Time-resolved Fluorescence Spectroscopy Research Articles

High Fluorescence of Phytochromes Does Not Require Chromophore Protonation

Fluorescing proteins emitting in the near-infrared region are of high importance in various fields of biomedicine and applied life sciences. Promising candidates are phytochromes that can be engineered to a small size and genetically attached to a target system for in vivo monitoring. Here, we have investigated two of these minimal single-domain phytochromes, miRFP670nano3 and miRFP718nano, aiming at a better understanding of the structural parameters that control the fluorescence properties of the covalently bound biliverdin (BV) chromophore. On the basis of resonance Raman and time-resolved fluorescence spectroscopy, it is shown that in both proteins, BV is deprotonated at one of the inner pyrrole rings (B or C). This protonation pattern, which is unusual for tetrapyrroles in proteins, implies an equilibrium between a B- and C-protonated tautomer. The dynamics of the equilibrium are slow compared to the fluorescence lifetime in miRFP670nano3 but much faster in miRFP718nano, both in the ground and excited states. The different rates of proton exchange are most likely due to the different structural dynamics of the more rigid and more flexible chromophore in miRFP670nano3 and miRFP718nano, respectively. We suggest that these structural properties account for the quite different fluorescent quantum yields of both proteins.

Exploring Unusual Confined Chemistry: Excited-State Proton Transfer Reaction in Supported Liquid Membrane with Deep Eutectic Solvents.

Supported liquid membrane (SLM) incorporating ionic liquids (ILs) or deep eutectic solvents (DESs) offers a promising method for ion and (bio)chemical separations and CO2 capture. However, a molecular understanding of whether chemical reactions occur in these confined media is crucial. We report excited-state proton transfer (ESPT) reaction of a photoacid, HPTS, in various DES-based SLMs (pore size ∼280 nm) using steady-state and time-resolved fluorescence spectroscopy. Our findings reveal that, while the ESPT is unfavorable in bulk DESs, it occurs substantially in SLM-containing DESs. Time-resolved area normalized emission spectra (TRANES) show that ESPT time ranges from 2.6 to 7.5 ns and is greatly affected by changes in DES constituents. The results suggest that HPTS interacts with ordered DES structures formed by long-range interfacial effects in membrane pores, making it suitable for ESPT. Furthermore, it is found that the dynamics of solvent relaxation in confined DESs are significantly slower than in bulk liquids. This observation, together with a large red-edge excitation shift, supports the impact of long-range interfacial effects on the DES structure inside membrane pores. Given the task-specific properties of DESs, the incorporation of these solvents into SLM pores can be a useful strategy for investigating new chemical processes.

The Interaction Of Homodimer Styrylcyanine Dyes With Sodium Dodecyl Sulfate And Triton X-100.

Solubilization of the styrylcyanine dye Sbt ((E)-2-(4-(dimethylamino)styryl)-3-methylbenzo[d]thiazol-3-ium iodide) and its homodimers Dbt-5 and Dbt-10 in aqueous solution of sodium dodecyl sulfate and Triton X-100 has been studied by steady state and picosecond time-resolved fluorescence spectroscopy. At low concentration of sodium dodecyl sulfate in solution, between Sbt, Dbt-5 dyes molecules and surfactant ion pairs are formed followed by the formation non-luminescent H-aggregates. The nature of the interaction between molecules of dyes and surfactants has been revealed. The binding constants Ks of the dyes to the surfactants, free energy changes (ΔG0), the number of dye molecules (n) included in a single micelle and photophysical parameters have been determined. The degree of solubilization of dyes in micellar solution of Triton X-100 is higher as compared to sodium dodecyl sulfate and depends on the molecular weight and size of both dye molecules and micelles.

