. Application of the fluorescent signal provides a limit of detection of 0.09 ng/mL, which is 2.5-fold lower than that obtained with a visual absorbance signal. Beside the improved limit of detection, determination by fluorescence provided a wider and steeper dynamic range for glyphosate detection. No matrix effect appeared for the undiluted surface water samples, while plant tissues and soil samples required dilution rates of 1:10 and 1:100, respectively. No cross-reaction was determined with the main metabolite of glyphosate, N-aminomethylphosphonic acid, and related compounds.

Herein, we present a comprehensive investigation of rationally designed zinc selenide (ZnSe) nanostructures to achieve highly negatively charged ZnSe nanostructures. A Microwave-assisted hydrothermal synthesis method was used to synthesize three types of ZnSe nanostructures, i.e., nanorods, µ-spheres and nanoclusters, as characterized by a zeta potential analyzer, X-ray diffraction (XRD), scanning electron microscopy (SEM), Raman spectroscopy and BET, which were labeled as type A, B and C. Three different solvents were used for the synthesis of type A, B and C ZnSe nanostructures, keeping other synthesis conditions such as temperature, pressure and precursors ratio constant. Based on two heating time intervals, 6 and 9 h, types A, B and C were further divided into types A6, A9, B6, B9, C6 and C9. ZnSe nanostructures were further evaluated based on their fluorescent quenching efficiency. The maximum fluorescence quenching effect was exhibited by the ZnSe-B6 type, which can be attributed to its highly negative surface charge that favored its strong interaction with cationic dye Rhodamine B (Rh-B). Further, the optimized ZnSe-B6 was used to fabricate an aptasensor for the detection of a food-based toxin, ochratoxin-A (OTA). The developed aptasensor exhibited a limit of detection of 0.07 ng/L with a wide linear range of 0.1 to 200 ng/L.

In this work, we examine how strain exerted on individual ZnO nanorods (NRs) can influence the fluorescence signals that are emitted from fluorophore molecules and subsequently coupled into and guided along the NR. We elucidate the relationships between the incremental levels of compressive and tensile strain on the NRs and measured fluorescence intensity of a model fluorophore, rhodamine 6G (R6G), as a function of the position on the NRs. We reveal that compressive strain on the NRs leads to a decrease in the guided fluorescence signal, while tensile strain leads to an increase in the fluorescence intensity. Compared to an unstrained state, approximately 35% decrease (increase) in R6G fluorescence intensity was observed from ZnO NRs when they were under compressive strain of −14% (tensile strain of +10%). Further, our systematic acquisition of the incremental addition of uniaxial strain result in a linear relationship of the coupled fluorescence signal and the amount of applied strain. The degree of fluorescence intensification on nanorod ends (DoF), which is a quantitative indicator for the amount of R6G signals coupled into and waveguided to the NR ends compared to those on the main body, also exhibits a linear relationship with strain. These outcomes, in turn, demonstrate that strain alters the waveguiding capabilities of ZnO NRs in a predictable manner, which can be exploited to modulate and optimize fluorescence and other light signals emitted by a nearby source. Considering the wide utility of ZnO NRs in photonics, optoelectronics, and sensors, insights from our study may be highly valuable to effectively controlling and enhancing optical signals from chemical and biological analytes through strain.

Acetaldehyde is regarded as an important flavor compound in alcoholic beverages. With the advantages of rapidity, low cost and high sensitivity, fluorescent probe could be used as a new tool for the detection of acetaldehyde. Here, an effective fluorescence sensing method based on fluorescent probe N1 (FPN1) was established in this study. The function of FPN1 relies on the nucleophile substitution reaction and photoinduced electron transfer (PET), resulting in a fluorescence increase. Remarkably, the pretreatment background removal method (BRM) was successfully applied for removal of the interference of pyruvate and acetal. The linearity range (LR), limit of detection (LOD) and recovery of the fluorescence sensing method with BRM were 0.0053–200 mg/L, 0.0016 mg/L and 94.02–108.12%, respectively, which showed a broader detection range and better performance on sensitivity compared with the traditional quantitation using gas chromatography (GC). Furthermore, successful application of the method in real samples indicated the advantages of low-cost and rapidity for small-scale detection while assuring the accuracy, which provides a new strategy for the detection of acetaldehyde concentration in alcoholic beverages.

