Luminescent lanthanide chelate contrast agents and detection of lesions in the hamster oral cancer model

Lanthanide chelates are a somewhat unique class of molecules that have proven to be useful in the biomedical field due to their extremely large Stokes' shift and long fluorescent lifetimes. The ability of these molecules to produce fluorescence in the low- or zero-back-ground regime makes this class of molecules excellent candidates for use as contrast agents for a wide variety of applications in biological settings. Here we present the preparation, spectroscopic characterization, and application of a new terbium chelate contrast agent, based on the 1,4,7,10-tetraazacyclododecane macrocycle (cyclen), for detection of early-stage malignant lesions in the Syrian hamster cheek pouch. Tb-P(CTMB) delivers bright blue-green luminescence when excited with low photon fluxes of UV light. As a pilot study, the DMBA-treated Golden Hamster Cheek pouch epithelial cancer model was employed and Tb-P(CTMB) was used as a topical agent for the visual detection of diseased tissue. In this preliminary study the agent tended to associate with early-stage malignant lesions, suggesting that Tb-P(CTMB) could be used as a contrast agent to aid in identifying early-stage oral cancer lesions.     

Studying biological tissue with fluorescence lifetime imaging: microscopy,endoscopy, and complex decay profiles

We have applied fluorescence lifetime imaging (FLIM) to the autofluorescence of different kinds of biological tissue in vitro, including animal tissue sections and knee joints as well as human teeth, obtaining two-dimensional maps with functional contrast. We find that fluorescence decay profiles of biological tissue are well described by the stretched exponential function (StrEF), which can represent the complex nature of tissue. The StrEF yields a continuous distribution of fluorescence lifetimes, which can be extracted with an inverse Laplace transformation, and additional information is provided by the width of the distribution. Our experimental results from FLIM microscopy in combination with the StrEF analysis indicate that this technique is ready for clinical deployment, including portability that is through the use of a compact picosecond diode laser as the excitation source. The results obtained with our FLIM endoscope successfully demonstrated the viability of this modality, though they need further optimization. We expect a custom-designed endoscope with optimized illumination and detection efficiencies to provide significantly improved performance.     

Preparatory study for detection of nickel in industrial flue gas by excimer laser-induced fragmentation fluorescence spectroscopy

The purpose of this work is to survey possibilities for detecting molecular nickel species in industrial flue gas using excimer laser-induced fragmentation fluorescence (ELIF), in particular to establish suitable detection schemes and to obtain a sensitivity estimate for Ni detection. Investigations were conducted in a heated laboratory cell under defined conditions of temperature and pressure, using NiCl2 as the precursor molecule. An ArF excimer laser (193 nm) was used for excitation and Ni atomic emission spectra were recorded in the range 300 to 550 nm. The dependence of ELIF signal on laser fluence was quadratic in the range of laser intensities investigated, as expected for a two-photon excitation process. The temporal behavior of the ELIF signals gave lifetimes significantly longer than the known natural lifetimes. This result and the energetics of the system suggest a Ni* production mechanism involving the formation of Ni+ and subsequent ion-electron recombination. The temperature dependence of the ELIF signal, determined in the range 773 to 1223 K, was found to follow the vapor-pressure curve (Antoine equation) known from the literature. Finally, quenching effects were investigated by measuring ELIF signals and lifetimes in nitrogen or air up to 1 atm. On the basis of the results so far, detection limits for Ni in practical combustion applications in the range of tens of ppb should be achievable, which will be sufficient for regulatory measurements in incinerators and power plants.     

