Modulation Of Fluorescence Intensity Research Articles

Utility of native emission quenching of erythrosine B for the determination of diltiazem in different dosage forms.

This study introduces a practical and cost-effective method for tracking diltiazem (DLZ) analytically. It utilizes a fluorimetric approach that relies on the modulation of fluorescence intensity of a dye called erythrosine B. Through a one-pot experiment performed in an acidic environment, a complex is rapidly formed between DLZ and erythrosine B. By observing the decrease in erythrosine B emission, a linear calibration plot is established, enabling the detection and quantification of DLZ concentrations ranging from 40 to 850 ng/ml. The estimated limits of detection and quantitation were 10.5 and 32.1ng/ml, respectively. The variables affecting the DLZ-dye complex system were carefully adjusted. The validity of the approach was confirmed through a thorough evaluation based on the criteria set by ICH guidelines. The accuracy and precision of the methodology were evaluated, and the standard deviation and relative standard deviation were below 2. The strategy was successfully employed to analyze DLZ in tablets and capsules, and no significant variation between the proposed and reported methods as the values of the estimated t-test and F-test at five determinations were below 2.306 and 6.338, respectively. Notably, the method adheres to the principle of green chemistry by utilizing distilled water as the dispersing medium.

Multifaceted Cargo Recruitment and Release from Artificial Membraneless Organelles.

Liquid-liquid phase separation (LLPS) drives membraneless organelles (MLOs) formation for organizing biomolecules. Artificial MLOs (AMLOs) have been constructed mostly via the LLPS of engineered proteins capable of regulating limited types of biomolecules. Here, leveraging a minimalist AMLO, driven by LLPS of polymer-oligopeptide hybrids, enrichment, recruitment, and release of multifaceted cargoes are quantitatively shown, including small fluorescent molecules, fluorophore-containing macromolecules, proteins, DNAs, and RNAs. Cargoes show up to 105 -fold enrichment, whilst recruitment and release are triggered by variations of temperature, pH, and/or ionic strength. Also, the first efficacious, rapid, and reversible control of aggregation-induced emission with over 30 folds of modulation of overall fluorescence intensity is achieved, by intensifying the aggregation of luminogens in AMLO. The AMLO is a simple yet versatile platform for potential drug delivery and biosensor applications.

Glutathione-S-transferase Fusion Protein Nanosensor.

Fusion protein tags are widely used to capture and track proteins in research and industrial bioreactor processes. Quantifying fusion-tagged proteins normally requires several purification steps coupled with classical protein assays. Here, we developed a broadly applicable nanosensor platform that quantifies glutathione-S-transferase (GST) fusion proteins in real-time. We synthesized a glutathione-DNA-carbon nanotube system to investigate glutathione-GST interactions via semiconducting single-walled carbon nanotube (SWCNT) photoluminescence. We found that SWCNT fluorescence wavelength and intensity modulation occurred specifically in response to GST and GST-fusions. The sensor response was dependent on SWCNT structure, wherein mod(n - m, 3) = 1 nanotube wavelength and intensity responses correlated with nanotube diameter distinctly from mod(n - m, 3) = 2 SWCNT responses. We also found broad functionality of this sensor to diverse GST-tagged proteins. This work comprises the first label-free optical sensor for GST and has implications for the assessment of protein expression in situ, including in imaging and industrial bioreactor settings.

Fluorescence intensity modulation of CdSe/ZnS quantum dots assesses reactive oxygen species during chemotherapy and radiotherapy for cancer cells.

Quantum dots (QDs) are semiconductor nanoparticles ranging in size from 2 to 10 nm. QDs are increasingly being developed for biomedical imaging, targeted drug delivery and green energy technology. These have led to much research on QD interactions with various physical, chemical and biological systems. For biological systems, research has focused on the biocompatibility/cytotoxicity of QDs in the context of imaging/therapy. However, there is a paucity of work on how biological systems and bioactive molecules might be used to alter the optoelectronic properties of QDs. Here, it is shown that these properties can be altered by reactive oxygen species (ROS) from chemotherapeutic media and biological cells following controlled changes in cellular activities. Using CdSe/ZnS core-shell QDs, spectroscopic analysis of optically excited QDs with HL60, K562 and T98G cancer cell lines is performed. Our results show statistically significant (P < 0.0001) modulation of the fluorescence emission spectra of the QDs due to the ROS produced by common chemotherapeutic drugs, daunorubicin and doxorubicin and by cells following chemotherapy/radiotherapy. This optical modulation, in addition to assessing ROS generation, will possibly enhance applications of QDs in simultaneous diagnostic imaging and nanoparticle-mediated drug delivery as well as simultaneous ROS assessment and radiosensitization for improved outcomes in cancer treatments. Reactive molecular species produced by biological cells and chemotherapeutic drugs can create electric fields that alter the photophysical properties of QDs, and this can be used for concurrent monitoring of cellular activities, while inducing changes in those cellular activities.

