Laser induced fluorescence diagnostic for velocity distribution functions:applications,physics,methods and developments

With more than 30 years of development,laser-induced fluorescence(LIF)is becoming an increasingly common diagnostic to measure ion and neutral velocity distribution functions in different fields of studies in plasma science including Hall thrusters,linear devices,plasma processing,and basic plasma physical processes.In this paper,technical methods used in the LIF diagnostic,including modulation,collection optics,and wavelength calibration techniques are reviewed in detail.A few basic physical processes along with applications and future development associated with the LIF diagnostics are also reviewed.

Quantitative measurement of hydroxyl radical(OH) concentration in premixed flat flame by combining laser-induced fluorescence and direct absorption spectroscopy

An accurate and reasonable technique combining direct absorption spectroscopy and laser-induced fluorescence（LIF）methods is developed to quantitatively measure the concentrations of hydroxyl in CH;/air flat laminar flame. In our approach, particular attention is paid to the linear laser-induced fluorescence and absorption processes, and experimental details as well. Through measuring the temperature, LIF signal distribution and integrated absorption, spatially absolute OH concentrations profiles are successfully resolved. These experimental results are then compared with the numerical simulation. It is proved that the good quality of the results implies that this method is suitable for calibrating the OH-PLIF measurement in a practical combustor.

Quantitative measurements of one-dimensional OH absolute concentration profiles in a methane/air flat flame by bi-directional laser-induced fluorescence

The one-dimensional （1D） spatial distributions of OH absolute concentration in methane/air laminar premixed flat flame under different equivalence ratios at atmospheric pressure are investigated by using bi-directional laser-induced flu- orescence （LIF） detection scheme combined with the direct absorption spectroscopy. The effective peak absorption cross section and the average temperature at a height of 2 mm above the burner are obtained by exciting absorption on the Q1（8） rotational line in the A2∑＋ （Dt = 0） ←- X2∏ （v = 0） at 309.240 nm. The measured values are 1.86×10-15 cm2 and 1719 K, respectively. Spatial filtering and frequency filtering methods of reducing noise are used to deal with the experi- mental data, and the smoothing effects are also compared using the two methods. The spatial distribution regularities of OH concentration are obtained with the equivalence ratios ranging from 0.8 to 1.3. The spatial resolution of the measured result is 84μm. Finally, a comparison is made between the experimental result of this paper and other relevant study results.

Numerical analysis of quantitative measurement of hydroxyl radical concentration using laser-induced fluorescence in flame

The aim of the present work is to quantitatively measure the hydroxyl radical concentration by using LIF（laserinduced fluorescence） in flame.The detailed physical models of spectral absorption lineshape broadening,collisional transition and quenching at elevated pressure are built.The fine energy level structure of the OH molecule is illustrated to understand the process with laser-induced fluorescence emission and others in the case without radiation,which include collisional quenching,rotational energy transfer（RET）,and vibrational energy transfer（VET）.Based on these,some numerical results are achieved by simulations in order to evaluate the fluorescence yield at elevated pressure.These results are useful for understanding the real physical processes in OH-LIF technique and finding a way to calibrate the signal for quantitative measurement of OH concentration in a practical combustor.

Ion Transport Measurements in a Multi-Dipole Argon Plasma by Broadband Laser Induced Fluorescence

Argon ion laser induced fluorescence measurements were carried out in a multipolar filament discharge with a broadband diode laser centered on 668 nm, which stimulated a transition from the metastable state in Ar（Ⅲ） 3d4F7/2 to 4p4D0 5/2. The intensity of the induced fluorescence at 442 nm was maximized by the optimization of the discharge parameters and the laser power. From the recovery of the background fluorescence after the laser was turned off, the ion diffusion coefficient was deduced and compared with the result inferred from the experiments of ion acoustic wave （IAW） damping.

Detection of Petroleum Pollutants by Using Laser Induced Fluorescence Imaging

A new laser induced fluorescence imaging system used for detecting and locating the petroleum pollutants is presented. A unique feature of the system was using a combination of a frequency tripled Nd:YAG pulse laser and an image intensified CCD camera as imaging detector. The delay time of the system and the exposure time of the CCD are controlled by a delay generator. Petroleum pollutants on various backgrounds had been detected and located by this system. This imaging technique provides a useful noninvasive approach for the detection of petroleum pollutants, which can be used in remote sensing.

