Fluorescence Decay Time Research Articles

Fluorescence lifetime-based sensing of sodium by an optode.

We report a 1,2,3-triazol fluoroionophore for detecting Na(+) that shows in vitro enhancement in the Na(+)-induced fluorescence intensity and decay time. The Na(+)-selective molecule 1 was incorporated into a hydrogel as a part of a fiber optical sensor. This sensor allows the direct determination of Na(+) in the range of 1-10 mM by measuring reversible fluorescence decay time changes.

Pump Power Depreciation by Photodarkening in Ytterbium-Doped Fibers and Amplifiers

In this letter, pump power depreciation due to photodarkening effects in an ytterbium doped fiber (YDF) is investigated. Photodarkening is monitored by measuring the transmitted power at visible and pump wavelengths simultaneously, which renders direct quantitative measurement of pump power reduction rather than excess background loss. Our observation of substantial reduction in the pump power and the shortened fluorescence decay time in the photodarkened fiber indicates that the photodarkening influences pump efficiency. The effect of the photodarkening in a device level is also investigated by observing the performance of the YDF amplifier implementing the same YDF. The pump power depreciation of this fiber amplifier is quantitatively investigated by numerical simulation. The results suggest that operating at a saturated regime helps to subdue the photodarkening, with explanation in terms of population inversion density provided. The level of power depreciation is calculated to be 15%-30%, which matches our experimental observation.

PsbS is required for systemic acquired acclimation and post-excess-light-stress optimization of chlorophyll fluorescence decay times in Arabidopsis

PsbS is required for systemic acquired acclimation and post-excess-light-stress optimization of chlorophyll fluorescence decay times in Arabidopsis

Cavity-sensitive amplified spontaneous emission with radiation reabsorption

We experimentally and numerically investigate amplified spontaneous emission (ASE) in the presence of radiation reabsorption by using an end-pumped ruby laser. From the fluorescence time decay in the axial direction, we find that the average path length of ASE is much larger than the reabsorption length and exhibits sharp variation near the semi-concentric cavity configuration; however, the side lifetime is independent of cavity configuration. In addition, an axial power bump corresponding to a side power valley and the enhanced fluorescence R-line spectrum are observed and explained.

The role of the surrounding polarity on the phototautomerization process in a diazaaromatic compound: An UV–vis and NMR study

Fluorescence steady-state and decay measurements of the azaaromatic derivative norharmane were obtained in solvents with different polarities: dichloromethane (DCM) and acetonitrile (ACN), at different temperatures, in the absence and the presence of a donor/acceptor hydrogen bond such as acetic acid. 2D NOESY experiments confirmed that this compound forms a hydrogen-bonding cyclic complex with acetic acid in the ground state. This complex induced a phototautomeric equilibrium, which produced new emission bands at 400 and 500nm. This tautomeric process was established only in the first excited state via a concerted proton–electron transfer. Indeed, the cyclic complex formed in the ground state evolved to a like-cation upon cooling from room temperature to 225K, and a new emission band at 450nm is observed. The fluorescence decay times confirmed these results.

Optical Biopsy of Benign and Malignant Tissue by Time Resolved Spectroscopy

Pathological condition of malignant tissue could be analyzed by spectral domain or time domain spectroscopy, the two being the complementary to each other in optical biopsy (OB) of cancer. This paper reports results of time resolved emission spectroscopy (TRS) of 24 excised tissue samples of breast and prostate (normal control = 12; benign = 4; malignant = 8), employing a 390 nm, 100 fs, Ti-Sapphire laser pulses.The fluorescence decay times were measured using streak camera and the resultant data were fitted for single and bi-exponential decays with reliability of 97%. Our results show the distinct difference between normal, benign and malignant tissues mostly due to the emission spectra of Nicotinamide Adenine Dinucleotide (NADH), Flavin Mononucleotide (FAD) and also due to the heterogeneity of micro environments associated with the diseased tissues. In this short report, fit is also shown that TRS of breast tissues are similar to those of prostate tissues.

