Steady state and time-resolved autofluorescence studies of human colonic tissues

Steady state and time-resolved autofluorescence spectroscopies are employed to study the autofluorescence characteristics of human colonic tissues in vitro. The excitation wavelength varies from 260 to 540 nm, and the corresponding fluorescence emission spectra are acquired from 280 to 800 nm. Significant difference in fluorescence intensity of excitation-emission matrices (EEMs) is observed between normal and tumor colonic tissues. Compared with normal colonic tissue, low nicotinamide adenine dinucleotide (phosphate) (NAD(P)H) and flavin adenine dinucleotide (FAD), and high amino acids and protoporphyrin Ⅸ (PpⅨ) fluorescences characterize high-grade malignant tissue. Moreover, the autofluorescence lifetimes of normal and carcinomatous colonic tissues at 635 nm under 397-nm excitation are about 4.32±0.12 and 18.45±0.05 ns, respectively. The high accumulation of endogenous PpⅨ in colonic cancers is demonstrated in both steady state and time-resolved autofluorescence spectroscopies.     

Time-resolved studies of single quantum dots in magnetic fields

We present time-resolved and time-integrated spectroscopy of single InAs quantum dots grown in a GaAs matrix. We observe a number of interesting features in the spectra, including the zero field splitting of exciton and biexciton lines due to quantum dot asymmetry. By the application of an in-plane magnetic field, the normally optically active and inactive exciton states become mixed, enabling us to optically probe the normally inaccessible ‘dark’ states. Time resolved measurements on the mixed states show decay times several times longer than the exciton lifetime at zero field, which we show to be consistent with a dark exciton lifetime orders of magnitude longer than that for bright exciton.     

Emerging biomedical and advanced applications of time-resolved fluorescence spectroscopy

Time-resolved fluorescence spectroscopy is presently regarded as a research tool in biochemistry, biophysics, and chemical physics. Advances in laser technology, the development of long-wavelength probes, and the use of lifetime-based methods, are resulting in the rapid migration of timeresolved fluorescence to the clinical chemistry lab, the patient's bedside, and even to the doctor's office and home health care. Additionally, time-resolved imaging is now a reality in fluorescence microscopy and will provide chemical imaging of a variety of intracellular analytes and/or cellular phenomena. Future horizons of state-of-the-art spectroscopy are also described. Two photon-induced fluorescence provides an increased information content to time-resolved data. Two photoninduced fluorescence, combined with fluorescence microscopy and time-resolved imaging, promises to provide detailed three-dimensional chemical imaging of cells. Additionally, it has recently been demonstrated that the pulses from modern picosecond lasers can be used to quench and/or modify the excited-state population by stimulated emission since the stimulated photons are directed along the quenching beam and are not observed. The phenomenon of light quenching should allow a new class of multipulse time-resolved fluorescence experiments, in which the excited-state population is modified by additional pulses to provide highly oriented systems.     

Excited-state hydrogen bonding effect on dynamic fluorescence of coumarin 102 chromophore in solution: A time-resolved fluorescence and theoretical study

The steady-state absorption and emission as well as the time-resolved fluorescence spectra of coumarin 102 (C102) in both aprotic and alcoholic solvents have been used to study the effect of excited-state hydrogen bond on the dynamic fluorescence of C102 chromophore in various solutions. The dual fluorescence of C102 in alcohols, which is dependent on the hydrogen-bonded donation ability of the solvent, has been assigned to the distribution of free C102 and a hydrogen-bonded complex. Furthermore, a shift of the fluorescence spectra induced by excited-state hydrogen bond has been demonstrated to take place within hundreds of picoseconds by the performance of the time-resolved fluorescence spectra with the time-correlated single-photon-counting (TCSPC) technique. Moreover, the time-dependent density functional theory (TDDFT) has been used to calculate the hydrogen-bonded equilibrium constant pK_HB in different electronic states. It has been demonstrated for the first time that the hydrogen bond strengthening in electronic excited states could decrease the free energy of the hydrogen-bonded complex due to its stronger binding energy. Therefore, the hydrogen-bonded equilibrium will become markedly in favor of the hydrogen-bonded forms in electronic excited states by comparison with the case in the ground state.     