Absolute density measurement of nitrogen dioxide with cavity-enhanced laser-induced fluorescence

The absolute number density of nitrogen dioxide（NO2） seeded in argon is measured with cavity-enhanced laserinduced fluorescence（CELIF） through using a pulsed laser beam for the first time. The cavity ring down（CRD） signal is acquired simultaneously and used for normalizing the LIF signal and determining the relationship between the measured CELIF signal and the NO2 number density. The minimum detectable NO2 density down to（3.6±0.1）10^8 cm^-3 is measured in 60 s of acquisition time by the CELIF method. The minimum absorption coefficient is measured to be（2.0±0.1）10^-9 cm^-1, corresponding to a noise equivalent absorption sensitivity of（2.2±0.1）10^9 cm.^-1Hz^-1/2. The experimental system demonstrated here can be further improved in its sensitivity and used for environmental monitoring of outdoor NO2 pollution.

Simultaneous laser-induced fluorescence and contactless-conductivity detection for microfluidic chip

A combined detection system involving simultaneous LIF and contacfless-conductometric measurements at the same place of the microfluidic chip was described. The LIF measurement was designed according to the confocal principle and a moveable contactless-conduetivity detector was used in C^4D. Both measurements were mutually independent and advantageous in analyses of mixtures. Various experimental parameters affecting the response were examined and optimized. The performances were demonstrated by simultaneous detection of Rhodamine B. And the results showed that the combined detection system could be used sensitively and reliably.

Hyperspectral imaging and remote trace detection of cis-1,3,4,6-tetranitrooctahydroimidazo-[4,5 d]imidazole(BCHMX)compared with traditional explosives using laser induced fluorescence

cis-1,3,4,6-Tetranitrooctahydroimidazo-[4,5 d]imidazole(BCHMX)is an advanced energetic compound that expected to spread worldwide in the near future.Since,no approved remote detection methods were reported in current literature for this material,we performed hyper-spectral imaging and laser induced fluorescence(LIF)to a BCHMX sample under low laser fluence for determining the optimum laser wavelength used in any future BCHMX-LIF based remote detection systems.For this purpose,an experimental setup consisted of a sun spectrum lamp and hyper-spectral camera was built to illuminate and image white powder samples of BCHMX in comparison with the traditional explosives,HMX(1,3,5,7-tetranitro-1,3,5,7-tetraazacyclooctane),RDX(1,3,5-trinitro-1,3,5-triazacyclohexane),PETN(2,2-Bis[(nitroxy)methyl]propane-1,3-diyldinitrate).The imaging reveals strong BCHMX sample absorption contrast among other samples at wavelength ranging from 400 to 410 nm.When light source was replaced by a 405 nm laser diode illuminator,a strong BCHMX sample LIF at the spectral range from 425 to 700 nm was observed under low laser fluence condition of 0.1 mJ/cm^(2).Finally,we demonstrated successfully the ability of the 405 nm LIF and the hyperspectral imaging technique to detect finger print traces of BCHMX on white cellulose fabric from a distance of 15 m and a detection limit of 1 mg/cm^(2).

Laser Induced Fluorescence Spectroscopy of IrN

High resolution laser induced fluorescence, spectra of IrN in the spectral region between 394 and 520 nm were recorded using laser vaporization/reaction free jet expansion and laser induced fluorescence spectroscopy. Seven new vibronic transition bands were observed and analyzed. Two Ω=1 and five Ω=0 new states were identified. Least squares fit of rotationally resolved transition lines yielded accurate molecular constants for the upper states. Spectra of isotopic molecules were observed, which provided confirmation for the vibrational assignment. Comparison of the observed electronic states of IrB, IrC, and IrN provides a good understanding of the chemical bonding of this group of molecules.

Laser scanning fluorescence microscopic measurement of the movement of cleaving egg surface of Rana Amurensis

By laser scanning fluorescence microscopy for quan-titative measurement of fluorescence intensity changes on egg surface stained with fluorescein isothiocyanate duxing cleavage furrow extending forward, it was found that in area of presumptive cleavage furrow the scanning curve became ∨ shape, indicating dark stripe appeared in that place. Then the fluorescence intensity increased at the place where the botton of ∨ shape had located, and the scanning curve tuxned to ∧ shape, indicating single stripe was formed. While enhanced fluorescence appeared on the borders of ∧ shape, an M shape curve was found, show-ing double stripe occurred. During the distance between two borders of M shape incresing from 50 μm to 100μm,a fluorescence peak came to sight in the middle of the M shape, which being the cleavge furrow bottom. The two lateral sides of furrow bottom with decreasing fluorescence were nascent membrane. At that time the curve became W shape. By the sides of cleavage furrow the the stress folds became conspicous after double stripe stage, showing the stretching of the egg surface being increased. With our[31, 33] and others[32] reports that polylysine could induce the appearance of nascent membrane and phyto-hemagglutinins could decrease or prevent the appearance of nascent membrane, we believed the idea of Schroeder[25] that increasing mechanical stress could initiate nascent membrane formation and thought that the stress lay to the outsides of cleavage furrow.