High-contrast switching and high-efficiency extracting for spontaneous emission based on tunable gap surface plasmon

Controlling spontaneous emission at optical scale lies in the heart of ultracompact quantum photonic devices, such as on-chip single photon sources, nanolasers and nanophotonic detectors. However, achiving a large modulation of fluorescence intensity and guiding the emitted photons into low-loss nanophotonic structures remain rather challenging issue. Here, using the liquid crystal-tuned gap surface plasmon, we theoretically demonstrate both a high-contrast switching of the spontaneous emission and high-efficiency extraction of the photons with a specially-designed tunable surface plasmon nanostructures. Through varying the refractive index of liquid crystal, the local electromagnetic field of the gap surface plasmon can be greatly modulated, thereby leading to the swithching of the spontaneous emission of the emitter placed at the nanoscale gap. By optimizing the material and geometrical parameters, the total decay rate can be changed from 103γ0 to 8750γ0, [γ0 is the spontaneous emission rate in vacuum] with the contrast ratio of 85. Further more, in the design also enables propagation of the emitted photons along the low-loss phase-matched nanofibers with a collection efficiency of more than 40%. The proposal provides a novel mechanism for simultaneously switching and extracting the spontaneous emitted photons in hybrid photonic nanostructures, propelling the implementation in on-chip tunable quantum devices.

Integration of an oxygen sensor into a polydymethylsiloxane hepatic culture device for two-dimensional gradient characterization

This paper presents a new hepatic culture device, in which oxygen concentration gradient formed by cellular respiration can be visualized as a two-dimensional optical image. An oxygen sensor layer containing Pt (II) octaethylporphyrin (PtOEP) was embedded at the bottom of the device, allowing visualization of oxygen concentration distribution over the cell culture area. Oxygen concentration was determined by analyzing the modulation of fluorescence intensity emitted by the sensor, which is negatively correlated to the concentration. The results in HepG2 cell culture showed that the oxygen gradient and local oxygen partial pressure could be regulated by controlling the perfusion flow rate in the device. The local and physiological oxygen gradient comparable to the in vivo liver lobules can successfully be reproduced in the device. Therefore, periportal and perivenous hepatic like-zones were successfully realized in vitro. The present device would be helpful in the development of novel in vitro methods recapitulating the liver physiology for investigation of hepatic zonation and drug screening system recapitulating the liver physiology.

An Optical Sensor Array Discriminates Syrups and Honeys

We report a cross-reactive sensor array, combining a two-component probe system based upon three viologen substituted boronic acids and a poly(aryleneethynylene) (PAE) and an additional number of simple PAEs alone. This combined system discriminates 27 different honeys in aqueous solution, according to patterns in fluorescence intensity modulation, using linear discriminant analysis for data processing. The fluorescence turn-on array detects saccharide composition, while the PAEs discriminate trace colored components in the honeys.

Organic nanoparticles of an extended π-conjugated styryl dye: Modulation of fluorescence peak energy and intensity in the near-infrared (NIR) region

Abstract Tuning the near-infrared (NIR) emission of organic nanomaterials is of great interest due to possible use in biosensing and imaging based on the biological window ranged in 650–900 nm. In this study, we report NIR fluorescent organic nanoparticles prepared on the basis of ion association between an extended π-conjugated Styryl-9 cation and tetrakis(4-fluorophenyl)borate (TFPB) anion. Binding of Styryl-9 with TFPB results in the prominent appearance of an absorption band that can be assigned to a non-fluorescent H-aggregate, which is revealed by simultaneous deconvolution of both UV–vis absorption and fluorescence excitation spectra, and supported by quantum chemical calculations. Emission is observed in the NIR region from the nanoparticles, but is reduced by the presence of the H-aggregates. To mitigate the H-aggregation occurred inside the nanoparticle, photochemically-inert, bulky tetraphenylphosphonium cation (co-cation) is concurrently incorporated (co-doped), resulting in the disappearance of the relevant H-aggregates and thereby yielding controllable fluorescence peak energy and intensity in the NIR region. We believe that such methodology for synthesizing ion-based organic nanoparticles with tunable emission will open an avenue in the development of new types of NIR fluorescent nanomaterials.

Highly efficient modulation of FRET in an orthogonally arranged BODIPY-DTE dyad.

