Search Results (58)

Desulfovibrio vulgaris is an anaerobic microorganism belonging to the group of sulphate-reducing bacteria (SRB). SRB form biofilms on metal surfaces in water supply networks, producing a microbiologically influenced corrosion (MIC). This process produces the deterioration of metal surfaces, leading to high economic costs and different environmental safety and health problems related to its chemical treatment. For that reason, rapid and accurate detection methods of SRB are needed. In this work, a new detection system for Desulfovibrio has been developed using gated nanoporous materials. The probe is based on hybrid nanoporous alumina films encapsulating a fluorescent molecule (rhodamine B), whose release is controlled by an oligonucleotide gate. Upon exposure to Desulfovibrio’s genomic material, a movement of the oligonucleotide gatekeeper happens, resulting in the selective delivery of the entrapped rhodamine B. The developed material shows high selectivity and sensitivity for detecting Desulfovibrio DNA in aqueous buffer and biological media. The implementation of this technology for the detection of Desulfovibrio as a tool for monitoring water supply networks is innovative and allows real-time in situ monitoring, making it possible to detect the growth of Desulfovibrio inside of pipes at an early stage and perform timely interventions to reverse it. Full article

Voltage-gated sodium (Nav) channels play a crucial role in initiating and propagating action potentials throughout the heart, muscles and nervous systems, making them targets for a number of drugs and toxins. While patch-clamp electrophysiology is considered the gold standard for measuring ion channel activity, its labor-intensive and time-consuming nature highlights the need for fast screening strategies to facilitate a preliminary selection of potential drugs or hazards. In this study, a high-throughput and cost-effective biosensing method was developed to rapidly identify specific agonists and inhibitors targeting the human Nav1.1 (hNav1.1) channel. It combines a red fluorescent dye sensitive to transmembrane potentials with CHO cells stably expressing the hNav1.1 α-subunit (hNav1.1-CHO). In the initial screening mode, the tested compounds were mixed with pre-equilibrated hNav1.1-CHO cells and dye to detect potential agonist effects via fluorescence enhancement. In cases where no fluorescence enhancement was observed, the addition of a known agonist veratridine allowed the indication of inhibitor candidates by fluorescence reduction, relative to the veratridine control without test compounds. Potential agonists or inhibitors identified in the initial screening were further evaluated by measuring concentration–response curves to determine EC₅₀/IC₅₀ values, providing semi-quantitative estimates of their binding strength to hNav1.1. This robust, high-throughput biosensing assay was validated through comparisons with the patch-clamp results and tested with 12 marine toxins, yielding consistent results. It holds promise as a low-cost, rapid, and long-term stable approach for drug discovery and non-target screening of neurotoxins. Full article

Telomeres play a key role in maintaining chromosome stability and cellular aging. They consist of repetitive DNA sequences that protect chromosome ends and regulate cell division. Telomerase is a reverse transcriptase enzyme counteracts the natural shortening of telomeres during cell division by extending them. Its activity is pivotal in stem cells and cancer cells but absent in most normal somatic cells. Recent advances in biosensor technologies have facilitated the in situ detection of telomerase activity, which is essential for understanding its role in aging and cancer. Techniques such as fluorescence, electrochemistry, and DNA nanotechnology are now being employed to monitor telomerase activity in living cells, providing real-time insights into cellular processes. DNA-based biosensors, especially those incorporating molecular beacons, DNA walkers, and logic gates, have shown promise for enhancing sensitivity and specificity in telomerase imaging. These approaches also facilitate the simultaneous analysis of related cellular pathways, offering potential applications in early cancer detection and precision therapies. This review explores recent developments in intracellular telomerase imaging, highlighting innovative approaches such as DNA-functionalized nanoparticles and multi-channel logic systems, which offer non-invasive, real-time detection of telomerase activity in complex cellular environments. Full article

Fluorescence imaging has been widely used in fields like (pre)clinical imaging and other domains. With advancements in imaging technology and new fluorescent labels, fluorescence lifetime imaging is gradually gaining recognition. Our research department is developing the tauCAM^TM, based on the Current-Assisted Photonic Sampler, to achieve real-time fluorescence lifetime imaging in the NIR (700–900 nm) region. Incorporating fluorescence lifetime into endoscopy could further improve the differentiation of malignant and benign cells based on their distinct lifetimes. In this work, the capabilities of an endoscopic lifetime imaging system are demonstrated using a rigid endoscope involving various phantoms and an IRF-free deep learning-based method with only 6-time points. The results show that this application’s fluorescence lifetime image has better lifetime uniformity and precision with 6-time points than the conventional methods. Full article