Steady state and time-resolved spectroscopic investigations on the photoreactions involved within the electronically excited electron acceptor 9-cyanoanthracene in presence of benzotriazole and benzimidazole donors

The electrochemical, “steady-state” and “time-resolved” spectroscopic investigations were made on the well-known electron acceptor 9-cyanoanthracene (CNA) when interacted with the electron donors benzotriazole (BZT) and benzimidazole (BMI) molecules. Though electrochemical measurements indicate the thermodynamical possibility of occurrences of photoinduced electron transfer reactions within these reacting systems in the lowest excited singlet state (S₁) of the acceptor CNA but the steady-state and time-resolved measurements clearly demonstrate only the triplet-initiated charge separation reactions. It was reported earlier that in the cases of disubstituted indole molecules the occurrences of photoinduced electron transfer reactions were apparent both in the excited singlet and triplet states of the acceptor 9-cyanoanthracene, but the similarly structured present donor molecules benzotriazole (and benzimidazole) behave differently from indoles. The weak ground state complex formations within the presently studied reacting systems appear to be responsible for the observed static quenching phenomena as evidenced from the time-resolved fluorescence studies. Time-resolved spectroscopic investigations demonstrate the formation of the ground state of the reacting components (donor and acceptor) through recombination of triplet ion-pairs via formations of contact neutral radical produced by H-abstraction mechanism.     

Phonon satellites and time-resolved studies of carrier recombination dynamics in InGaN quantum wells

We have studied the photoluminescence and time-resolved photoluminescence of a set of InGaN quantum wells with well thickness from 1 to 7.5 nm. An analysis of the phonon satellites at 5 K shows Huang–Rhys factors from 0.32 to 0.44. The increase of this factor is caused by the electron–hole separation induced by the piezoelectric field. The time-resolved photoluminescence at room temperature shows that the decay time of the 1 and 2 nm wells does not depend on the wavelength. The maximum decay time is around 600 ps for the 2, 3 and 4 nm wells. However, for the 3 and 4 nm wells a decrease of the photoluminescence decay time is observed at the highest wavelengths. This suggest the onset of a non-radiative process in these samples. The optimum well width for efficient emission for these single quantum wells was found to be 2 nm.     

Characterization of a confocal microscope for time-resolved photon counting fluorescence

A confocal microscope setup is developed for time-resolved fluorescence measurements. It is added to a traditional cuvette time-resolved setup, with a pumped Ti-Sa light source. The temporal resolution of 37 ps (FWHM) is not degraded, in comparison with the cuvette setup also described. These setups allow both decay lifetime and anisotropy relaxation time determination. Fluorescence correlation spectroscopy (FCS) is used to determine the observation point size. When associated with the calcium probe calcium green, calcium concentration in single cells can be determined in 10 ms by simultaneous acquisition of early and late fluorescence photons.     

Dynamics of single InGaN quantum dots

Decay dynamics for single InGaN quantum dots are presented using time-resolved photoluminescence. The recombination is shown to be characterized by a single exponential decay, in contrast to the non-exponential recombination dynamics seen in the 2D wetting layer. The lifetimes of single dots in the temperature range 4–60 K decrease with increasing temperature. Different dots show similar lifetimes of 2 ns.     

X-ray luminescence of Pb-containing microcrystals dispersed in the CsCl Matrix

The process of complexing in CsCl crystals in activating them with lead haloids PbX₂ (X=Cl, Br, I) (C_{PbX 2}=1 mole %) is investigated. The formation of CsPbCl₃ and Cs₄PbCl₆ microcrystals that are dispersed in the CsCl matrix is recorded. The luminescence-kinetic characteristics of the indicated microcrystals and single Pb²⁺v _c ⁻ centers are investigated. It is noted that the formation of the CsPbCl₃ and Cs₄PbCl₆ microcrystals is hindered as the radius of the anion of the lead haloid PbX₂ (X=Cl, Br, I) increases. Translated from Zhurnal Prikladnoi Spektroskopii, Vol. 67, No. 4, pp. 480–482, July–August, 2000.     

Steady state and time-resolved magneto-luminescence in ZnSe/(Zn,Mn)Se Heterostructures

We report on optical measurements performed on two 53Å and 106Å wide ZnSe quantum wells separated by a 350Å thick Zn_0.73Mn_0.27Se barrier. The measurements were performed by means of cw photoluminescence up to 20T, cw photoluminescence excitation and time-resolved photoluminescence spectroscopies up to 9T, at low temperature (4.2K). The fundamental optical transition changes its nature from a type-I light hole excition to a type-II heavy hole exciton as a function of applied magnetic field. Calculations taking into account the strain, Zeeman, and excitonic effects support the experimental findings and allows us to specify the value of the relative valence band offset.     