The COVID-19 pandemic poses a threat to global health. Due to its high sensitivity, specificity, and stability, real-time fluorescence quantitative (real-time PCR) detection has become the most extensively used approach for diagnosing SARS-CoV-2 pneumonia. According to a report from the World Health Organization, emerging and underdeveloped nations lack nucleic acid detection kits and polymerase chain reaction (PCR) instruments for molecular biological detection. In addition, sending samples to a laboratory for testing may result in considerable delays between sampling and diagnosis, which is not favorable to the timely prevention and control of new crown outbreaks. Concurrently, there is an urgent demand for accurate PCR devices that do not require a laboratory setting, are more portable, and are capable of completing testing on-site. Hence, we report on HDLRT-qPCR, a new, low-cost, multiplexed real-time fluorescence detection apparatus that we have developed for on-site testing investigations of diverse diseases in developing nations. This apparatus can complete on-site testing rapidly and sensitively. The entire cost of this instrument does not exceed USD 760. In order to demonstrate the applicability of our PCR instrument, we conducted testing that revealed that we achieved gradient amplification and melting curves comparable to those of commercially available equipment. Good consistency characterized the testing outcomes. The successful detection of target genes demonstrates the reliability of our inexpensive PCR diagnostic technique. With this apparatus, there is no need to transport samples to a central laboratory; instead, we conduct testing at the sampling site. This saves time on transportation, substantially accelerates overall testing speed, and provides results within 40 min.

As an important detection tool in biochemistry, fluorescence detection has wide applications. Quantitative detection can be achieved by detecting fluorescence signals excited by excitation light at a specific wavelength range. Therefore, the key to fluorescence detection is the stable control of the excitation light and the accurate acquisition of weak photoelectric signals. Moreover, to improve portability and instantaneity, devices are developing in miniaturization and integration. As the core of such devices, fluorescence detectors should also have these features. Under this circumstance, we designed a highly integrated and diminutive fluorescence detector and focused on its excitation light driving and photoelectric signal processing. A current–light dual negative feedback light-emitting diode (LED) driving circuit was proposed to obtain constant current and luminance. In addition, a silicon photodiode (PD) was used to receive and convert the fluorescence signal to an electric signal. Then, amplifying, filtering, and analog-to-digital (A/D) converting were applied to make the detection of weak fluorescence signals possible. The test results showed that the designed circuit has wonderful performance, and the detector shows good linearity (R2 = 0.9967) and sensitivity (LOD = 0.077 nM) in the detection of fluorescein sodium solution. Finally, a real-time fluorescence polymerase chain reaction (real-time PCR) of Legionella pneumophila was carried out on a homemade platform equipped with this detector, indicating that the detector met the requirements of real-time PCR detection.

Two water-soluble single-benzene-based chromophores, 2,5-di(azetidine-1-yl)-tereph- thalic acid (DAPA) and its disodium carboxylate (DAP-Na), were conveniently obtained. Both chromophores preserved moderate quantum yields in a wide range of polar and protonic solvents. Spectroscopic studies demonstrated that DAPA exhibited red luminescence as well as large Stokes shift (>200 nm) in aqueous solutions. Femtosecond transient absorption spectra illustrated quadrupolar DAPA usually involved the formation of an intramolecular charge transfer state. Its Frank–Condon state could be rapidly relaxed to a slight symmetry-breaking state upon light excitation following the solvent relaxation, then the slight charge separation may occur and the charge localization became partially asymmetrical in polar environments. Density functional theory (DFT) calculation results were supported well with the experimental measurements. Unique pH-dependent fluorescent properties endows the two chromophores with rapid, highly selective, and sensitive responses to the amino acids in aqueous media. In detail, DAPA served as a fluorescence turn-on probe with a detection limit (DL) of 0.50 μM for Arg and with that of 0.41 μM for Lys. In contrast, DAP-Na featured bright green luminescence and showed fluorescence turn-off responses to Asp and Glu with the DLs of 0.12 μM and 0.16 μM, respectively. Meanwhile, these two simple-structure probes exhibited strong anti-interference ability towards other natural amino acids and realized visual identification of specific analytes. The present work helps to understand the photophysic–structure relationship of these kinds of compounds and render their fluorescent detection applications.