Error analysis of the rapid lifetime determination method for double-exponential decays and new windowing schemes

The rapid lifetime determination method (RLD) is a mathematical technique for extremely rapid evaluations of lifetimes in exponential decays. It has been applied in luminescence microscopy and single-molecule lifetime evaluation. To date, the primary application has been in single-exponential evaluations. We present extensions of the method to double exponentials. Using Monte Carlo simulations, we assess the performance of both the double-exponential decay with known lifetimes and the double-exponential decay with unknown preexponential factors and lifetimes. Precision is evaluated as a function of the noise level (Poisson statistics), the ratios of the lifetimes, the ratios of their preexponential factors, and the fitting window. Optimum measurement conditions are determined. RLD is shown to work well over a wide range of practical experimental conditions. If the lifetimes are known, the preexponential factors can be determined with good precision even at low total counts (10(4)). With unknown preexponential factors and lifetimes, precisions decrease but are still acceptable. A new gating scheme (overlapped gating) is shown to offer improved precision for the case of a single-exponential decay. Theoretical predictions are tested against actual experimental data from a laser-based lifetime instrument.     

Lifetime measurements for random loading in the very high cycle fatigue range

The ultrasound technique (20 kHz loading freqeuncy) has been investigated as a time- and energy-saving method for measuring lifetimes under service loading conditions. AISI C1020 steel was tested between 3 × 10⁶ and 3 × 10⁹ cycles with two Gaussian-like random loading programs (Gauss distribution generated with a Markov matrix and straight-line distribution). The resulting lifetime curves can be approximated by straight lines in a log-log plot. If the maximum values are plotted, these lines lie above the S/N curves and have approximately the same slope as the S/N curve for finite lifetimes. The experimentally found lifetimes are compared with predictions according to Miner and the Miner-Haibach rule (with half the slope of the S/N curve in the endurance range). Good agreement is found for measured and calculated results according to the Miner-Haibach rule if the measured amplitude distributiom is introduced into the calculations. This agreement is especially good for Markov random loading. Predictions according to the original Miner's rule give lifetimes that are too long in the very high cycle range. This result is explained by damaging effects of amplitudes below the endurance limit. For linear distribution random loading this effect has not been observed in the cycle range up to 3 × 10⁹.     

Direct detection of hydrocarbon displacement in a model porous soil with magnetic resonance imaging

The direct detection of hydrocarbon fluid and the discrimination of water through carbon-13 magnetic resonance imaging (MRI) would be a significant advance in many scientific fields including food, petrogeological, and environmental sciences. Carbon-13 MRI is a noninvasive analytical technique that has great potential for direct detection of hydrocarbons. However, the low natural abundance of carbon-13, low gyromagnetic ratio, and generically short transverse signal lifetimes in realistic porous media all conspire to hinder carbon-13 MRI. A multiple echo pure phase encode MRI technique introduced in this paper helps to overcome these limitations. As a pure phase encode technique, it is immune to artifacts arising from inhomogeneous B0 fields. It is also, by its nature, more quantitative than most MRI methods. Viscous hydrocarbon flow through a sand bed, a simple realistic porous medium, was used as our test system. Flow in this model system was driven by capillary suction. The detection limit, spatially resolved, was determined to be 26 mg.     

Hypersensitive luminescence of Eu3+ in dimethyl sulfoxide as a new probing for water measurement

Dimethyl sulfoxide (DMSO) is a well-known organic solvent that can be used for biological applications. DMSO is miscible with water, and it is very common that the two solvents are mixed for some applications. It is important to detect water in DMSO, and this has been done using the luminescence decay lifetimes from Eu(3+) ions. We observed that the emissions of Eu(3+) in DMSO are very strong and very sensitive to water. The emission band from the (5)D(0) → (7)F(2) transition has two peaks at 613 and 617 nm, respectively, and these two peaks change in the opposite ways when water is added into DMSO. The intensity ratio of the two peaks follows nearly perfect linear dependence on the water concentration added in DMSO. This linear relationship provides a new and convenient method for water measurement in DMSO.     