Integrating synthetic hydrogel nanoparticles with carbon dots for selective detection of hemoglobin

Synthetic hydrogel nanoparticles (NPs) display high affinity to biomacromolecules target and have significant potential as protein capture agents for medical and biotechnological applications. In this study, we proposed a strategy to modify Carbon dots (CDs) with engineered NPs to address their detection specificity in complex biological sensing matrices. A library of NPs tagged CDs (NPs@CDs) incorporating hydrophobic and carboxylate groups that bound with high affinity to hemoglobin (Hb) was prepared by precipitation polymerization and screened based on the fluorescence quenching effect. NPs and CDs were integrated through noncovalent interaction, which was confirmed by Transmission electron microscopy (TEM), Scanning electron microscopy (SEM), Dynamic Light Scattering (DLS) and Fourier transform infrared spectroscopy analyses. The specificity of NPs@CDs was prominently attributed to the unique affinity between the screened NPs and Hb, compared with the contrast fluorescence quenching study by corresponding CDs to structural analogues and coexistence substances of Hb. Moreover, the Hb-induced fluorescence quenching process was dominated by a synergistic effect of internal filter effect (IFE) and static quenching proved by UV–vis absorption spectrometry and time-resolved fluorescence spectrometry. The screened NPs@CDs showed strong recognition capability to Hb, and the fluorescence quenching rate was about 97 % which was 3.2 times that of CDs. A biosensing platform based on NPs@CDs was constructed, and the fluorescence signal of the nanoprobe decreased linearly as the concentration of Hb over the range of 0.2–80 μM L−1 with a detection limit of 7.5 nM L−1. The synergy between CDs and NPs resulted in high-speed real-time detection capability, high sensitivity, considerable specificity, low interference and good stability of this nanoprobe.

Unified Role of the 145th Residue on the Fluorescence Lifetime of Fluorescent Proteins from the Jellyfish Aequorea victoria.

Finding a unified fluorescence mechanism is essential to develop and utilize fluorescent proteins appropriately. Here, we report the unified role of the 145th residue on the fluorescence efficiency of fluorescent proteins developed from the jellyfish Aequorea victoria by demonstrating the difference and similarity between two representative fluorescent proteins, enhanced green fluorescent protein (eGFP), and enhanced yellow fluorescent protein (eYFP). We determined the fluorescence lifetimes of the 19 different Y145 mutants of eGFP and eYFP by picosecond time-resolved fluorescence spectroscopy. We found that the effect of the 145th mutation on the fluorescence lifetime is significant for eYFP but moderate for eGFP. We compared known crystal structures to clarify the observed difference between eGFP and eYFP. As a result, we conclude that the efficiency of the steric restriction of the chromophore motion by the 145th side chain is essentially the same for both eGFP and eYFP. Meanwhile, the restriction of the chromophore motion by hydrogen bonds is more pronounced for eGFP than for YFP. Balance of the steric effect and hydrogen bonding controls the lifetime of the Y145 mutants for eGFP and eYFP. Furthermore, the steric restriction is induced by the electrostatic effect; the different 145th residue induces a different electrostatic environment around the chromophore. The finding in this study reasonably explains the reported lifetimes of other fluorescent proteins and allows the prediction of the lifetime of unknown fluorescent proteins from jellyfish.

Insight into the complexation of uranium with Bacillus sp. dwc-2 by multi-spectroscopic approaches: FT-IR, TRLF and XAFS spectroscopies

The micromechanisms of the interactions between uranium and microorganisms, especially chemical bonds and bonding numbers between uranium and microbe are not yet clear. In this work, uranium coordination with functional groups in/on Bacillus sp. dwc-2 has been rationally explored by X-ray absorption fine structure (XAFS), Fourier transform infrared (FT-IR), time-resolved laser-induced fluorescence (TRLF) spectroscopies. The results indicate that under acidic condition, 66.8 % of O atoms in coordination shell of U(VI) equatorial plane are provided by organic phosphoryl groups via monodentate coordination, 18.8 % and 15.5 % are provided by water molecules and carboxyl groups via monodentate and bidentate coordination, respectively. While under alkaline condition, these proportions change to 53.1 %, 34.4 % and 22.0 %, respectively. Under neutral condition, U(VI) is mainly deposited by complexing with inorganic phosphates released by microbial cells. Under anaerobic condition, partial U(VI) can be reduced to U(IV) which primarily complex with phosphorus containing groups via monodentate and bidentate coordination, existing as amorphous solids or coordination polymers. This is the first exploration of the binding mechanism between uranium and microorganisms at a semi-quantitative level under different conditions. Our studies will further enhance the contribute to the understanding of coordination mechanism of uranium with microorganism, and demonstrate the potential bioremediation value of this bacterium.

