Table of contents

The fluorescence quantum yield is a measure of the efficiency of photon emission and quantifies the luminescent performance of a given sample. The determination of fluorescence quantum yields, particularly in scattering media, is relevant in the areas of materials science, technology and photonics. It is equally crucial when studying fluorescent bioanalytical probes and biological systems either for medical applications, physiological analyses or the interpretation of optical signals in nature. This type of determination represents a challenge since light scattering introduces an appreciable complexity in the measurements. Hence, the use of experimentally accurate methods and the understanding of their basis and principles is indispensable for obtaining reliable results. In addition, light re-absorption processes are usually very significant in these systems and the experimental quantum yields normally differ from the true quantum yields of the fluorophore. The first purpose of this work is to provide a clear and comprehensive compilation of the various optical methods that can be used for the determination of quantum yields in scattering media. A second purpose is to present the correction models to account for light re-absorption processes, applicable in each case. The advantages and disadvantages of each methodology are comparatively discussed, the difference between experimental and true quantum yield is clarified and it is explained which should be used depending on the case. Several examples previously published in literature are illustrated. The methods presented here are adequate for the study of very diverse samples such as suspensions, solid powders, films, animal tissues and even plant material.

DNA-DNA reactions can be monitored with a label-free fluorogenic reaction. Guanosine-rich, single-stranded DNA oligonucleotides bind to thioflavin-T (ThT) and enhance the fluorescence of the dye. We discovered a novel DNA sequence that produces fluorescence upon binding to ThT. We denote this oligonucleotide ThTSignal. We use ThTSignal as a label-free reporter for the activity of several designed DNA-DNA reactions (DNA circuits). The DNA circuits conditionally produce the ThTSignal oligonucleotide by association or by liberating the ThTSignal oligonucleotide from double-stranded DNA. This strategy offers label-free, cost-effective, fluorogenic detection of the molecular beacon reaction, split reporter reaction, one-step strand displacement reaction, and the entropy-driven amplifier reaction (a catalytic DNA circuit).

Protein misfolding and aggregation into amyloid structures is linked with a number of pathophysiological disorders. In the past decade, significant progresses have been made in the drug discovery strategies against toxic aggregates. Although lack of specificity and high sensitivity for in vitro screening system still seen. Here we demonstrate a new targeting probe based on FF diphenylalanine peptide -protected gold nanoclusters (FF AuNCs). Diphenylalanine peptide has previously been shown to self-assemble into well-ordered fiber like the fibers that are observed in amyloid aggregates. We used of the self-assembly properties along with the ability of FF dipeptide in reduction of gold ions for synthesis of novel Au nanoclusters. We used FF AuNCs for monitoring of effectiveness of anti-amyloid drugs. Fluorescence was considerably diminished when drugs at different concentrations added, due to destruction of the amyloid fibers. Furthermore, the analysis of several components demonstrates significant selectivity against the amyloid disrupting molecules. Prepared FF AuNCs will gain possible strategy for in vitro screening of amyloid disrupting molecules.

The use of organic based fluorophores has been firmly established as a key tool in the biological sciences, with many biological-sensing methods taking advantage of Förster Resonance Energy Transfer (FRET) between different fluorescent organic based dyes following one photon excitation. Nevertheless, the employment of UV-visible absorbing dyes as fluorescent tags and markers typically suffer from several drawbacks including relatively high energy of excitation wavelength, photobleaching and competitive autofluorescence, which often limits their effectiveness and longevity both in vitro and in vivo. As an alternative, lanthanide doped upconverting phosphors (UCP) have emerged as a new class of materials for use in optical imaging and RET sensing; they exhibit high photo- and chemical stability and utilise near infrared excitation. Approaches to sensing a given analyte target employing upconverting phosphors can be achieved by engineering the UCP to operate analogously to fluorescent dyes via Luminescence Resonance Energy Transfer (LRET) and such systems are now becoming central to optically sensing low concentrations of biologically important species and performing distance measurements. Similarly to FRET, the LRET process is distance dependent and requires spectral overlap between the absorption of the acceptor luminophore and the emission of the donor moiety, yet essential measures of the relationship between spectral overlap and the degree of quenching have not yet been established. To address this, we have investigated the Stern-Volmer relationship for a set of six commonly functionalised organic dyes and seven biomolecules that contain key chromophoric co-factors with Gd₂SO₄:Yb:Er (PTIR545) and Gd₂SO₄:Yb:Tm (PTIR475) UCPs under low power nIR excitation, and found that for the organic dyes a linear relationship between spectral overlap and degree of quenching is observed. However, this linear relationship is observed to break down for all the biomolecules investigated.

We demonstrate that single functionalized silver nanowires form a geometric platform suitable for efficient real-time detection of single photoactive proteins. By collecting series of images using wide-field fluorescence microscopy, events of single protein attachment can be distinguished with the signal to noise ratio further improved by fluorescence enhancement due to plasmon excitations in the nanowires. The enhancement is evidenced by strong shortening of the fluorescence decay of single photoactive proteins conjugated to the silver nanowires.