Growth, crystal structure and optical study of Tm:LuYSiO5 single crystal

The crystal growth, structure, absorption spectra, J–O parameters, emission spectra, Gain spectra and fluorescence lifetime of Tm:LuYSiO5 single crystal were discussed for the first time to my best knowledge. The absorption cross-section is 0.91×10−20cm2. The full width half maximum is 8nm at 791nm. The larger emission cross-section of 0.27×10−20cm2 at 1832nm and broader full width half maximum of 277nm make Tm:LuYSiO5 suitable for laser output around the 2.0μm wavelength. The fluorescence decay time of the 3F4–3H6 transition was measured to be 1.44ms. All the results indicate that Tm:LuYSiO5 should have potential technological applications in 2μm tunable laser.

Investigation of ground and excited state photophysical properties of gadolinium phthalocyanine

In this present work, we investigated the singlet excited state absorption (ESA) spectrum and intersystem crossing dynamics of gadolinium phthalocyanine (GdPc) in toluene. For that, we employed the femtosecond wavelength-tunable Z-scan and picosecond Pulse Train Fluorescence (PTF) techniques to determine, respectively, the singlet ESA cross-section on a wide spectral range (from 460 up to 740 nm) and the intersystem crossing time (singlet–triplet transition). We observed that the ESA spectrum presents distinct absorption behaviors, i.e., saturable (SA) and reverse saturable absorption (RSA). We have also been able to identify regions in which the excited state absorption cross-section has the same magnitude as the ground state one (at 600 and 720 nm). The RSA effect was observed for wavelengths shorter than 600 nm and longer than 720 nm, while SA occurs in the Q-band region, located between 600 and 720 nm. To describe the main singlet–singlet transitions, we measured the fluorescence decay time and employed a rate equation model considering a three-energy-level-diagram. Ratios between excited and ground state absorption cross-section from 0.4 to 4 were observed along the ESA spectrum. From these results, we calculated the transition dipole moment between the excited states for the main electronic transitions of GdPc. To obtain the intersystem crossing time, we incorporated an additional level in our model and used the PTF technique to probe the population transfer to the triplet state. Our results pointed out that the intersystem crossing time is higher than 300 ns, which corresponds to a maximum triplet quantum yield of 1.5%.

Remote optical sensor for real-time residual salt monitoring on road surfaces

We report on an optical sensor for remote residual dry salt (NaCl) monitoring, with applications in road winter maintenance. The novelty of the sensing principle is the simultaneous measurement of the amplitude and decay time of the natural fluorescence of salt using synchronous detection. We present the characterisation of the luminous properties of salt that allow a sensor design with high sensitivity and stable measurements that match the requirements for the intended applications (concentrations below 20g/m2). The proposed sensor system has a compact design and short response time, capable of being mounted on-board maintenance vehicles for real-time road condition assessment.

Scintillation properties of quantum-dot doped styrene based plastic scintillators

We fabricated quantum-dot doped plastic scintillators in order to control the emission wavelength. We studied the characterization of the quantum-dots (CdSe/ZnS) and PPO (2, 5-diphenyloxazole) doped styrene based plastic scintillators. PPO is usually used as a dopant to enhance the scintillation properties of organic scintillators with a maximum emission wavelength of 380nm. In order to study the scintillation properties of the quantum-dots doped plastic scintillators, the samples were irradiated with X-ray, photon, and 45MeV proton beams. We observed that only PPO doped plastic scintillators shows a luminescence peak around 380nm. However, both the quantum-dots and PPO doped plastic scintillators shows luminescence peaks around 380nm and 520nm. Addition of quantum-dots had shifted the luminescence spectrum from 380nm (PPO) toward the region of 520nm (Quantum-dots). Emissions with wavelength controllable plastic scintillators can be matched to various kinds of photosensors such as photomultiplier tubes, photo-diodes, avalanche photo-diodes, and CCDs, etc. Also quantum-dots doped plastic scintillator, which is irradiated 45MeV proton beams, shows that the light yield of quantum-dots doped plastic scintillator is increases as quantum-dots doping concentration increases at 520nm. And also the plastic scintillators were irradiated with Cs-137 γ-ray for measuring fluorescence decay time.