Growth of anthracene microcrystals by the micro-space sublimation method and their photophysical properties

Anthracene and its derivatives are widely utilized in optoelectronic devices due to their unique properties. Generally, single-crystal structures can avoid non-radiative recombination, enhance carrier mobility, and ultimately improve device performance. In this work, anthracene microcrystals were prepared using the micro-space sublimation method. Through real-time in-situ observation, the crystallization dynamics of anthracene molecules were revealed. Unlike traditional vacuum evaporation deposition technique, the close proximity of the substrate to the source facilitates the self-assembly of anthracene molecules into an ordered crystal structure. Six peaks can be observed in the photoluminescence spectrum, corresponding to various lowest excited state decay processes. The fluorescence intensity at the peak of 423 nm decreases significantly with increasing temperature. The reason for this is the relatively high exciton binding energy, which makes excitons more stable and easier to form. The lattice vibrations induced by increased temperature were found to affect the transport and separation of excitons. Time-resolved fluorescence spectroscopy imaging revealed that a relatively uniform distribution of fluorescence lifetimes in the anthracene microcrystals, indicating high crystallization quality. This work provides valuable insights for controlling the morphology and investigating the photophysical properties of organic semiconductors.

Lentil Extract-Mediated Ag QD Synthesis: Predilection for Albumin Protein Interaction, Antibacterial Activity, and Its Cytotoxicity for Wi-38 and PC-3 Cell Lines.

Recent focus has been directed toward semiconductor nanocrystals owing to their unique physicochemical properties. Nevertheless, the synthesis and characterization of quantum dots (QDs) pose considerable challenges, limiting our understanding of their interactions within a biological environment. This research offers valuable insights into the environmentally friendly production of silver quantum dots (Ag QDs) using lentil extract and clarifies their distinct physicochemical characteristics, previously unexplored to our knowledge. These findings pave the path for potential practical applications. The investigation of the phytochemical-assisted Ag QDs' affinity for BSA demonstrated modest interactions, as shown by the enthalpy and entropy changes as well as the associated Gibbs free energy during their association. Steady-state and time-resolved fluorescence spectroscopy further demonstrated a transient effect involving dynamic quenching, predominantly driven by Forster resonance energy transfer. Additionally, the study highlights the potential broad-spectrum antibacterial activity of Ag QDs (<5 nm, a zeta potential of -3.04 mV), exhibiting a remarkable MIC value of 1 μg/mL against Gram-negative bacteria (E. coli) and 1.65 μg/mL against Gram-positive bacteria (S. aureus). They can readily enter cells and tissues due to their minuscule size and the right chemical environment. They cause intracellular pathway disruption, which leads to cell death. This outcome emphasizes the distinctive biocompatibility of the green-synthesized Ag QDs, which has been confirmed by their MTT assay-based cytotoxicity against the PC-3 and Wi-38 cell lines.

Dynamic spectroscopic and optical characterization and modeling of bovine serum albumin corona during interaction with N-hydroxysulfo-succinimide-covalently functionalized gold nanourchins.

In this study, bovine serum albumin (BSA) is used as a globular protein model to examine the conformational changes that occur during the interaction of BSA with N-hydroxysulfo-succinimide (sodium salt)-functionalized gold nanourchins (GNUs), for which dynamic spectroscopic techniques are employed. The results showed that the absorbance of phosphate-buffered saline-BSA at 278 nm decreased when a GNU was added to the solution due to adsorption, and it decreased further when the GNU was increased. The intensity and width of the peak of local surface plasmon resonance increased, indicating the effect of corona formation. Dynamic UV-vis spectroscopy and scattering revealed a nonlinear behavior of BSA-GNU interaction. The bioplasmonic solution resulted in higher transmission and scattering than the BSA solution. Fourier transform-near-infrared spectra exhibited several bands due to overtones and combinations of the amide group and different proportions of α-helix and β-sheet components in BSA before and after the addition of the GNU. Time-resolved fluorescence spectroscopy demonstrated an initial increase in blueshifted emission, followed by a redshifted quenching of two major peaks of Tyr and tryptophan (Trp). The binding and dissociation constants were determined as Kb = 2.17 × 1010 M-1 and Kd = 4.6 × 10-11, respectively, using the Stern-Volmer relation. Both the dynamic CMOS-based imaging and the cadmium sulfide sensors demonstrated a nonlinear response of bioplasmonic solution. By increasing the GNU, the resistance of the solution decreased in the order of A > S1 > S3, where S3 exhibited the highest initial transmission with a longer desorption time. MATLAB modeling showed 80% surface coverage by the protein in 15 s at 0.05M, equivalent to a thickness of 1.7 nm, which was in agreement with the value determined by using the Stokes-Einstein relation.