The fabrication of efficient fluorescent probes that possess an excellent sensitivity and selectivity for uric acid is highly desirable and challenging. In this study, composites of silver nanoparticles (AgNPs) wrapped with nitrogen-doped carbon quantum dots (N-CQDs) were synthesised utilising N-CQDs as the reducing and stabilising agents in a single reaction with AgNO3. The morphology and structure, absorption properties, functional groups, and fluorescence properties were characterised by transmission electron microscopy, X-ray photoelectron spectroscopy, Fourier transform infrared spectroscopy, ultraviolet spectroscopy, fluorescence spectroscopy, and X-ray diffraction spectroscopy. In addition, we developed a novel method based on AgNPs/N-CQDs for the detection of uric acid using the enzymatic reaction of uric acid oxidase. The fluorescence enhancement of the AgNPs/N-CQDs composite was linear (R2 = 0.9971) in the range of 2.0–60 μmol/L, and gave a detection limit of 0.53 μmol/L. Trace uric acid was successfully determined in real serum samples from the serum of 10 healthy candidates and 10 gout patients, and the results were consistent with those recorded by Qianxinan Prefecture People’s Hospital. These results indicate that the developed AgNP/N-CQD system can provide a universal platform for detecting the multispecies ratio fluorescence of H2O2 generation in other biological systems.

Vibrio parahaemolyticus is one of the major pathogenic Vibrio species that contaminate seafood. Rapid and accurate detection is crucial for avoiding foodborne diseases caused by pathogens and is important for food safety management and mariculture. In this study, we established a system that combines chemically enhanced clustered regularly interspaced short palindromic repeats (CRISPR) and recombinase-aided amplification (RAA) (CE–RAA–CRISPR) for detecting V. parahaemolyticus in seafood. The method combines RAA with CRISPR-associated protein 12a (Cas12a) for rapid detection in a one-pot reaction, effectively reducing the risk of aerosol contamination during DNA amplifier transfer. We optimized the primers for V. parahaemolyticus, determined the optimal crRNA/Cas12a ratio, and demonstrated that chemical additives (bovine serum albumin and L-proline) could enhance the detection capacity of Cas12a. The limit of detection (at optimal conditions) was as low as 6.7 × 101 CFU/mL in pure cultures and 7.3 × 101 CFU/g in shrimp. Moreover, this method exhibited no cross-reactivity with other microbial pathogens. The CE–RAA–CRISPR assay was compared with the quantitative polymerase chain reaction assay using actual food samples, and it showed 100% diagnostic agreement.

Traditional qPCR instrument is combined with CMOS and a personal computer, and a photoelectric feedback automatic fluorescence detection system is designed to realize quantitative real-time PCR. The key to reaction efficiency lies in how to ensure that the temperature of the detection reagent completely matches the set temperature. However, for most traditional real-time fluorescent PCR systems, the temperature cycling is controlled by detecting the temperature of the heating well plate. It cannot directly measure the temperature in the reaction reagent PCR tube, which will cause the deviation in the actual temperature of the reagent to be as expected. Therefore, in this paper, we raise a method of directly detecting the temperature in the reaction tube of the reagent during the temperature cycling is adopted. According to the deviation from the expected value, the set temperature of the PCR instrument is adjusted to make the actual temperature of the reagent closer to the expected value. Through this method, we also realized the temperature calibration and optimization of the TEC circulation system we built. Experiments show that this low-cost, portable real-time quantitative PCR system can detect and analyze pathogens in situ.