Steady state and transient fluorescence studies of CaF2: Dy single crystals

The steady state and transient fluorescence spectra of CaF₂: Dy³⁺ have been studied using Ar⁺ and N₂ lasers in the temperature range 77–673 K and in the wavelength region 4500–9000 Å. The spectra show the presence of two types of centers (type A and type B) with decay times of 1.3 and 3.5 ms. The observed spectrum is mostly due to type A centers. Three new groups of fluorescence ⁴F_9/2−⁶H_11/2, ⁴F_9/2−(⁶H_9/2, ⁶F_11/2) and ⁴F_9/2−(⁶H_7/2, ⁶F_9/2) have been observed for type A centers, and a tentative assignment of the positions of the Stark levels of ⁶H_11/2, (⁶H_9/2, ⁶F_11/2) and (⁶H_7/2, ⁶F_9/2) has been made. The change in the intensity of the transitions with temperature and excitation wavelength are discussed.     

Steady-state and time-resolved spectroscopy of porous silicon

We present experimental data on steady-state properties, time-resolved properties and on polarization characteristics of porous silicon photoluminescence and models for the decay processes of the red-orange band. The manifold manifestation of inhomogeneous broadening of this band in emission, excitation, polarization, kinetics and degradation supports the model in which porous silicon is treated as a network of crystallites connected via an oxide interface. Spectral inhomogeneties of the red-orange band can be described in terms of varying shape and size of silicon clusters. The polarization of emission is explained by coexistence of dot-like and wire-like entities, i.e. spherical and non-spherical clusters. The relative weight of these species determines the polarization degree, whereas the kinetics are controlled by the transport of excitations among the clusters. The decay is modeled by a modified stretched exponential function with the local lifetime, the migration lifetime, and a scaling factor. The latter is determined by the dimensionality of the space available for migration which was found to be close to but less than unity. On the nanosecond range two distinct bands in the blue-green region are evaluated that need further studies for interpretation. Generally, arguments are proposed in favor of a quantum confinement origin of the red-orange band and a bridge between quantum-wire and quantum-dot models is provided.     

Prospect for detecting squeezed states of light created by a single atom in free space

We discuss the possibilities of studying in detail the dynamics of spontaneous emission of a single photon by a single atom and measuring the transient degree of squeezing by means of full solid angle fluorescence detection.     

Steady-State and Time-Resolved Fluorescence Spectroscopic Studies on Interaction of the N-terminal Region with the Hairpin Loop of the Phytocystain Scb

The steady-state and time-resolved fluorescece spectroscopy is one of the most powerful method to detect and analyze subtle conformation change and interaction between peptide elements in protein. Phytocystatin Scb isolated from sunflower seeds includes a single Trp residue at position 85. In an attempt to investigate the interaction of the N-terminal region of Scb with the first and second hairpin loops by fluorescence spectroscopy of Trp residue, two Scb mutants in which single Trp locates at position 52 and 58, respectively, and their N-terminal removed mutants were generated. The N-terminal truncation changed the fluorescence decay kinetics of Trp52 from the triple exponential to double. Furthermore, the time-resolved fluorescence anisotropy residue indicated that the segmental motion of Trp52 was significantly enhanced by its N-terminal truncation. In contrast, Trp58 and Trp85 had little influence. The N-terminal successive truncations of Scb and its mutants resulted in the weaken inhibitors to papain. These results suggested that the N-terminal region of Scb interacts with the peptide segment preceding the first hairpin loop, thereby stabilizing the conformation of the hairpin loop structure.     

Effect of annealing on structural and optical properties of lead tungstate microcrystals

Shuttle-like lead tungstate (PbWO₄) microcrystals are synthesized at room temperature using the precipitation method with the cetyltrimethyl ammonium bromide. Results from both the X-ray diffraction and the scanning electron microscopy show that the lattice distortions of the PbWO₄ microcrystals are reduced significantly when the annealing temperature is increased to 873 K. The result from the ultraviolet-visible diffuse reflectance spectroscopy shows that the exciton absorption appears in the sample annealed at 673 K. The self-trapped exciton luminescence due to the Jahn-Teller effect is also observed in the blue band. The interstitial oxygen ions in the WO₄^2- groups are mainly resposible for the enhancement effect of the green luminescence of the annealed samples. The above results are supported by the spectrum analysis of the as-grown and the post-annealed samples using the X-ray photoelectron spectroscopy.     