Cm(III) speciation in the presence of citrate from neutral to hyperalkaline conditions and the effect of calcium

We investigated the binary Cm-citrate system using time-resolved laser fluorescence spectroscopy (TRLFS), parallel factor analysis (PARAFAC), and quantum chemical calculations. Evidence collectively suggests the stepwise coordination and deprotonation of citrate alcohol groups in Cm-cit complexes with two bound citrate moieties upon increasing pH, which is supported by a bathochromic shift in emission spectra, an observed increase in lifetime measurements, and lower energy minima for citrate alcohol involvement versus hydrolysis of the Cm-citrate species. Our PARAFAC results agree with a 3-component model for the Cm-citrate system and offer pure component decompositions, yielding fraction species across the studied pH range that have a correlated slope = 1 as a function of pH. For the first time, evidence of ternary Ca-Cm-citrate complexes was revealed by TRLFS with increasing calcium concentration at fixed pHm. The formation of these ternary complexes was substantiated with density functional theory (DFT) calculations on simple model systems of the complexes. Shared citrate carboxylate groups between calcium and curium were proposed for all three ternary Ca-Cm-cit complexes based on DFT-determined Ca–O and Cm–O distances. Moreover, we found that the ternary complex with both alcohol groups deprotonated is most stable when it shares both two carboxylate and two alcohol groups between Ca and Cm. The presence of shared functional groups highlights the enhanced stability of these ternary complexes. Additional work is warranted to further constrain the stoichiometry, stability constants and dependence on ionic strength of these complexes for purposes of thermodynamic modeling of repository settings.

Self-Indicating Polymer Prodrug Nanoparticles for pH-Responsive Drug Delivery in Cancer Cells and Real-Time Monitoring of Drug Release.

Amphiphilic self-indicating and responsive polymer-based prodrugs have generated much interest as potential stimuli-responsive intelligent drug delivery systems (DDS) due to their ability to selectively deliver drugs to the cancer cells and to monitor real-time cellular uptake of the drug by imaging technique(s). In this direction, we have synthesized a new pH-responsive N-vinyl-2-pyrrolidone and coumarin-based fluorescent self-indicating polymeric prodrug (SIPD), poly(NVP)-b-poly(FPA.DOX-r-FPA-r-CA). This block copolymer prodrug self-assembled into stable micellar nanoparticles under physiological conditions that reduced undesirable drug leakage to normal cells but resulted in the release of the anticancer drug doxorubicin (DOX) in cancer cells because of acidic pH-induced cleavage of imine bonds between DOX and the copolymer. While the polymer was found to be highly biocompatible with both normal (HEK-293) cells and cancer (MCF-7) cells even at high concentrations by MTT assay, the polymer prodrug nanoparticles showed toxicity even higher than that of free DOX in cancer cells. Phase contrast microscopy also depicted the cytotoxic effects of the nanoparticles on cancer cells. The coumarin units present in the polymer served as a fluorescence resonance energy transfer (FRET) pair with the covalently attached DOX molecules, which was established by steady-state and time-resolved fluorescence spectroscopy. Furthermore, confocal microscopy results confirmed the FRET phenomenon, as the fluorescence intensity of coumarin in the micellar nanoparticles remained quenched initially in MCF-7 cells but recovered with time as the DOX molecules were released and gradually shifted toward the targeted nucleus. All of these studies implied that the synthesized prodrug nanoparticles may provide another viable option for delivering chemotherapeutic drugs into cancer cells with a capability of real-time monitoring of drug release.

Synthetic Protein Nanoparticles via Photoreactive Electrohydrodynamic Jetting.