Time-resolved fluorescence microscopy for quantitative Ca2+ imaging in living cells

Calcium (Ca(2+)) is a ubiquitous intracellular second messenger and involved in a plethora of cellular processes. Thus, quantification of the intracellular Ca(2+) concentration ([Ca(2+)]i) and of its dynamics is required for a comprehensive understanding of physiological processes and potential dysfunctions. A powerful approach for studying [Ca(2+)]i is the use of fluorescent Ca(2+) indicators. In addition to the fluorescence intensity as a common recording parameter, the fluorescence lifetime imaging microscopy (FLIM) technique provides access to the fluorescence decay time of the indicator dye. The nanosecond lifetime is mostly independent of variations in dye concentration, allowing more reliable quantification of ion concentrations in biological preparations. In this study, the feasibility of the fluorescent Ca(2+) indicator Oregon Green Bapta-1 (OGB-1) for two-photon fluorescence lifetime imaging microscopy (2P-FLIM) was evaluated. In aqueous solution, OGB-1 displayed a Ca(2+)-dependent biexponential fluorescence decay behaviour, indicating the presence of a Ca(2+)-free and Ca(2+)-bound dye form. After sufficient dye loading into living cells, an in situ calibration procedure has also unravelled the Ca(2+)-free and Ca(2+)-bound dye forms from a global biexponential fluorescence decay analysis, although the dye's Ca(2+) sensitivity is reduced. Nevertheless, quantitative [Ca(2+)]i recordings and its stimulus-induced changes in salivary gland cells could be performed successfully. These results suggest that OGB-1 is suitable for 2P-FLIM measurements, which can gain access to cellular physiology.

Corneal Metabolic State Assessment by Fluorescence Lifetime Imaging Microscopy

A long time objective of ophthalmologists is to diagnose corneal cells dysfunction prior to its pathologic expression. With this motivation, we are currently developing a new instrument for in vivo metabolic imaging of corneal tissues.Metabolic alterations are known to be the first sign of several corneal pathologies and can be assessed through non-invasive monitoring of metabolic co-factors flavin adenine dinucleotide (FAD) and nicotinamide adenine dinucleotide (NADH). The quantification of the relative proportions between free and protein-bound NADH and FAD can be achieved using fluorescence lifetime-resolved methods. This approach has already been applied in age-related macular degeneration, diabetic retinopathy and epithelial cancer.FAD and NADH imaging can be performed by one-photon excitation (1PE) and two-photon excitation (2PE) techniques. The latest has the advantage of allowing simultaneous excitation of both metabolic co-factors. However, there are still safety concerns when considering in vivo ocular studies in humans using 2PE.Due to these concerns we used, as a first approach, a 1PE system for evaluating the feasibility of corneal FAD imaging. The use of FAD has advantages over NADH. It can be excited over longer excitation wavelengths, is more resistant to photo-bleaching and is located exclusively in the mitochondrial space.A PicoQuant MicroTime 100 (PicoQuant GmbH, Berlin, Germany) coupled to an Olympus BX51 Microscope (Olympus Corporation, Tokyo, Japan) was used to monitor FAD autofluorescence. The instrument uses a 440 nm pulsed diode laser (330 ps) running at a pulse rate of 40 MHz. The instrument was modified by us to allow the acquisition of both fluorescence lifetime and reflectance images and to enhance scattered light rejection.Intensity decay curves were processed with SymPhoTime v5.3 Software (PicoQuant GmbH, Berlin, Germany). The fluorescence decay times were obtained after applying a non-linear least square fit to the decay data and the goodness of fit was evaluated by the analysis of the residuals and the chi-squared (χ2).We have acquired fluorescence lifetime images of ex vivo healthy Wistar rat corneas (Fig.1) using two different instrument setups: 1- using the emission filters provided by the manufacturer; 2- placing extra emission filters to fully reject the scattered excitation light. In both setups, FAD fluorescence data presented a bi-exponential decay with a short (protein-bound FAD) and a longer (free FAD) lifetime component.While both setups provide FAD fluorescence decays, only the second retrieves valid metabolic information. We obtained two lifetime components, one of 0.118 (0.028) ns and another of 2.11 (0.16) ns, with a relative contributions of 39.4 (2.2) and 60.6 (2.2), respectively. These values are in accordance with the literature.Corneal layer discrimination is possible based on morphologic characteristics. However, the fluorescence lifetime images do not provide morphological detail (Fig.1), possibly because FAD is only present in the mitochondria. These organelles are small and tend to accumulate around the nuclei.So, we modified the instrument’s optical setup to allow the acquisition of both fluorescence lifetime images and reflectance images. Figure 2 shows an example of the corneal epithelial layer.The image resolution and depth penetration are still not ideal. Since the assessment of corneal endothelial layer metabolic function is also within our goals, we are currently implementing further modifications to improve both the instrument’s resolution and depth penetration.The characterization of FAD fluorescence lifetime in unhealthy corneas is important to detect corneal dysfunctions prior to its pathologic expression. Therefore, we intend to study metabolic altered Wistar rat corneas. The alterations will be induced by potassium cyanide, which is a reversible inhibitor of the fourth complex of the mitochondrial electron transport chain.Financial support received from the Fundação para a Ciência e a Tecnologia under the research projects PTDC/SAU-BEB/104183/2008 and PTDC/SAU-ENB/122128/2010.