Direct detection of adenosine in undiluted serum using a luminescent aptamer sensor attached to a terbium complex

Aptamers, single-stranded nucleic acids that can selectively bind to various target molecules, have been widely used for constructing biosensors. A major challenge in this field, however, is direct sensing of analytes in complex biological media such as undiluted serum. While progress has been made in developing an inhomogeneous assay by using a preseparation step to wash away the interferences within serum, a facile strategy for direct detection of targets in homogeneous unprocessed serum is highly desired. We herein report a turn-on luminescent aptamer biosensor for the direct detection of adenosine in undiluted and unprocessed serum, by taking advantage of a terbium chelate complex with long luminescence lifetime to achieve time-resolved detection. The sensor exhibits a detection limit of 60 μM adenosine while marinating excellent selectivity that is comparable to those in buffer. The approach demonstrated here can be applied for direct detection and quantification of a broad range of analytes in biological media by using other aptamers.     

Biological sensor based on a lateral electric field-excited resonator

This paper describes a biological sensor based on a lateral electric field-excited resonator using an X-cut lithium niobate plate. Its potential was shown through the example of biological interaction between bacterial cells and specific bacteriophages. The detection was based on the analysis of the measured real and imaginary parts of electrical impedance for a resonator loaded by the biological suspension under study. It has been shown that the sensor is sensitive to specific interactions between bacterial cells and specific bacteriophages in a pure state as well as in the presence of extraneous microflora. The degree of electrical impedance variation resulting from the biological interaction depends on the numbers of phage particles and bacteria cells. The sensor may be used not only for the qualitative analysis of bacteria but also for their quantitative detection.     

High-resolution near-infrared imaging of DNA microarrays with time-resolved acquisition of fluorescence lifetimes

Ultrasensitive, near-infrared (NIR), time-resolved fluorescence is evaluated as a detection method for reading DNA hybridization events on solid surfaces for microarray applications. In addition, the potential of mulitiplexed analyses using time-resolved identification protocols is described. To carry out this work, a NIR time-resolved confocal imager was constructed to read fluorescence signatures from the arrays. The device utilized a 780-nm pulsed diode laser, a single-photon avalanche diode (SPAD), and a high-numerical-aperture microscope objective mounted in an epi-illumination format. Due to the small size of the components that are required to construct this imager, the entire detector could easily be mounted on high-resolution translational stages and scanned over the stationary arrays. The instrument response function of the device was determined to be 275 ps (fwhm), which is adequate for measuring fluorophores with subnanosecond lifetimes. To characterize the system, NIR dyes were deposited directly on different substrate materials typically used for DNA microarrays, and the fluorescence lifetimes of two representative dyes were measured. The fluorescence lifetime for aluminum tetrasulfonated naphthalocyanine was found to be 1.92 ns, and a value of 1.21 ns was determined for the tricarbocyanine dye, IRD800, when it was deposited onto poly(methyl methacrylate) (PMMA) and measured in the dry state. Finally, the imager was used to monitor hybridization events using probe oligonucleotides chemically tethered to a PMMA substrate via a glutardialdehyde linkage to an aminated-PMMA surface. The limit of detection for oligonucleotides containing a NIR fluorescent reporter was determined to be 0.38 molecules/microm2, with this detection limit improving by a factor of 10 when a time-gate was implemented. Fluorescence lifetime analysis of the hybridization events on PMMA indicated a lifetime value of 1.23 ns for the NIR-labeled oligonucleotides when using maximum-likelihood estimators.     

Liquid sample injection using an atmospheric pressure direct current glow discharge ionization source.

An atmospheric pressure DC glow discharge in helium has been used as an ionization source for organic samples introduced by liquid injection into atmospheric pressure ionization mass spectrometry (API/MS). The glow source operates typically in the range up to 1 mA of current at less than 1 kV, although the source can be operated up to a discharge current of 10 mA. Even at the high current used in this work, the protonated molecule, MH+, is observed with little or no fragmentation for many of the samples studied. The detection limits achieved for API glow discharge detection are typically in the low femtomole region for small organic molecules including small biological neurotransmitters, drugs, pesticides, phenylthiohydantoin-substituted amino acids, and explosives. A detection limit of approximately 2 pg has been achieved for tyramine with linear quantitation over at least 3 orders of magnitude. The sensitivity in these experiments has been further improved by optimization of the skimmer-interface system and the liquid injection/nebulization design.     