Protein nanoparticles are an attractive class of materials for nanomedicine applications due to the intrinsic biocompatibility, biodegradability, and intrinsic functionality of their constituent proteins. Despite the clinical success of select protein nanoparticles, this class of nanocarriers remains understudied and underdeveloped compared to lipid and polymer nanoparticles due to challenges related to formulation optimization, large design space, and their structural complexity. In this work, a modular strategy for protein nanoparticle preparation based on the concept of photoreactive jetting is introduced. The process relies on continuous ultravioletirradiation during electrohydrodynamic (EHD) jetting of protein solutions that contain a homobifunctional photocrosslinker. Protein nanoparticles exhibit nanogel-like architectures comprised of proteins that are linked via synthetic moieties. Compared to conventional protein nanoparticles, this method reduces nanoparticle processing times to minutes, rather than hours to days. The inclusion of an emissive structural motif as the molecular scaffold of the photocrosslinker is used to study the supramolecular architecture of the stable nanoparticles via time-resolved fluorescence spectroscopy.

Investigating the interaction of uranium(VI) with diatoms and their bacterial community: A microscopic and spectroscopic study

Diatoms and bacteria play a vital role in investigating the ecological effects of heavy metals in the environment. Despite separate studies on metal interactions with diatoms and bacteria, there is a significant gap in research regarding heavy metal interactions within a diatom-bacterium system, which closely mirrors natural conditions. In this study, we aim to address this gap by examining the interaction of uranium(VI) (U(VI)) with Achnanthidium saprophilum freshwater diatoms and their natural bacterial community, primarily consisting of four successfully isolated bacterial strains (Acidovorax facilis, Agrobacterium fabrum, Brevundimonas mediterranea, and Pseudomonas peli) from the diatom culture. Uranium (U) bio-association experiments were performed both on the xenic A. saprophilum culture and on the four bacterial isolates. Scanning electron microscopy and transmission electron microscopy coupled with spectrum imaging analysis based on energy-dispersive X-ray spectroscopy revealed a clear co-localization of U and phosphorus both on the surface and inside A. saprophilum diatoms and the associated bacterial cells. Time-resolved laser-induced fluorescence spectroscopy with parallel factor analysis identified similar U(VI) binding motifs both on A. saprophilum diatoms and the four bacterial isolates. This is the first work providing valuable microscopic and spectroscopic data on U localization and speciation within a diatom-bacterium system, demonstrating the contribution of the co-occurring bacteria to the overall interaction with U, a factor non-negligible for future modeling and assessment of radiological effects on living microorganisms.

Application of Steady-State and Time-Resolved Fluorescence Spectroscopy in Identification of Bee Products

In this study, steady-state and time-resolved fluorescence spectroscopy techniques have been applied to determine fluorescence characteristics and fluorescence decay kinetics parameters (fluorescence lifetimes and their amplitudes) of available on the Polish market bee products, including several nectar honeys, royal jelly, bee bread in honey and in liquid artificial honey. The fluorescence properties of the tested bee products arise from the presence of a unique composition of aromatic amino acids, vitamins, phenolic compounds and Maillard reaction products. In the 300–550 nm region of the emission spectra (excited at 280 nm), each of the tested bee products exhibited (showed) a specific and distinctive vibronic structure, which was not observed in the spectrum of artificial honey. Quantitative and qualitative composition as well as specific interactions between fluorescent constituents determine the specific fluorescence characteristics of a given bee product providing a unique fingerprint that can be used in the identification of bee products of different botanical origin. Combination of stationary and time-resolved fluorescence techniques seems to be a promising approach in the identification, authentication and quality control of bee products to verify their health-beneficial properties.

(Invited) Comprehensive Analysis of Exciton Diffusion with Time-Resolved Fluorescence Spectroscopy, Anisotropy and Imaging