The photoswitchable boron-dipyrromethene–dithienylethene molecular dyad is introduced as a prototype for the efficient fluorescence intensity modulation on the molecular level. The functionality of the system is based on the photochromism of the dithienylethene, which facilitates an efficient on- and off-switching of a Förster-type intramolecular energy transfer between the photoexcited BODIPY donor and the dithienylethene acceptor moiety. The switching behavior and dynamics of the molecular dyad are monitored by steady state and time-resolved spectroscopic methods. A quenching efficiency of up to 96% in the off-state is observed and explained by a drastically accelerated decay of the boron-dipyrromethene excited state due to the efficient energy transfer despite the orthogonal arrangement of donor and acceptor. An energy transfer time orders of magnitude shorter than the lifetime of the boron-dipyrromethene in the open state is determined.

An Automated High-Throughput Cell-Based Multiplexed Flow Cytometry Assay to Identify Novel Compounds to Target Candida albicans Virulence-Related Proteins

Although three major classes of systemic antifungal agents are clinically available, each is characterized by important limitations. Thus, there has been considerable ongoing effort to develop novel and repurposed agents for the therapy of invasive fungal infections. In an effort to address these needs, we developed a novel high-throughput, multiplexed screening method that utilizes small molecules to probe candidate drug targets in the opportunistic fungal pathogen Candida albicans. This method is amenable to high-throughput automated screening and is based upon detection of changes in GFP levels of individually tagged target proteins. We first selected four GFP-tagged membrane-bound proteins associated with virulence or antifungal drug resistance in C. albicans. We demonstrated proof-of-principle that modulation of fluorescence intensity can be used to assay the expression of specific GFP-tagged target proteins to inhibitors (and inducers), and this change is measurable within the HyperCyt automated flow cytometry sampling system. Next, we generated a multiplex of differentially color-coded C. albicans strains bearing C-terminal GFP-tags of each gene encoding candidate drug targets incubated in the presence of small molecules from the Prestwick Chemical Library in 384-well microtiter plate format. Following incubation, cells were sampled through the HyperCyt system and modulation of protein levels, as indicated by changes in GFP-levels of each strain, was used to identify compounds of interest. The hit rate for both inducers and inhibitors identified in the primary screen did not exceed 1% of the total number of compounds in the small-molecule library that was probed, as would be expected from a robust target-specific, high-throughput screening campaign. Secondary assays for virulence characteristics based on null mutant strains were then used to further validate specificity. In all, this study presents a method for the identification and verification of new antifungal drugs targeted to fungal virulence proteins using C. albicans as a model fungal pathogen.

Manipulating nanoscale features on the surface of dye-loaded microbubbles to increase their ultrasound-modulated fluorescence output.

The nanoscale surface features of lipid-coated microbubbles can dramatically affect how the lipids interact with one another as the microbubble diameter expands and contracts under the influence of ultrasound. During microbubble manufacturing, the different lipid shell species naturally partition forming concentrated lipid islands. In this study the dynamics of how these nanoscale islands accommodate the expansion of the microbubbles are monitored by measuring the fluorescence intensity changes that occur as self-quenching lipophilic dye molecules embedded in the lipid layer change their distance from one another. It was found that when the dye molecules were concentrated in islands, less than 5% of the microbubbles displayed measurable fluorescence intensity modulation indicating the islands were not able to expand sufficiently for the dye molecules to separate from one another. When the microbubbles were heated and cooled rapidly through the lipid transition temperature the islands were melted creating an even distribution of dye about the surface. This resulted in over 50% of the microbubbles displaying the fluorescence-modulated signal indicating that the dye molecules could now separate sufficiently to change their self-quenching efficiency. The separation of the surface lipids in these different formations has significant implications for microbubble development as ultrasound and optical contrast agents.

Electrofluorochromism in π-conjugated ionic liquid crystals

Materials in which photoluminescence is modulated by redox processes are known as electrofluorochromic. Intrinsically switchable fluorophores, incorporating both redox and fluorescent moieties, could be ideal electrofluorochromic materials if they possess high fluorescence quantum yields in at least one of their redox states. Fluorescent liquid crystals with redox active centres could combine the above requirements with the advantage to work in bulk anisotropic phases. However, electrofluorochromic liquid crystals have not been reported yet because their synthesis is challenging due to aggregation-caused fluorescent quenching. Here we show the first examples of electrofluorochromic π-conjugated ionic liquid crystals based on thienoviologens. These ordered materials, combining ionic and electronic functions, are highly fluorescence in the bulk state (quantum yield>60%). Their direct electrochemical reduction leads to fast and reversible bulk electrofluorochromic response in both columnar and smectic phases allowing for fluorescence intensity modulation and colour tuning.

Angular modulation of single-molecule fluorescence by gold nanoparticles on DNA origami templates

Abstract We study the angular fluorescence intensity modulation of a single dye positioned near a spherical gold nanoparticle, induced by rotation of linearly polarized excitation light. Accurate positioning and alignment of nanoparticle and fluorophore with respect to each other and the incoming electric field is achieved by a three-dimensional, self-assembled DNA origami. An intensity map is obtained for a fixed distance and two different nanoparticle diameters, revealing polarization-dependent enhancement and quenching of fluorescence intensity in good agreement to numerical simulations.