Chlorella vulgaris is of great importance in numerous exploratory or industrial applications (e.g., medicals, food, and feed additives). Rapid quantification of algal biomass is crucial in photobioreactors for the optimization of nutrient management and the estimation of production. The main goal of this study is to provide a simple, rapid, and not-resource-intensive estimation method for determining the algal density of C. vulgaris according to the measured parameters using UV-Vis spectrophotometry. Comparative assessment measurements were conducted with seven different methods (e.g., filtration, evaporation, chlorophyll a extraction, and detection of optical density and fluorescence) to determine algal biomass. By analyzing the entire spectra of diluted algae samples, optimal wavelengths were determined through a stepwise series of linear regression analyses by a novel correlation scanning method, facilitating accurate parameter estimation. Nonlinear formulas for spectrometry-based estimation processes were derived for each parameter. As a result, a general formula for biomass concentration estimation was developed, with recommendations for suitable measuring devices based on algae concentration levels. New values for magnesium content and the average single-cell weight of C. vulgaris were established, in addition to the development of a rapid, semiautomated cell counting method, improving efficiency and accuracy in algae quantification for cultivation and biotechnology applications.

African swine fever (ASF) is a swine disease with a very high fatality rate caused by a complex double-stranded DNA virus. The fluorescence PCR detection method is widely used for virus nucleic acid detection. Surface plasmon resonance (SPR) is a label-free and real-time detection method, unlike the fluorescence PCR detection method. In this research, we detected the loop-mediated isothermal amplification (LAMP) products of the African swine fever virus by using the SPR and fluorescence methods separately and simultaneously. By comparing the positive and negative control results, we found that the SPR response unit is completely different before and after the LAMP process. In addition, the fluorescence results on a chip showed that with an increase in the concentration of the sample, the cycle threshold (CT) value decreased, which is consistent with commercial instruments. Both the decline rate of the SPR response unit and the CT value of the fluorescence realized were used to distinguish the positive control from the negative control and water, which indicates that the SPR method can be combined with fluorescence to detect LAMP products. This research provides a label-free and simple method for detecting LAMP products.

Hollow carbon nitride nanosphere (HCNS) was synthesized via the hard template method to improve the fluorescence characteristics, drug delivery ability, and photocatalytic activity. Blue fluorescent HCNS was utilized as a quenching agent and an internal reference to combine with Cy5-labelled aptamer (Cy5-Apt), resulting in an off-on fluorescence aptasensing method for the detection of Salmonella typhimurium (S. typhimurium). Under optimum conditions, this fluorescence assay presented a linear range from 30 to 3 × 104 CFU mL−1 with a detection limit of 13 CFU mL−1. In addition, HCNS was also used as a drug carrier to load chloramphenicol (Cap) molecules. The Cap-loading amount of HCNS could reach 550 μg mg−1 within 24 h, whereas the corresponding Cap-release amount is 302.5 μg mg−1 under acidic and irradiation conditions. The integration of photocatalyst with antibiotic could endow HCNS-Cap with better disinfection performance. The bactericidal efficiency of HCNS-Cap (95.0%) against S. typhimurium within 12 h was better than those of HCNS (85.1%) and Cap (72.9%). In addition, selective disinfection of S. typhimurium was further realized by decorating aptamer. Within 4 h, almost all S. Typhimurium were inactivated by HCNS-Cap-Apt, whereas only 13.3% and 48.2% of Staphylococcus aureus and Escherichia coli cells were killed, respectively. Therefore, HCNS is a promising bio-platform for aptamer-based fluorescence detection and selective disinfection of S. typhimurium.