Excitons produced in materials play a crucial role in photo-energy conversion such as photovoltaics and electroluminescence devices. The temporal and spatial diffusion of excitons is a key factor for dominating the fundamental performance of optoelectronic materials. In this context, analysis of the exciton diffusion dynamics is an important issue for obtaining the rational design of the materials. Excitons diffusing in materials can be characterized by several parameters such as the energy level, orientation of the transition dipole moment and spatial distribution, and the detection of these properties is required for the comprehensive analysis. In the present study, we show time-resolved fluorescence methods for detecting the above relevant properties of excitons.First, time-resolved fluorescence spectroscopy is a basic technique for analyzing the exciton diffusion dynamics. Fluorescence intensity is detected as functions of observation wavelength and time, and the spectral intensity and shift of fluorescence bands provide information on the population of excitons and their energy level. For example, dynamic Stokes shift indicates downhill energy migration and excitons are trapped by the defective site in molecular aggregates. Second, fluorescence anisotropy can be a good indicator for tracking the exciton diffusion in amorphous materials. Fluorescence anisotropy is sensitive to orientation change in transition dipole moments (TDMs) between absorption and fluorescence. In systems where the relative orientation of adjacent molecules is different from each other, such as amorphous solids, the exciton diffusion is accompanied with the change in TDM. Thus, appropriate modeling of molecular alignment enables quantitative estimation of the exciton diffusion coefficient and diffusion length from the anisotropy signal. Finally, time-resolved imaging is a combination of time-resolved spectroscopy and super-resolution microscopy, and an emergent technique for visualizing real-space propagation of excitons in materials. The diffraction-limited excitation beam produces excitons in several hundreds of nanometers and the subsequent exciton diffusion is evaluated by the spatial width of the fluorescence spot. The temporal broadening of the fluorescence spot directly reflects the exciton distribution and we can intuitively obtain the diffusion coefficient and length. Unlike the conventional methods for bulk materials, the advantage of the time-resolved imaging is the applicability to materials with nano- and meso-scale heterogeneous structure and it enables site-selective evaluation of the exciton transport capability. In the conference site, we will also show the application of the above methods to supramolecular polymers and thermally activated delayed fluorescence materials. Figure 1

Self-Reporting Therapeutic Protein Nanoparticles.

We present a modular strategy to synthesize nanoparticle sensors equipped with dithiomaleimide-based, fluorescent molecular reporters capable of discerning minute changes in interparticle chemical environments based on fluorescence lifetime analysis. Three types of nanoparticles were synthesized with the aid of tailor-made molecular reporters, and it was found that protein nanoparticles exhibited greater sensitivity to changes in the core environment than polymer nanogels and block copolymer micelles. Encapsulation of the hydrophobic small-molecule drug paclitaxel (PTX) in self-reporting protein nanoparticles induced characteristic changes in fluorescence lifetime profiles, detected via time-resolved fluorescence spectroscopy. Depending on the mode of drug encapsulation, self-reporting protein nanoparticles revealed pronounced differences in their fluorescence lifetime signatures, which correlated with burst- vs diffusion-controlled release profiles observed in previous reports. Self-reporting nanoparticles, such as the ones developed here, will be critical for unraveling nanoparticle stability and nanoparticle-drug interactions, informing the future development of rationally engineered nanoparticle-based drug carriers.

6-(6-Methyl-1,2,4,5-Tetrazine-3-yl)-2,2'-Bipyridine: A N-Donor Ligand for the Separation of Lanthanides(III) and Actinides(III).

Here, we report the synthesis of the 6-(6-methyl-1,2,4,5-tetrazine-3-yl)-2,2'-bipyridine (MTB) ligand that has been developed for lanthanide/actinide separation. A multimethod study of the complexation of MTB with trivalent actinide and lanthanide ions is presented. Single-crystal X-ray diffraction measurements reveal the formation of [Ce(MTB)2(NO3)3], [Pr(MTB)(NO3)3H2O], and [Ln(MTB)(NO3)3MeCN] (Ln = Nd, Sm, Eu, Gd). In addition, the complexation of Cm(III) with MTB in solution was studied by time-resolved laser fluorescence spectroscopy. The results show the formation of [Cm(MTB)1-3]3+ complexes, which occur in two different isomers. Quantum chemical calculations reveal an energy difference between these isomers of 12 kJ mol-1, clarifying the initial observations made by time-resolved laser fluorescence spectroscopy (TRLFS). Furthermore, quantum theory of atoms in molecules (QTAIM) analysis of the Cm(III) and Ln(III) complexes was performed, indicating a stronger covalent contribution in the Cm-N interaction compared to the respective Ln-N interaction. These findings align well with extraction data showing a preferred extraction of Am and Cm over lanthanides (e.g., max. SFAm/Eu = 8.3) at nitric acid concentrations <0.1 mol L-1 HNO3.