Research of Confocal Laser Induced Fluorescence Detection System for Micro-fluidic Chip

The characteristics such as signal noise ratio（SNR） and sensitivity of the fluorescence detection system for micro-fluidic chip influence the performance of the whole system extremely. The confocal laser induced fluorescence detection system is presented. Based on the debugging of optical and circuit modules, the results of detecting the samples are given and analyzed theoretically, and the improved project is put forward.

Temperature-Dependent Fluorescence Properties and Diode-Pumped Deep Red Laser Performance of Pr:LiGdF4 Crystal

We demonstrate the temperature-dependent fluorescence properties of Pr：LiGdF4 crystal tor the first time, to the best of our knowledge, and its blue diode pumped cw red laser at 720 nm at room temperature. The fluorescence lifetime and polarized emission cross sections in the visible range are measured and calculated in a temperature range from 77K to 300 K, and the variations of the fluorescence lifetime and spectra are discovered. The reasons for these changes are explained accordingly. The output wavelength of the 720nm laser is first reported on the laser performance by using a fiber-coupled laser diode at the wavelength of 442nm as the pump source, and the maximum cw output power is about 303roW.

Determination of Amino Acids in an Individual Erythrocyteby Capillary Electrophoresis with Intracellular FITC-derivatization and Laser-induced Fluorescence Detection

A novel approach for analysis of amino acids in individual erythrocytes was established. In this method, the derivatization reagent was introduced into the living cells by electroporation. After derivatization, the amino acids in a single cell were determined by capillary electrophoresis with laser-induced fluorescence detection.

Production of Polyclonal Antibody of Morphine and Determination of Morphine in Urine by Capillary Electrophoresis Immunoassay with Laser-induced Fluorescence Detection

N-Conjugated antigen was synthesized and polyclonal antibody with high specificity was obtained from immunizing animals. With this polyclonal antibody, a rapid and efficient CEIA-LIF method was developed to determine the free morphine in urine of abusers. The detection limit was calculated to be 40 ng/mL. Simulated urine samples were analyzed with good recoveries, which showed the feasibility of its application in specific morphine determination in urine of morphine abusers.

Detennination of trace uranium in environment by laser induced fluorescence method

he analytical technique of trace uranium has been developed rapidly since Robbins firstreported the laser uranium analyzer in 1978. Because of the complicated operation and experimentalprocedure of the normal spectrophotometer, neutron activation and solid fluorescence, they havebeen replaced gradually by the laser induced fluoresccncc mcthod. According to high sensitivity andIow detection limit of the fluoreseenee, it can determine the trace uranium. The uranium content inthe lake water, underground water and rain etc. can be measured directly without pretreatment. Therange of the method for determining uranium is from 005 ppb to 20 ppb, and the detection limitis 0.05 ppb.

Effect of Temperature on Optical Properties of Vegetable Oils Using UV-Vis and Laser Fluorescence Spectroscopy

UV-Vis absorption and fluorescence spectroscopy are used to test the quality and changes in the composition of extra virgin olive oil (EVOO) and canola oil (CO) with temperature. The increase of temperature caused a change in the molecular structures of both types of oils seen as a gradual decrease of intensity amplitudes of absorption and fluorescence signals. A significant alteration occurred at ≈200&#176C where almost the main spectra of pheophytin-a, b, carotenoids, lutein and vitamin E in EVOO and linoleic acid and oleic acid in CO disappeared. An independent experiment showed the output of laser changes linearly with the input in oil at constant temperature (i.e., room temperature) where the transmission values of ≈33% and ≈75% are determined for EVOO and CO respectively. However, the transmission through a heated oil exhibited a non-linear behaviour which indicates the molecular optical response to thermal changes. The effect of storage time and adulteration of oils were also evaluated.