We introduce a digital microfluidics (DMF) platform specifically designed to perform a loop-mediated isothermal amplification (LAMP) of DNA and applied it to a real-time amplification to monitor a cancer biomarker, c-Myc (associated to 40% of all human tumors), using fluorescence microscopy. We demonstrate the full manipulation of the sample and reagents on the DMF platform, resulting in the successful amplification of 90 pg of the target DNA (0.5 ng/µL) in less than one hour. Furthermore, we test the efficiency of an innovative mixing strategy in DMF by employing two mixing methodologies onto the DMF droplets—low frequency AC (alternating current) actuation as well as back-and-forth droplet motion—which allows for improved fluorescence readouts. Fluorophore bleaching effects are minimized through on-chip sample partitioning by DMF processes and sequential droplet irradiation. Finally, LAMP reactions require only 2 µL volume droplets, which represents a 10-fold volume reduction in comparison to benchtop LAMP.

Nitrogen-doped carbon dots exhibiting excitation-dependent full-color emissions (F-NCDs) were prepared via the one-step hydrothermal method with citric acid and phenylenediamine. Specifically, the emission wavelength of the F-NCDs tuned from 452 nm to 602 nm due to the introduction of new energy levels by C=O and C=N functional groups. We exploited its stability in illumination, ionic strength, and pH, as well as its specificity, sensitivity, especially in ascorbic acid (AA) detection. F-NCDs could measure the AA concentration in the linear ranges of 0~0.1 and 0.1~1 mmol/L with the detection limit (LOD, S/N = 3) as low as 2.6 nmol/L. Additionally, we successfully detected AA in bovine serum with our F-NCDs and obtained the result within 1 min. Because of full-color emission features, we believe our F-NCDs have a great potential in fluorescent sensor detection.

Biological water contamination detection-based assays are essential to test water quality; however, these assays are prone to false-positive results and inaccuracies, are time-consuming, and use complicated procedures to test large water samples. Herein, we show a simple detection and counting method for E. coli in the water samples involving a combination of DNAzyme sensor, microfluidics, and computer vision strategies. We first isolated E. coli into individual droplets containing a DNAzyme mixture using droplet microfluidics. Upon bacterial cell lysis by heating, the DNAzyme mixture reacted with a particular substrate present in the crude intracellular material (CIM) of E. coli. This event triggers the dissociation of the fluorophore-quencher pair present in the DNAzyme mixture leading to a fluorescence signal, indicating the presence of E. coli in the droplets. We developed an algorithm using computer vision to analyze the fluorescent droplets containing E. coli in the presence of non-fluorescent droplets. The algorithm can detect and count fluorescent droplets representing the number of E. coli present in the sample. Finally, we show that the developed method is highly specific to detect and count E. coli in the presence of other bacteria present in the water sample.

Eating vegetables with pesticide residues over a long period of time causes serious adverse effects on the human body, such as acute poisoning, chronic poisoning, and endocrine system interference. To achieve the goal of a healthy society, it is an urgent issue to find a simple and effective method to detect organic pesticides. In this work, two fluorescent porous organic polymers, LNU-45 and LNU-47 (abbreviation for Liaoning University), were prepared using π-conjugated dibromopyrene monomer and boronic acid compounds as building units through a Suzuki coupling reaction. Due to the large π-electron delocalization effect, the resulting polymers revealed enhanced fluorescence performance. Significantly, in sharp contrast with the planar π-conjugated polymer framework (LNU-47), the distorted conjugated structure (LNU-45) shows a higher specific surface area and provides a broad interface for analyte interaction, which is helpful to achieve rapid response and detection sensitivity. LNU-45 exhibits strong fluorescence emission at 469 nm after excitation at 365 nm in THF solution, providing strong evidence for its suitability as a luminescent chemosensor for organic pesticides. The fluorescence quenching coefficients of LNU-45 for trifluralin and dicloran were 5710 and 12,000 (LNU-47 sample by ca. 1.98 and 3.38 times), respectively. Therefore, LNU-45 serves as an effective “real-time” sensor for the detection of trifluralin and dicloran with high sensitivity and selectivity.

