Methods and Applications in Fluorescence

Efficient and rapid detection of angiotensin-converting enzyme (ACE) activity is important for preventing hypertension and the discovery of new angiotensin-converting enzyme inhibitors (ACEI). In this work, a single-excitation and double-emission biomass-derived carbon quantum dots (CQDs) was prepared and applied for ratiometric fluorescence detection of ACE. Fresh banyan leaves were extracted with ethanol and acetone, and the extracted solution was used as the precursor to produce the carbon quantum dots (BL-CQDs) with single-excitation and double-emission properties...

Fluorescence microscopy can provide valuable information about cell interior dynamics. Particularly, mean squared displacement (MSD) analysis is widely used to characterize proteins and sub-cellular structures' mobility, providing molecular diffusion laws. The MSD curve is traditionally extracted from individual trajectories recorded by single-particle tracking-based techniques. More recently, image correlation methods like iMSD have been shown capable of providing averaged dynamic information directly from images, without the need for isolation and localization of individual particles...

PIFE was first used as an acronym for protein-induced fluorescence enhancement, which refers to the increase in fluorescence observed upon the interaction of a fluorophore, such as a cyanine, with a protein. This fluorescence enhancement is due to changes in the rate of cis/trans photoisomerisation. It is clear now that this mechanism is generally applicable to interactions with any biomolecule and, in this review, we propose that PIFE is thereby renamed according to its fundamental working principle as photoisomerisation-related fluorescence enhancement, keeping the PIFE acronym intact...

Photo-induced dark transient states of fluorophores can pose a problem in fluorescence spectroscopy. However, their typically long lifetimes also make them highly environment sensitive, suggesting fluorophores with prominent dark-state formation yields to be used as microenvironmental sensors in bio-molecular spectroscopy and imaging. In this work, we analyzed the singlet-triplet transitions of fluorescein and three synthesized carboxy-fluorescein derivatives, with one, two or four bromines linked to the anthracence backbone...

In this study, Robinia hispida L. leaves (RH) was used as a precursor for the first time to synthesize fluorescent carbon dots (CDs) with stable blue fluorescence by a single-step hydrothermal synthesis method. Notably, the innovative approach eliminates the necessity for toxic chemicals or hazardous substances, marking a significant advancement in the field. The synthesized CDs demonstrate CDs demonstrates the predominance of spherical shapes with an average size of 11.63 ± 1.92 nm. The CDs not only exhibit an enhanced fluorescent efficiency with a relatively high quantum yield of up to 6...

The challenge of building a highly reliable contactless temperature probe with high sensitivity, good temperature-induced color discriminability, and economical synthesis has prompted the research community to work in the field of rare-earth-based luminescence thermometry. Moreover, the fast-growing market for optoelectronic devices has increased the demand for tunable color-emitting phosphors. In this study, Dy3+/Eu3+ co-doped SrMoO4 phosphors were developed as tunable color-emitting source and dual-mode luminescence thermometer...

The survival rate of oral squamous cell carcinoma (OSCC) patients is very poor, but it can be improved using highly sensitive, specific, and accurate techniques. Autofluorescence and fluorescence techniques are very sensitive and helpful in cancer screening; being directly linked with the molecular levels of human tissue, they can be used as a quantitative tool for cancer detection. Here, we report the development of multi-modal autofluorescence and fluorescence imaging and spectroscopic (MAF-IS) smartphone-based systems for fast and real-time oral cancer screening...

Melamine has been intentionally added into food products to increase the protein count at less cost, especially in dairy products for infant resulting in serious adverse effects on health of consumers. Therefore, this study aimed to develop a method to quantify melamine in dairy products based on the change of fluorescent properties of carbon dots (CDs) as sensing probe. CDs with green-fluorescent emission were synthesized from citric acid and urea under microwave irradiation. The synthesized CDs emitted fluorescence at the maximum wavelength of 538 nm with excitation wavelength of 410 nm...

The stoichiometry of molecular components within supramolecular biological complexes is often an important property to understand their biological functioning, particularly within their native environment. While there are well established methods to determine stoichiometry in vitro, it is presently challenging to precisely quantify this property in vivo, especially with single molecule resolution that is needed for the characterization stoichiometry heterogeneity. Previous work has shown that optical microscopy can provide some information to this end, but it can be challenging to obtain highly precise measurements at higher densities of fluorophores...

Antibodies have gained considerable importance in laboratory and clinical settings. Currently, antibodies are extensively employed for the diagnosis and treatment of several human diseases. Herein, using targeted and cell immunisation approaches, we developed and characterised an antibody clone, DWH24. We found that DWH24 is an IgM κ type antibody that enables excellent visualisation and quantification of dead cells using immunofluorescence, fluorescence microscopy, and flow cytometry. This property was proved by the spontaneous cell death of several tumour cell lines and stimulated T cells, as well as after chemo- and photodynamic therapy...