Fluorescence Intensity Modulation in Photochromic Conjugated Polymer Systems

AbstractThe ability to modulate fluorescence intensity with a light signal enables a variety of applications based on fluorescence imaging. One approach to fluorescence photomodulation involves using a photochromic moiety that responds to a light signal in conjunction with a nearby fluorophore. We employ conjugated polymers based on poly(p‐phenylene vinylene) (PPV) as the fluorophore in photochrome‐fluorophore systems for fluorescence modulation. Advantages of using conjugated polymers for this purpose include their intrinsic energy migration processes that enable amplified fluorescence quenching as well as their processability. Here we present examples of PPV‐based photomodulation systems that employ photochromic dyes from three common photochromic families: azobenzenes, spironaphthoxazines, and diarylethenes. In all cases we observe reversible fluorescence quenching due to fluorescence resonance energy transfer to the photogenerated form of the photochrome. Examples of the photomodulation of photochromic PPV systems in organic solution, polymer films, and conjugated polymer nanoparticles are presented.

Phospholipid/carbocyanine dye-shelled microbubbles as ultrasound-modulated fluorescent contrast agents

Fluorescent microbubbles have been fabricated with the capacity to have their emission modulated by ultrasound. These contrast agent particles could potentially be used in the future to extract fluorescence modulation from a strong light background to increase imaging depth and resolution in scattering media. Fluorescence intensity modulation was demonstrated at the ultrasound driving frequency.

Other origins for the fluorescence modulation of single dye molecules in open-circuit and short-circuit devices

Fluorescence intensity modulation of single Atto647N dye molecules in a short-circuit device and a defective device, caused by damaging an open-circuit device, is due to a variation in the excitation light focus as a result of the formation of an alternating electric current.

Luminescence switching of CdTe quantum dots in presence of water-soluble spironaphthoxazine

The typical reaction of spironaphthoxazine is the conversion between the non-polar spiro form and polar merocyanine form. Quantum dots (QDs) have unique optical properties and several advantages over conventional organic dyes. In this work, the CdTe QDs containing a water-soluble spironaphthoxazine was used as the optical probe photoisomerization between spiro and merocyanine. The initial bright fluorescence of CdTe QDs was effectively quenched by water-soluble spironaphthoxazine using UV irradiation. Reversible modulation of fluorescence intensity of CdTe QDs was achieved using alternating irradiation with UV and visible light. The possible mechanism is discussed.

Modulation of a fluorescence switch of nanofiber mats containing photochromic spironaphthoxazine and D-π-A charge transfer dye

Photoswitchable poly(methyl methacrylate) (PMMA) nanofiber mats containing spironaphthoxazine (SPO)/electron donor-π-acceptor (D-π-A) type fluorescent dye (TCF) were prepared by electrospinning. The photoregulated fluorescence switching behaviors of SPO/TCF blended solution and PMMA nanofiber mats containing SPO/TCF were also studied. Not only SPO/TCF blended solution but also PMMA nanofiber mats containing SPO/TCF showed reversible modulation of fluorescence intensity using alternating irradiation with UV and visible light.

Switching properties of fluorescent photochromic poly(methyl methacrylate) with spironaphthoxazine and D-π-A type pyran-based fluorescent dye

Fluorescent photochromic poly(methyl methacrylate) (PMMA) with spironaphthoxazine (SPO) and D-π-A type pyran-base fluorescent dye as a fluorophore was synthesized by typical free radical copolymerization. The poly(MMA- co-SPO- co-fluorophore) in both solution and solid film exhibited excellent photoregulated fluorescence switching behavior and reversible modulation of fluorescence intensity using alternating irradiation with UV and visible light. The poly(MMA- co-SPO- co-fluorophore) also showed viscosity and conductivity switching behaviors along with photoresponse.

Multiple switching behaviors of poly( N-isopropylacrylamide) hydrogel with spironaphthoxazine and D-π-A type dye

A multi-switchable poly(NIPAM- co-SPO- co-D-π-A dye) hydrogel with a photochromic spironaphthoxazine and an electron donor-π-conjugated-electron acceptor (D-π-A) type dye was prepared by typical radical copolymerization. The low critical solution temperature (LCST) behavior was investigated by UV–vis spectroscopy, which allows the measurement of the phase transition from 20 to 40 °C in aqueous solution. The fluorescence intensity of the poly(NIPAM- co-SPO- co-D-π-A dye) was temperature-dependent. Reversible modulation of fluorescence intensity was achieved using alternating irradiation with UV and visible light. Prepared polymer hydrogel also exhibited spectra change when not only used Cu 2+ cation but also an acid unit.