Preparation of luminescent layered zirconium phosphate nanocomposites by the layer-by-layer technique

In the present work, photoactive cation N, N′-Dimenthyl-9, 9’-bisacridinium nitrate (BNMA) was assembled with exfoliated layered α-zirconium phosphate (α-ZrP) via an electrostatic layer-by-layer (LBL) assembly method. As a result, the luminescent films which were well-aligned and periodical had been successfully fabricated. Surprisingly, the lifetimes of (BNMA/ZrP)_n were found to be prolonged by 16-fold for the first time, due to the isolation effect of inorganic nanosheets and hydrogen ion migration between the interlayers. Therefore, it is testified that α-ZrP can be used as the laminate and has remarkable influences on enhancing the lifetimes of chromophores. We expect that this new discovered effect can enable α-ZrP a kind of new potential material to develop novel light-emitting materials and optical devices.     

Fluorescence lifetime correlation spectroscopic study of fluorophore-labeled silver nanoparticles

In this paper, we introduce the use of fluorescence lifetime correlation spectroscopy to study the metal-fluorophore interactions in solution at the single-fluorophore level. A single-stranded oligonucleotide was chemically bound to a 50-nm-diameter single silver particle, and a Cy5-labeled complementary single-stranded oligonucleotide was hybridized with the silver particle-bound oligonucleotide. The distance between the fluorophore and silver particle was maintained by a rigid hybridized DNA duplex of 8 nm in length. The single Cy5-DNA-Ag particles showed more than 10-fold increase in fluorescence intensity and a 5-fold decrease in emission lifetimes as compared with Cy5-DNA free molecules in the absence of metal. The decrease of lifetime for the Cy5-DNA-Ag particle allowed us to resolve the correlation functions of the two species based on the intensity decays. The increased brightness of the Cy5-DNA-Ag particle as compared to free Cy5-DNA resulted in an increased contribution of Cy5-DNA-Ag to the correlation function of the mixture. These results show that the effects of metal particles on fluorophores can be used to detect the small fractional populations of the metal-bound species in the presence of a larger number of less bright species. Our results also suggest that these bright fluorophores conjugated to silver particles could be used as the fluorescent probes for clinical detection in the biological samples with the high background.     

The halocarbon contamination problem. Part I: Atmospheric effects of halocarbons

The remarkable chemical and biological stability of many halocarbons, which makes them ideal refrigerants and aerosol propellants, has led to their accumulation in the lower atmosphere. In 1974, Rowland and Molina pointed out that these substances would eventually travel up to the stratosphere where they decompose to yield free chlorine atoms which catalytically destroy ozone. Continued release of F11 and F12 at 1973 production rates was calculated to produce an eventual ozone depletion of the order of 10%. Much subsequent research has not modified this estimate to any large extent but has emphasized the considerable uncertainties of such estimates. These uncertainties, and particularly the differences between calculated and measured concentrations of chlorine species in the atmosphere, wil be considered. Other topics to be discussed are the difficulties of detecting ozone depletions and the atmospheric lifetimes of plausible substitutes as refrigerants for the fully halogenated methanes and ethanes. Recent developments have modified our views on both these topics.     

Capillary electrophoresis in 2 and 5 microns diameter capillaries: application to cytoplasmic analysis

Capillary electrophoresis with electrochemical detection in 2- and 5-micron capillaries has been developed to study ultrasmall biological environments. Sample volumes as low as 270 fL have been injected into the electrophoresis capillary with subattomole detection limits for easily oxidized species. We have applied this method to the analysis of single cell cytoplasm. Sampling of the cytoplasm is accomplished by inserting one end of the electrophoresis capillary directly into a single nerve cell. The high-voltage end of the electrophoresis capillary has been etched with hydrofluoric acid to form a microinjector. This injection scheme represents an improvement over those previously used for similar applications. The excellent selectivity of this method is demonstrated for catecholamine and indolamine neurotransmitters and their metabolites found in the invertebrate system, the pond snail Planorbis corneus.     