A green, one-pot, quick, and easily synthesized nitrogen and sulfur co-doped carbon quantum dots (N,S-CDs) were obtained from cheap and readily available chemicals (sucrose, urea, and thiourea) using a microwave-assisted approach in about 4 min and utilized as a turn-off fluorescent sensor for estimation of natamycin (NAT). First, the effect of N and S doping on the microwave-synthesized CDs' quantum yield was carefully studied. CDs derived from sucrose alone failed to produce a high quantum yield; then, to increase the quantum yield, doping with heteroatoms was carried out using either urea or thiourea...

The cell membrane has a fundamental role in the cell life cycle but there's
still much to be learned about its heterogeneous structure, regulation, and protein interaction.
Additionally, the protein-membrane interaction is often overlooked when studying specific protein
dynamics. In this work, we present a new tool for a better understanding of protein dynamics
and membrane function using live cells and fast non-invasive techniques without the need for
individual particle tracking...

A facile, simple, green and sensitive spectrofluorometric method was developed for determination of the calcimimetic drug cinacalcet hydrochloride. It is used for the treatment of hyperparathyroidism. The drug showed high native fluorescence intensity at 320 nm after excitation at 280 nm. The method was linear over the range of 5.0 - 400.0 ng/mL with excellent correlation (R2 = 0.9999). Limit of detection (LOD) and limit of quantitation (LOQ) values were 1.19 and 3.62 ng/mL, respectively. The percentage recovery was found to be 100...

Urine is a highly complex fluorescent system, the fluorescence of which can be affected by many factors, including the often-ignored initial urine concentration in comprehensive fluorescent urine analysis. In this study, a total urine fluorescent metabolome profile (uTFMP) was created as a three-dimensional fluorescence profile of serial synchronous spectra of urine diluted by geometric progression. uTFMP was generated using software designed for this purpose after recalculating the 3D data concerning the initial urine concentration...

Since the small molecular drugs pharmacodynamics and pharmacokinetics could be affected by human serum albumin (HSA) transport, we used spectroscopic and molecular docking studies to investigate the interaction between HSA and the angina-prevention drug trimetazidine (TMZ). As shown by synchronous fluorescence spectroscopy, this interaction affects the microenvironment confirmation around tyrosine residues. The site-competitive experiments showed that TMZ had an affinity toward subdomain III A (site II) of HSA...

The present work investigated the influence of different halides on the excited state dynamics of 6-methoxyflavone (6MF) in an aqueous solution with steady-state and time-resolved techniques. On successive addition of I¯ and Br¯ ions, the fluorescence (FL) of 6MF quenched significantly, whereas the respective ions do not change the maximum fluorescence band. Fluorescence of 6MF was quenched 66 % by I¯ ions and 34 % by Br¯ ions. In a pure aqueous medium, both the H-bonded: CT and protonated species of 6MF participate in the quenching of FL...

Multi-color fluorescence imaging is a powerful tool for studying the spatial relationships and interactions among sub-cellular structures in biological specimens. However, if improperly corrected, geometrical distortions caused by mechanical drift, refractive index mismatch, or chromatic aberration can lead to lower image resolution. In this paper, we present an extension of the image processing framework of Scipion by integrating a protocol called OFM Corrector, which corrects geometrical distortions in real-time using a B-spline-based elastic continuous registration technique...

Enhanced green fluorescence protein (EGFP) is a fluorescent tag commonly used in cellular and biomedical applications. Surprisingly, some interesting photochemical properties of EGFP have remained unexplored. Here we report on two-photon-induced photoconversion of EGFP, which can be permanently converted by intense IR irradiation to a form with a short fluorescence lifetime and spectrally conserved emission. Photoconverted EGFP thus can be distinguished from the unconverted tag by the time-resolved detection...

This work describes the relationship between the complex of photosystem I and photosystem II in the senescence process of rice leaves observed through changes in the optical response. We studied three varieties of rice plants at different aging times using time-resolved photoluminescence to measure the time decay of the emission, and stationary photoluminescence, to measure the emission wavelength. The spectra obtained with the former technique were fitted with decreasing exponential functions. Two relaxation times were obtained, one ranging between 1...

Polyvinylpyrrolidone (PVP) can be used to produce upconversion nanoparticles (UCNPs) in an advantageous manner, i.e. at modest temperatures in open-to-air conditions with simple hotplate and flask apparatus. However, the influence of PVP parameters on the formation of UCNPs has not been previously investigated. In this exploratory study, we establish that PVP molecular weight and relative amount of PVP can greatly influence the morphology and diameter of NaYF4:Yb,Er UCNPs produced via the PVP-assisted route...