We report super broad non-Hermitian line shape from out-of-phase and in-phase photon-phonon dressing (quantization) in Eu³⁺: NaYF₄ and Eu³⁺: BiPO₄ nanocrystals. The line shape is controlled by changing time gate position, time gate width, power, temperature, sample, photomultiplier tubes, and laser. We observed that the fluorescence (FL) line-shape contrasts are 69.23% for Eu³⁺: BiPO₄ and 43.75% for Eu³⁺: NaYF₄, owing to the stronger out-of-phase photon-phonon dressing (destructive quantization). Moreover, we observed that the spontaneous four-wave mixing (SFWM) line shape was approximately three times wider at 300 K than at 77 K for the [(12:1)-phase] Eu³⁺: NaYF₄ due to more high-frequency in-phase phonon dressing (strong constructive quantization). Furthermore, we showed that the noise line-shape width remains unchanged for Eu³⁺: BiPO₄ (16 nm) and Eu³⁺: NaYF₄ (12 nm) due to out-of-phase and in-phase photon-phonon dressing balance. Such results have potential applications in multi-channel band stop filter. Full article

Sepsis, a life-threatening condition resulting from a failing host response to infection, causes millions of deaths annually, necessitating rapid and simple prognostic assessments. A variety of genomic and proteomic biomarkers have been developed for sepsis. For example, it has been shown that the level of plasma cell-free DNA (cfDNA) and circulating histones increases considerably during sepsis, and they are linked with sepsis severity and mortality. Developing a diagnostic tool that is capable of assessing such diverse biomarkers is challenging as the detection methodology is quite different for each. Here, a fully integrated microfluidic device capable of detecting a genomic biomarker (cfDNA) and a proteomic biomarker (total circulating histones) using a common detection platform has been demonstrated. The microfluidic device utilizes dehydrated agarose gates loaded with pH-specific agarose to electrophoretically trap cfDNA and histones at their respective isoelectric points. It also incorporates fluorescent dyes within the device, eliminating the need for off-chip sample preparation and allowing the direct testing of plasma samples without the need for labeling DNA and histones with fluorescent dyes beforehand. Xurography, which is a low-cost and rapid method for fabrication of microfluidics, is used in all the fabrication steps. Experimental results demonstrate the effective accumulation and separation of cfDNA and histones in the agarose gates in a total processing time of 20 min, employing 10 and 30 Volts for cfDNA and histone accumulation and detection, respectively. The device can potentially be used to distinguish between the survivors and non-survivors of sepsis. The integration of the detection of both biomarkers into a single device and dye immobilization enhances its clinical utility for rapid point-of-care assessment of sepsis prognosis. Full article

Rapid detection of fish freshness is of vital importance to ensuring the safety of aquatic product consumption. Currently, the widely used optical detecting methods of fish freshness are faced with multiple challenges, including low detecting efficiency, high cost, large size and low integration of detecting equipment. This research aims to address these issues by developing a low-cost portable fluorescence imaging device for rapid fish freshness detection. The developed device employs ultraviolet-light-emitting diode (UV-LED) lamp beads (365 nm, 10 W) as excitation light sources, and a low-cost field programmable gate array (FPGA) board (model: ZYNQ XC7Z020) as the master control unit. The fluorescence images captured by a complementary metal oxide semiconductor (CMOS) camera are processed by the YOLOv4-Tiny model embedded in FPGA to obtain the ultimate results of fish freshness. The circuit for the YOLOv4-Tiny model is optimized to make full use of FPGA resources and to increase computing efficiency. The performance of the device is evaluated by using grass carp fillets as the research object. The average accuracy of freshness detection reaches up to 97.10%. Moreover, the detection time of below 1 s per sample and the overall power consumption of 47.1 W (including 42.4 W light source power consumption) indicate that the device has good real-time performance and low power consumption. The research provides a potential tool for fish freshness evaluation in a low-cost and rapid manner. Full article