Mechanistic studies of photochemical reactions with millisecond time resolution by electrospray ionization mass spectrometry

Electrospray ionization mass spectrometry is used as an on-line technique to investigate the solution photochemistry of [CpFebz]+ (Cp, eta5-cyclopentadienyl; bz, eta6-benzene). Direct irradiation of samples in the optically transparent tip of the electrospray source allows the detection of products with lifetimes down to the millisecond regime. Photolysis of [CpFebz]+ in acetonitrile (AN) yields the short-lived half-sandwich complex, [CpFe(AN)3]+. The suspected parent-offspring relationship between this species and the fully ring-deligated product, [Fe(AN)6]2+, has been confirmed. In solutions containing cyclohexene oxide, [CpFebz]+ generates several photoproducts capable of initiating the cationic polymerization of the epoxide monomer. The initiation process and the first few steps in the subsequent growth of the polymer chain have been observed.     

Mounting stripper foils on forks for maximum lifetime

While research and development continue to produce forms of carbon for longer lasting stripper foils, relatively little attention has been paid to other factors that affect their survival in use. It becomes apparent that the form of carbon is only part of the issue. Specific mounting methods increase the lifetimes of carbon stripper foils. These methods are determined in part by the specific use and carbon type for a foil. With careful handling, appropriate adhesive, and slack mounting, premature breakage can be avoided. Foil lifetimes are then primarily affected by less easily controlled factors such as high-temperature expansion, shrinkage and evaporation.     

Design and performance of capping layers for extreme-ultraviolet multilayer mirrors

Multilayer lifetime has emerged as one of the major issues for the commercialization of extreme-ultraviolet lithography (EUVL). We describe the performance of an oxidation-resistant capping layer of Ru atop multilayers that results in a reflectivity above 69% at 13.2 nm, which is suitable for EUVL projection optics and has been tested with accelerated electron-beam and extreme-ultraviolet (EUV) light in a water-vapor environment. Based on accelerated exposure results, we calculated multilayer lifetimes for all reflective mirrors in a typical commercial EUVL tool and concluded that Ru-capped multilayers have approximately 40x longer lifetimes than Si-capped multilayers, which translates to 3 months to many years, depending on the mirror dose.     

Estimation of atmospheric lifetimes of hydrofluorocarbons, hydrofluoroethers, and olefins by chlorine photolysis using gas-phase NMR spectroscopy

An empirical correlation has been derived between accepted atmospheric lifetimes of a set of hydrofluorocarbons and hydrofluoroethers and relative rates of reaction with photolyzed chlorine in excess at ambient temperature. These kinetic systems were studied by nuclear magnetic resonance (NMR) spectroscopy in the gas phase, marking the first application of NMR spectroscopy to this field. The square of the Pearson coefficient R for the linear correlation between observed reaction rates and accepted atmospheric lifetimes was 0.87 for compounds of lifetime less than 20 years. The method was extended to the study of ethene and propene; the rate of reaction of propene was found to be 1.25 times that of ethene at 23 degrees C. The chief advantage of this method is its simplicity and reliance only on common tools and techniques of an industrial chemical laboratory.     

Recent Progress of Singlet‐Exciton‐Harvesting Fluorescent Organic Light‐Emitting Diodes by Energy Transfer Processes

The external quantum efficiency (EQE) of organic light‐emitting diodes (OLEDs) has been dramatically improved by developing highly efficient organic emitters such as phosphorescent emitters and thermally activated delayed fluorescent (TADF) emitters. However, high‐EQE OLED technologies suffer from relatively poor device lifetimes in spite of their high EQEs. In particular, the short lifetimes of blue phosphorescent and TADF OLEDs remain a big hurdle to overcome. Therefore, the high‐EQE approach harvesting singlet excitons of fluorescent emitters by energy transfer processes from the host or sensitizer has been explored as an alternative for high‐EQE OLED strategies. Recently, there has been a big jump in the EQE and device lifetime of singlet‐exciton‐harvesting fluorescent OLEDs. Recent progress on the materials and device structure is discussed herein.