Effects Of Crystallite Grain Size On The Triboluminescent Emmision For EuD4TEA

One of the brightest tested triboluminescent materials is europium dibenzoylmethide triethylammonium (EuD4TEA). In 2011, the authors discovered that synthesizing EuD4TEA using europium nitrate instead of chloride significantly increased the triboluminescence yield. In 2012, the authors discovered that anhydrous ethanol is not the best solvent to be used for synthesizing EuD4TEA. The less expensive acetone increases the triboluminescence and makes the synthesis much easier. However, no study has ever been done on how increasing the crystal size of EuD4TEA changes its triboluminescence. This paper reports the effect of increasing crystal size on the triboluminescence of EuD4TEA. The results indicate that acetone can be used to successfully control the grain size of EuD4TEA and thus increase the amount of triboluminescence emitted. The normalized TL yield is seen to be directly proportional to the square of the average grain size. The fluorescence decay time remains unchanged with an average value of 534.5 ± 7.3 µs, with no change in the emission properties of EuD4TEA. Copyright © 2013 VBRI press.

Multiphoton laser tomography and fluorescence lifetime imaging of melanoma: morphologic features and quantitative data for sensitive and specific non-invasive diagnostics.

Multiphoton laser tomography (MPT) combined with fluorescence lifetime imaging (FLIM) is a non-invasive imaging technique, based on the study of fluorescence decay times of naturally occurring fluorescent molecules, enabling a non-invasive investigation of the skin with subcellular resolution. The aim of this retrospective observational ex vivo study, was to characterize melanoma both from a morphologic and a quantitative point of view, attaining an improvement in the diagnostic accuracy with respect to dermoscopy. In the training phase, thirty parameters, comprising both cytological descriptors and architectural aspects, were identified. The training set included 6 melanomas with a mean Breslow thickness±S.D. of 0.89±0.48 mm. In the test phase, these parameters were blindly evaluated on a test data set consisting of 25 melanomas, 50 nevi and 50 basal cell carcinomas. Melanomas in the test phase comprised 8 in situ lesions and had a mean thickness±S.D. of 0.77±1.2 mm. Moreover, quantitative FLIM data were calculated for special areas of interest. Melanoma was characterized by the presence of atypical short lifetime cells and architectural disorder, in contrast to nevi presenting typical cells and a regular histoarchitecture. Sensitivity and specificity values for melanoma diagnosis were 100% and 98%, respectively, whereas dermoscopy achieved the same sensitivity, but a lower specificity (82%). Mean fluorescence lifetime values of melanocytic cells did not vary between melanomas and nevi, but significantly differed from those referring to basal cell carcinoma enabling a differential diagnosis based on quantitative data. Data from prospective preoperative trials are needed to confirm if MPT/FLIM could increase diagnostic specificity and thus reduce unnecessary surgical excisions.

Fluorescent probe for visualizing guanine-quadruplex DNA by fluorescence lifetime imaging microscopy

The importance of guanine-quadruplex (G4) is not only in protecting the ends of chromosomes for human telomeres but also in regulating gene expression for several gene promoters. However, the existence of G4 structures in living cells is still in debate. A fluorescent probe, 3,6-bis(1-methyl-2-vinylpyridinium) carbazole diiodide (o-BMVC), for differentiating G4 structures from duplexes is characterized. o-BMVC has a large contrast in fluorescence decay time, binding affinity, and fluorescent intensity between G4 structures and duplexes, which makes it a good candidate for probing G4 DNA structures. The fluorescence decay time of o-BMVC upon interaction with G4 structures of telomeric G-rich sequences is ∼2.8 ns and that of interaction with the duplex structure of a calf thymus is ∼1.2 ns. By analyzing its fluorescence decay time and histogram, we were able to detect one G4 out of 1000 duplexes in vitro. Furthermore, by using fluorescence lifetime imaging microscopy, we demonstrated an innovative methodology for visualizing the localization of G4 structures as well as mapping the localization of different G4 structures in living cells.