Biogenic polyamines, especially spermine and spermidine, are associated with cell growth and development. These amines can be found at high concentrations in the tumor cells, tissues, and urine of cancer patients. In contrast, spermidine levels drop with age, and a possible connection between low endogenous spermidine concentrations and age-related deterioration has been suggested. Thus, the quantification of these amines in body fluids like urine could be used in the diagnosis of different pathological situations. Here a new fluorescent molecular probe based on a tetraphenylethylene derivative is reported. This probe is able to selectively detect these amines through the enhancement of the fluorescence emission of the resulting complex. This fluorescence enhancement may be related to restricted intramolecular rotations of TPE phenyl rings induced by the analyte. Theoretical studies were carried out to shed light on the observed selectivity. Finally, the detection of these amines in urine was performed with limits of detection of 0.70 µM and 1.17 µM for spermine and spermidine, respectively.

The fast spread of SARS-CoV-2 has led to a global pandemic, calling for fast and accurate assays to allow infection diagnosis and prevention of transmission. We aimed to develop a molecular beacon (MB)-based detection assay for SARS-CoV-2, designed to detect the ORF1ab and S genes, proposing a two-stage COVID-19 testing strategy. The novelty of this work lies in the design and optimization of two MBs for detection of SARS-CoV-2, namely, concentration, fluorescence plateaus of hybridization, reaction temperature and real-time results. We also identify putative G-quadruplex (G4) regions in the genome of SARS-CoV-2. A total of 458 nasopharyngeal and throat swab samples (426 positive and 32 negative) were tested with the MB assay and the fluorescence levels compared with the cycle threshold (Ct) values obtained from a commercial RT-PCR test in terms of test duration, sensitivity, and specificity. Our results show that the samples with higher fluorescence levels correspond to those with low Ct values, suggesting a correlation between viral load and increased MB fluorescence. The proposed assay represents a fast (total duration of 2 h 20 min including amplification and fluorescence reading stages) and simple way of detecting SARS-CoV-2 in clinical samples from the upper respiratory tract.

The polymerase chain reaction is an important technique in biological research because it tests for diseases with a small amount of DNA. However, this process is time consuming and can lead to sample contamination. Recently, real-time PCR techniques have emerged which make it possible to monitor the amplification process for each cycle in real time. Existing camera-based systems that measure fluorescence after DNA amplification simultaneously process fluorescence excitation and emission for dozens of tubes. Therefore, there is a limit to the size, cost, and assembly of the optical element. In recent years, imaging devices for high-performance, open platforms have benefitted from significant innovations. In this paper, we propose a fluorescence detector for real-time PCR devices using an open platform camera. This system can reduce the cost, and can be miniaturized. To simplify the optical system, four low-cost, compact cameras were used. In addition, the field of view of the entire tube was minimized by dividing it into quadrants. An effective image processing method was used to compensate for the reduction in the signal-to-noise ratio. Using a reference fluorescence material, it was confirmed that the proposed system enables stable fluorescence detection according to the amount of DNA.

Laser-induced fluorescence (LIF) has been a valuable analytical technique since the 1970s that has only been made more useful through advances in other scientific fields such as biochemistry. Moreover, advances in laser and detector technology have seen a decrease in LIF detector costs and an increase in their ease of use. These changes have allowed for LIF technology to be widely adopted for various sensor designs in combination with advanced instruments. With advances in biochemistry necessitating the detection of complex metabolites, labelling with fluorescent chemical reagents may be necessary to improve detection sensitivity. Furthermore, advances made in fluorescent labeling technologies have allowed for the use of LIF in the detection of nanoparticles as well as for imaging techniques using nanoparticles as signal amplifiers. This technology has become invaluable in the detection of environmental pollutants, monitoring of biological metabolites, biological imaging, and cancer diagnosis, making it one of the most valuable analytical science techniques currently available.