Comparison of three fluorescence labeling and tracking methods of endothelial progenitor cells in laser-injured retina

AIM： To compare three kinds of fluorescent probes for in vitro labeling and in vivo tracking of endothelial progenitor cells（EPCs） in a mouse model of laser-induced retinal injury.METHODS： EPCs were isolated from human umbilical cord blood mononuclear cells and labeled with three different fluorescent probes： 5-（and-6）-carboxyfluorescein diacetate succinimidyl ester（CFSE）, 1,1′-dilinoleyl-3,3,3′,3′-tetramethylindo-carbocyanine perchlorate linked acetylated low-density lipoprotein（Di I-Ac LDL）, and green fluorescent protein（GFP）. The fluorescent intensity of EPCs was examined by confocal microscopy. Survival rate of labeled EPCs was calculated with trypan blue staining, and their adhesive capability was assessed. A mouse model of retinal injury was induced by laser, and EPCs were injected into the vitreous cavity. Frozen section and fluorescein angiography on flat-mounted retinal samples was employed to track the labeled EPCs in vivo.RESULTS： EPCs labeled with CFSE and Di I-Ac LDL exhibited an intense green and red fluorescence at the beginning; the fluorescence intensity decreased gradually to 20.23% and 49.99% respectively, after 28 d. On the contrary, the florescent intensity of GFP-labeled EPCs increased in a time-dependent manner. All labeled EPCs showed normal morphology and no significant change in survival and adhesive capability. In the mouse model, transplantation of EPCs showed a protective effect against retinal injury. EPCs labeled with CFSE and Di I-Ac LDL were successfully tracked in mice during the development of retinal injury and repair; however, GFP-labeled EPCs were not detected in the laser-injured mouse retina.CONCLUSION： The three fluorescent markers used in this study have their own set of advantages and disadvantages. CFSE and Di I-Ac LDL are suitable for short-term EPClabeling, while GFP should be used for long-term labeling. The choice of fluorescent markers should be guided by the purpose of the study.

Mixed Micellar Electrokinetic Chromatographic Analysis of Colistin, Polypeptide Antibiotic, Using Laser-Induced Fluorescence Detection

The main goal of this work was to quantify the detection of colistin at low levels in urine samples through the practical application of mixed surfactant micellar electrokinetic chromatography–laser-induced fluorescence (MEKC-LIF) analysis method using its advantage of sensitivity and to examine direct injection of biological samples. Colistin (po- lymyxin E) has neither strong UV chromophore nor fluorophore. So, its assay for metabolism, pharmacokinetics studies for bioavailability and bioequivalence are difficult because of poor detectability. Therefore an enhanced UV or fluores-cence detection by chemical derivatization is required. MEKC-LIF method was proposed for colistin with a 488/520 nm argon-ion laser using a pre-CE derivatization with fluorescein isothiocyanate (FITC). Borate buffer was used as background buffer (BGB). The different parameters affecting the proposed derivatization reaction including concentration of the derivatizing reagent, reaction time and temperature were studied and optimized. The derivative was stable for up to 3 days. Different micelles (TX-100 and SDS) were examined as BGB additives separately but negative-charged mixed micelles (SDS/TX-100) were shown to be the best additive to BGB for the analysis of colistin particularly in human urine as they enhance both selectivity and sensitivity of the proposed method. BGB was used with pH 9.5, 10 kV, 8 s inj time, capillary length 75 cm × 75 μm ID (66 cm effective length), detection was LIF Ex 488 nm;Em 520 nm. The method was applied to colistin analysis in human urine and the recovery was > 98% (n = 5). LOD and LOQ in urine after pre-column derivatization using FITC were 100 and 250 ng/ml, respectively. Urine samples were analysed by direct injection without sample pre-treatment. The mechanism of enhancement of fluorescence of the derivative by surfactant was proposed.

Dynamic Non-Invasive Detection of NADH Based on Blood Flow-Mediated Skin Fluorescence (FMSF) Method

Nicotinamide adenine dinucleotide (NADH/NAD+) is involved in important biochemical reactions in human metabolism, including participation in energy production by mitochondria. The changes in fluorescence intensity as a function of time in response to blocking and releasing of blood flow in a forearm are used as a measure of oxygen transport with blood to the tissue, which directly correlates with the skin microcirculation status. In this paper, a non-invasive dynamic monitoring system based on blood flow-mediated skin fluorescence (FMSF) technology is developed to monitor the NADH fluorescence intensity of skin tissue during the process of blocking reactive hyperemia. Simultaneously, laser speckle contrast imaging (LSCI) and laser Doppler flowmetry (LDF) were used to observe blood flow, blood oxygen saturation (SOt2) and relative amount of hemoglobin (rHb) during the measurement process, which helped to explore NADH dynamics relevant physiological changes. A variety of parameters have been derived to describe NADH fluorescence curve based on the FMSF device. The experimental results are conducive to understanding the NADH measurement and the physiological processes related to it, which help FMSF to be a great avenue for in vivo physiological, clinical and pharmacological research on mitochondrial metabolism.