Biomass-derived carbon dots (CDs) are gaining much interest in recent times, as they provide a sustainable option with abundant availability, a low cost and tunable luminescence. Herein, we report a simple green synthesis method to produce highly fluorescent CDs from Eucalyptus globulus leaves using the one-pot hydrothermal approach. The fabricated CDs exhibit strong blue fluorescence with an excitation and emission maxima of 320 nm and 445 nm, respectively. The highest quantum yield (QY) obtained was 60.7%. With the reported optical properties and biocompatibility, CDs can be looked at as a promising candidate for potential biosensing applications. Moreover, we employed a life cycle assessment (LCA) cradle-to-gate approach to study the environmental impacts of the synthesis strategy used for the fabrication of CDs. The results point out that citric acid is the main hotspot in CD synthesis, regarding environmental impacts in most categories. This justifies the introduction of biomass, which reduces the amount of citric acid, thus leading to a more sustainable synthesis strategy for fabricating CDs. Full article

This paper presents a technique for high sensitivity measurement of singlet oxygen luminescence generated during photodynamic therapy (PDT) and ultraviolet (UV) irradiation on skin. The high measurement sensitivity is achieved by using a computational spectroscopy (CS) approach that provides improved photon detection efficiency compared to spectral filtering methodology. A solid-state InGaAs photodiode is used as the CS detector, which significantly reduces system cost and improves robustness compared to photomultiplier tubes. The spectral resolution enables high-accuracy determination and subtraction of photosensitizer fluorescence baseline without the need for time-gating. This allows for high sensitivity detection of singlet oxygen luminescence emission generated by continuous wave light sources, such as solar simulator sources and those commonly used in PDT clinics. The value of the technology is demonstrated during in vivo and ex vivo experiments that show the correlation of measured singlet oxygen with PDT treatment efficacy and the illumination intensity on the skin. These results demonstrate the potential use of the technology as a dosimeter to guide PDT treatment and as an analytical tool supporting the development of improved sunscreen products for skin cancer prevention. Full article

Saxitoxin (STX) causes high toxicity by blocking voltage-gated sodium channels, and it poses a major threat to marine ecosystems and human health worldwide. Our work evaluated the neurotoxicity and chronic toxicology of STX to Caenorhabditis elegans by an analysis of lifespan, brood size, growth ability, reactive oxygen species (ROS) and adenosine triphosphate (ATP) levels, and the overexpression of green fluorescent protein (GFP). After exposure to a series of concentrations of STX for 24 h, worms showed paralysis symptoms and fully recovered within 6 h; less than 5% of worms died at the highest concentration of 1000 ng/mL for first larval stage (L1) worms and 10,000 ng/mL for fourth larval stage (L4) worms. Declines in lifespan, productivity, and body size of C. elegans were observed under the stress of 1, 10, and 100 ng/mL STX, and the lifespan was shorter than that in controls. With STX exposure, the productivity declined by 32–49%; the body size, including body length and body area, declined by 13–18% and 25–27%, respectively. The levels of ROS exhibited a gradual increase over time, accompanied by a positive concentration effect of STX resulting in 1.14–1.86 times higher levels compared to the control group in L4 worms. Conversely, no statistically significant differences were observed between L1 worms. Finally, after exposure to STX for 48 h, ATP levels and GFP expression in C. elegans showed a significant dose-dependent increase. Our study reports the first evidence that STX is not lethal but imposes substantial oxidative stress on C. elegans, with a dose-responsive relationship. Our results indicated that C. elegans is an ideal model to further study the mechanisms underlying the fitness of organisms under the stress caused by paralytic shellfish toxins including STX. Full article

This study aims to investigate the optical properties of multiple neodymium-doped gadolinium compounds as a means to examine their eligibility as optical probes for fluorescence imaging. GdVO${}_4$, GdPO${}_4$, GdAlO${}_3$, Gd${}_2$SiO${}_5$ and Gd${}_3$Ga${}_5$O${}_{12}$ (GGG) samples were synthesized through solid-state reactions with varying neodymium doping levels to compare their optical properties in great detail. The optimal doping concentration was generally found to be approximately 2%. Furthermore, the luminescence lifetime, which is a valuable parameter for time-gated imaging, was determined to range from 276 down to 14 µs for the highest doping concentrations, resulting from energy transfer and migration assisted decay. Full article