Solvent effect on the excited state photophysics of Imiquimod: A DFT/TD-DFT and spectroscopic study

Solvation plays an important role in chemistry and biology. The role of the solvent is crucial in any chemical processes such as tautomerization that controls the structure and function of biomolecules. The current study aims to explore how different solvent medium affects the tautomeric forms of an antitumor drug, Imiquimod (IMQ). We have used several spectroscopical methods, including UV–Vis absorption spectroscopy, steady-state and time-resolved fluorescence spectroscopy, and quantum mechanical (QM) calculations. Our results revealed that solvent, indeed, plays a significant role in the modulation of the photophysics of the drug. IMQ has three emission bands in protic and aprotic solvents and two bands in non-polar medium associated with different forms of the drug. Using time-resolved technique, and comparing with the predicted lifetimes from QM calculations, we succeed to assign these three forms as cation, tautomer and neutral of IMQ with 1.6 ns, 2.0 ns and 4.0 ns lifetime values, respectively. Since IMQ is a nucleobase analogue and one of the most effective medications for skin tumors; these findings about which specie of IMQ is present in a given medium, and beyond, how the medium alters the photophysics of the molecule may provide deeper insights into its structure and function.

An inclusive comparison regarding aggregation of surface active ionic liquid and conventional surfactant with a cationic dye Acridine Red exposed in view of spectroscopic and theoretical study

The investigation was conducted on the interaction of acridine red (AR) with sodium dodecyl sulfate (SDS) and 1-butyl-3-methyl imidazolium octyl sulfate (BMImOS) in both premicellar and post micellar concentrations. The interaction between AR and room temperature ionic liquid (RTIL) 1-butyl-3-methyl imidazolium bromide (BMImBr) was studied to understand the effect of imidazolium cation on micellization. Spectroscopic experiments, such as UV–visible absorption spectroscopy, steady-state fluorescence spectroscopy, time-resolved fluorescence spectroscopy, and dynamic light scattering, have been introduced. Various important parameters like spectral shifts, anisotropy, and aggregation number have been determined spectroscopically. However, a fresh insight into density functional theory calculations has also been provided. The observed results have been explained in terms of the aggregation behavior of the amphiphiles. A prominent redshift accompanied by a change in intensity is observed in the presence of surfactant and SAIL. However, in the case of BMImBr, no such spectral shift has been observed; only a decrease in spectral intensity highlights the quenching ability of the imidazolium cation. Average lifetime also provides some additional information regarding the difference in the formation of aggregates by SDS and BMImOS, which is further established from particle size density obtained from DLS. The salt like role of 1-butyl-3-methyl imidazolium cation also has been logically explained by anisotropy measurement and the determination of aggregation number. Using Gauss View 5.0 and Gaussian 09 package, the energy of optimization of each set of amphiphiles along with dye has been determined to follow the interaction at the molecular level. TDDFT calculations have also been performed to determine the structure of HOMO and LUMO.

Steady State and Time-Resolved Fluorescence Spectroscopy of Cinchonine Dication in Sodium Dodecylsulphate Micellar System.

This paper reports the influence of surface charge of the micelles on to the photophysical properties of a cinchonine dication (C2+) fluorophore in anionic, sodium dodecylsulphate (SDS), surfactant at premicellar, micellar and post-micellar concentrations in aqueous phase at room temperature. The magnitude of edge excitation red shift (EERS) in the fluorescence maximum of C2+ in bulk water solution is 1897cm- 1 whereas, in the case of SDS it is observed to be 1984cm- 1. The fluorescence decay curve of C2+ fits with multi exponential functions in the micellar system. The increase in lifetime of C2+ in SDS has been attributed to the increase in radiative rate due to the incorporation of C2+ at the micelle -water interface. The value of dynamic quenching constant determined is 16.9 M- 1. The location of the probe molecule in micellar systems has been justified by a variety of spectral parameters such as dielectric constant, ET (30), viscosity, EERS, average fluorescence decay time, radiative and non-radiative rate constants. All experimental results suggest that the C2+ molecule binds strongly with the SDS micelles and resides at micellar-water interface. The binding constant (Kb) calculated (3.85 × 105 M- 1) for C2+ in SDS revealed that the electrostatic forces mediate charge probe-micelle association.