ChemInform Abstract: Growth and Luminescence Properties of Undoped Strontium Iodide Crystals.

AbstractHigh‐quality SrI2 crystals are grown by the vertical Bridgman method and optical transmission, photoluminescence, radioluminescence, fluorescence, and scintillation decay time data are determined.

Spectral properties and energy transfer in Er3+/Yb3+ co-doped LiYF4 crystal

Laser crystals of LiYF4 (LYF) singly doped with Er3+ in 2.0% and co-doped with Er3+/Yb3+ in about 2.0%/1.0% molar fraction in the raw composition are grown by a vertical Bridgman method. X-ray diffraction (XRD), absorption spectra, fluorescence spectra and decay curves are measured to investigate the structural and luminescent properties of the crystals. Compared with the Er3+ singly doped sample, obviously enhanced emission at 1.5 μm wavelength and green and red up-conversion emissions from Er3+/Yb3+ co-doped crystal are observed under the excitation of 980 nm laser diode. Meanwhile, the emission at 2.7 μm wavelength from Er3+ singly doped crystal is reduced. The fluorescence decay time ranging from 18.60 ms for Er3+ singly doped crystal to 23.01 ms for Er3+/Yb3+ co-doped crystal depends on the ionic concentration. The luminescent mechanisms for the Er3+/Yb3+ co-doped crystals are analyzed, and the possible energy transfer processes from Yb3+ to Er3+ are proposed.

Real-Time Phosphate Sensing in Living Cells using Fluorescence Lifetime Imaging Microscopy (FLIM)

Phosphate ions play important roles in signal transduction and energy storage in biological systems. However, robust chemical sensors capable of real-time quantification of phosphate anions in live cells have not been developed. The fluorescein derivative dye 9-[1-(2-methyl-4-methoxyphenyl)]-6-hydroxy-3H-xanthen-3-one (2-Me-4-OMe TG) exhibits the characteristic excited-state proton-transfer (ESPT) reaction of xanthenic derivatives at approximately physiological pH resulting in the dependence of the dye's nanosecond fluorescence decay time on the phosphate buffer concentration. This allows the 2-Me-4-OMe TG dye to be used with fluorescence lifetime imaging microscopy (FLIM) as a real-time phosphate intracellular sensor in cultured cells. This methodology has allowed the time course of cellular differentiation of MC3T3-E1 murine preosteoblast cells to be measured on the basis of the decrease in the decay time of 2-Me-4-OMe TG. These changes were consistent with increased alkaline phosphatase activity in the extracellular medium as a marker of the differentiation process.

Optical imaging for brain tissue characterization using relative fluorescence lifetime imaging

An autofluorescence lifetime wide-field imaging system that can generate contrast in underlying tissue structures of normal and malignant brain tissue samples with video rate acquisition and processing time is presented. Images of the investigated tissues were acquired with high resolution (∼35 μm) using an algorithm to produce contrast based on differences in relative lifetimes. Sufficient contrast for delineation was produced without the computation of fluorescence decay times or Laguerre coefficients. The imaged tissues were sent for histological analysis that confirmed the detected imaged tissues morphological findings and correlations between relative lifetime maps and histology identified.

Time resolved spectroscopy and gain studies of Fullerenes C60 and C70

The fluorescence decay time of Fullerenes C60 and C70 in pure form as well as in mixture with Coumarin C440 and Quinizarine dyes are studied. Results indicate that the decay of pure fullerenes is constant throughout the solute concentration and it is also independent of excitation wavelength, whereas in the case of mixture with dyes different behavior is noticed. We have also calculated the Stern–Volmer quenching constant and optical gain of both the fullerenes from which it is found that the optical gain is positive for Fullerene C70 only in a very narrow range of concentration.