Steady-State and Time-Resolved Fluorescence Polarization Behavior of Acenaphthene

Steady-state and time-resolved fluorescence polarization studies have been carried out on acenaphthene (ACE) in low-temperature glass solutions and at room temperature. In the low-temperature glass the fluorescence polarization values vary considerably with both emission and excitation wavelength. There is a time dependence (on the nanosecond time scale) of the fluorescence anisotropy, r(t), at 77 K, which has a strong dependence upon the excitation and emission wavelengths. Under these conditions, the time-dependent decay of the anisotropy is not attributable to chromophoric motion. The observations are consistent with emission from two closely lying and interconverting excited states. Rate constants for the photophysical processes involved have been determined by fitting the data using a model proposed by Fleming et. al. The results are discussed with particular reference to the care required in using dynamic fluorescence polarization measurements to determine energy transfer rates in systems containing this chromophore.     

A CTRW-based model of time-resolved fluorescence lifetime imaging in a turbid medium

We develop an analytic model of time-resolved fluorescent imaging of photons migrating through a semi-infinite turbid medium bounded by an infinite plane in the presence of a single stationary point fluorophore embedded in the medium. In contrast to earlier models of fluorescent imaging in which photon motion is assumed to be some form of continuous diffusion process, the present analysis is based on a continuous-time random walk (CTRW) on a simple cubic lattice, the objective being to estimate the position and lifetime of the fluorophore. This can provide information related to local variations in pH and temperature with potential medical significance. Aspects of the theory were tested using time-resolved measurements of the fluorescence from small inclusions inside tissue-like phantoms. The experimental results were found to be in good agreement with theoretical predictions provided that the fluorophore was not located too close to the planar boundary, a common problem in many diffusive systems.     

Sensitized Fluorescence of Silver Nanoparticles in the Presence of Pyrene

The fluorescence of pyrene adsorbed onto the surface of the cetyltrimethylammonium-coated silver nanoparticles was studied. Pyrene molecules adsorbed on freshly prepared silver particles were found to be in close proximity to silver surface thus providing the possibility of energy transfer from excited pyrene to silver cores of the particles. In that case along with the expected fluorescence of pyrene we observed the fluorescence of the silver nanoparticles induced by the excited pyrene molecules. In due course the restructuring of the cetyltrimethylammonium layer resulted in moving of pyrene molecules away from silver surface and simultaneous disappearance of the silver nanoparticles fluorescence band. These data strongly support the recent hypothesis of fluorophore-plasmon coupled emission.     

Steady state fluorescence technique for studying phase transitions in PAAm–PNIPA mixture

The steady state fluorescence (SSF) technique was used to study the sol-gel phase transition during free radical crosslinking copolymerisation of various amounts of acrylamide (AAm) and N-isopropylacrylamide (NIPA) mixtures. N,N′-methylenebis (acrylamide) (BIS) and ammonium persulfate (APS) were used as crosslinker and an initiator, respectively. Pyranine (8-hydroxypyrene-1, 3, 6-trisulfonic acid, trisodium salt, HPTS) was added as a fluoroprobe for monitoring the polymerisation. It was observed that pyranine molecules bind to AAm and NIPA chains upon the initiation of the polymerisation, thus the fluorescence spectra of the bonded pyranines shift to the shorter wavelengths. Fluorescence spectra from the bonded pyranines allowed us to monitor the sol-gel phase transition, without disturbing the system mechanically, and to test the universality of the sol-gel transition as a function of polymer concentration ratios. Observations around the gel point of PAAm–PNIPA mixtures show that the gel fraction exponent β obeyed the percolation result.     

Time-resolved laser-induced saturated fluorescence measurements in a thallium see-through hollow cathode discharge: Evaluation of ground state number density and quantum efficiency

A thallium see-through hollow cathode lamp, or galvatron, was studied to evaluate its potential as a narrow band atomic line filter. Time-resolved laser-induced saturated fluorescence was used to evaluate the ground state number density of this glow discharge as a function of current. It was found to produce a sufficient number density at 16.0 mA to absorb 99.9% of incident light from a line source based on an absorption curve-of-growth calculation. A saturation curve was experimentally obtained and modeled with a time-dependent two-level model, as well as a time-dependent three-level model in the presence of a trap. The three-level model showed excellent agreement with the experimental data when a 10 ns pulse duration was used and when collisional rate constants were set to zero. The quantum efficiency of the system was found to be limited only by the spontaneous transition probabilities. Evaluation of these two parameters has shown that a thallium galvatron is an attractive atom reservoir for the applications as a narrow band atomic line filter.