Hospital infections associated with multidrug-resistant (MDR) Pseudomonas aeruginosa are a worldwide public health problem. Efflux systems and biofilm formation are mechanisms related to resistance to carbapenemics. In this study, quantum dots (QDs) were used to evaluate the effect of carbonyl cyanide-3-chlorophenylhydrazone (CCCP), an efflux pump system inhibitor, on biofilm formation and antimicrobial resistance profile of P. aeruginosa strains. For this, QDs were covalently conjugated to meropenem (MPM) and incubated with a P. aeruginosa resistant isolate (P118) or a control sensitive strain (ATCC Pa27853). P118 was also analyzed with conjugates after previous CCCP efflux inhibitor incubation. Fluorescence microscopy images showed that both sensitive and resistant bacteria were efficiently labeled. Nevertheless, P118 isolates presented fluorescent cell agglomerates, suggesting biofilm formation. The addition of the CCCP changed the labeling profile of the resistant isolate, and the absence of agglomerates was observed, indicating no biofilm formation. Genetic assays revealed the presence of MexA and MexE genes encoding channel proteins from efflux pump systems in both resistant and sensitive strains. Disk-diffusion and broth microdilution tests determined drug susceptibility profiles in the presence and absence of CCCP for P118 isolates. We verified that the CCCP efflux system inhibitor may contribute to P. aeruginosa resistant phenotype reduction for some antimicrobials. This study verified the efficiency of QD-MPM conjugates to trigger and study biofilm formation, or its inhibition, before and after CCCP addition. QDs conjugated to antimicrobials can be used as nanotools to investigate multidrug-resistant bacterial strains on biofilm formation.

Multivariate Curve Resolution with Alternating Least Squares (MCR-ALS) was for the first time successfully used to evaluate an intricate photophysical behavior, where deprotonation on the electronic ground state (S₀), intra and intermolecular proton transfer processes (ESPT and ESIPT) on the electronic excited state (S₁) can simultaneously be presented. In this sense, the organic dye 2-(2'-hydroxyphenyl)benzothiazole (HBT) was used as a proof-of-concept model, where MCR-ALS showed to be a powerful tool for discriminate chemical reactions that occur concomitantly on different potential energy surfaces, which include photochemical reactions. As a result, the chemometric method showed to be a straightforward approach for the determination of the acidic strengths of those equilibria were estimated as 8.61 and 1.11 to hydroxyl deprotonation on electronic ground and excited states, respectively.

There is an increasing need for the development of probes for the detection of hexavalent chromium since it is a known carcinogen, which can cause adverse effects on human health. Metal-organic frameworks (MOFs) have shown successful detection and removal of hazardous substances from aqueous media. This work presents the use of simple organic ligands such as 3-pyridinecarboxaldehyde and trimesic acid with Zn(II) ion to fabricate a new MOF that exhibits sensitive and selective luminescence quenching response towards CrO₄²⁻ and Cr₂O₇²⁻ species in aqueous solution. The MOF showed a detection limit of 0.67 μM (0.078 ppm) as CrO₄²⁻ species and 1.91 μM (0.41 ppm) as Cr₂O₇²⁻ species. Results reveal that the as-synthesized MOF could serve as a good luminescent sensor for CrO₄²⁻ and Cr₂O₇²⁻ species in the contaminated aqueous phase.

CsPbBr₃ colloidal quantum dots have been synthesized by hot-injection method showing spherical shape with an average diameter of ∼10.5 nm. UV–vis absorption of CsPbBr₃ colloidal quantum dots shows a broad spectrum with an optical bandgap of ∼2.3682 eV. The steady-state photoluminescence measurement reveals a narrow emission peak at 2.352 eV with full-width at half maximum of 0.113 eV. Absolute photoluminescence quantum yield of colloidal quantum dots dispersed in poly(methyl methacrylate) was found to be 60 ± 1%. The time-resolved photoluminescence data recorded at 266 nm excitation were well fitted using a mono-exponential curve with a decay time of 25.36 (5) ns. A luminescent solar concentrator was fabricated using colloidal quantum dots in transparent poly(methyl methacrylate) polymer uniformly coated over glass substrate that shows an external optical conversion efficiency of ∼5.4% under one sun illumination. The experimental results presented in this manuscript reveals that luminescent solar concentrator prepared using colloidal CsPbBr₃ quantum dots shows absorption in wide spectral range, high absorption coefficient, high photoluminescence quantum yield, high external optical conversion efficiency, and good photostability, thermal stability and long-term stability under ambient conditions and therefore are in many ways superior to the other luminescent materials explored for LSC devices.

The accurate, periodically updated excitation beam intensity correction is essential for conducting fluorescence spectroscopy research. This article describes a simple and inexpensive approach to reevaluate the excitation calibration curve of a spectrofluorometer using a single dye solution. The method shows excellent agreement with the data obtained using a certified calibration detector for the broad spectral range from 290 nm to 700 nm.