Over the last few years, the development of fluorescent probes has received considerable attention. Fluorescence signaling allows noninvasive and harmless real-time imaging with great spectral resolution in living objects, which is extremely useful for modern biomedical applications. This review presents the basic photophysical principles and strategies for the rational design of fluorescent probes as visualization agents in medical diagnosis and drug delivery systems. Common photophysical phenomena, such as Intramolecular Charge Transfer (ICT), Twisted Intramolecular Charge Transfer (TICT), Photoinduced Electron Transfer (PET), Excited-State Intramolecular Proton Transfer (ESIPT), Fluorescent Resonance Energy Transfer (FRET), and Aggregation-Induced Emission (AIE), are described as platforms for fluorescence sensing and imaging in vivo and in vitro. The presented examples are focused on the visualization of pH, biologically important cations and anions, reactive oxygen species (ROS), viscosity, biomolecules, and enzymes that find application for diagnostic purposes. The general strategies regarding fluorescence probes as molecular logic devices and fluorescence–drug conjugates for theranostic and drug delivery systems are discussed. This work could be of help for researchers working in the field of fluorescence sensing compounds, molecular logic gates, and drug delivery. Full article

Free-running InGaAs/InP single-photon avalanche photodiodes (SPADs) typically operate in the active-quenching mode, facing the problems of long dead time and large timing jitter. In this paper, we demonstrate a 1-GHz gated InGaAs/InP SPAD with the sinusoidal gating signals asynchronous to the incident pulsed laser, enabling free-running single-photon detection. The photon-induced avalanche signals are quenched within 1 ns, efficiently reducing the SPAD’s dead time and achieving a count rate of up to 500 Mcount/s. However, the timing jitter is measured to be ~168 ps, much larger than that of the SPAD with synchronous gates. We adjust the delay between the gating signals and the incident pulsed laser to simulate the random arrival of the photons, and record the timing jitter, respectively, to figure out the cause of the jitter deterioration. In addition, the effects of the incident laser power and working temperature of the APD on the time resolution have been investigated, broadening the applications of the GHz gated free-running SPAD in laser ranging and imaging, fluorescence spectroscopy detection and optical time-domain reflectometry. Full article

Raman spectroscopy, a non-destructive reference technique, is used in heritage science to directly identify materials like pigments, minerals, or binding media. However, depending on the material, the laser source can induce a strong fluorescence signal that may mask the Raman signal during spectral detection. This photo-induced effect can prevent the detection of a Raman peak. A pulsed Raman spectroscopy, using a time-gated detection and pulsed laser, is proven capable of rejecting the fluorescence background and working with the environmental light, which makes Raman spectroscopy more adapted for in situ applications. In this paper, we investigated how an ns pulsed laser can be an excitation source of Raman spectroscopy by focusing on different parameters of laser excitation and collection. With proper implementation, this pulsed Raman technique can be used for cultural heritage with an ns pulsed laser for the first time. Full article

Time-gated fluorescence lifetime imaging microscopy with the o-BMVC fluorescent probe provides a visualizing method for the study of exogenous G-quadruplexes (G4s) in live cancer cells. Previously, imaging results showed that the parallel G4s are accumulated and that nonparallel G4s are not detected in the lysosomes of CL1-0 live cells. In this work, the detection of the G4 signals from exogenous GTERT-d(FN) G4s in the lysosomes may involve a structural change in live cells from intramolecular nonparallel G4s to intermolecular parallel G4s. Moreover, the detection of the G4 signals in the lysosomes after the 48 h incubation of HT23 G4s with CL1-0 live cells indicates the occurrence of structural conversion from the nonparallel G4s to the parallel G4s of HT23 in the live cells. In addition, the detection of much stronger G4 signals from ss-GTERT-d(FN) than ss-HT23 in the lysosomes of CL1-0 live cells may be explained by the quick formation of the intermolecular parallel G4s of ss-GTERT-d(FN) and the degradation of ss-HT23 before its intramolecular parallel G4 formation. This work provides a new approach to studying G4-lysosome interactions in live cells. Full article