Laser-Based Fluorescence Eem Instrument for In-Situ Groundwater Monitoring

I. ABSTRACT

A fluorescence excitation-emission matrix (EEM) instrument is described which employs laser excitation and optical fiber light delivery for remote sensing applications. Ultraviolet laser light is generated at a variety of discrete wavelengths by pumping a Raman shifter with the third or fourth harmonic of a Nd:YAG laser, and this Raman-shifted light is used for fluorescence excitation to generate EEM spectra. Seven-meter optical fibers are used for laser beam delivery and fluorescence light collection, and a diode array detector is used to detect fluorescence emission spectra. As a test of the capability of this instrument, the composition of a two-component mixture of the groundwater pollutants carbazole and p-cresol is determined quantitatively by least-squares analysis of the mixture's fluorescence EEM. Detection limits were in the ppb range and the linear dynamic range was better than 3 orders of magnitude.     

In vivo imaging of cellular structures in Caenorhabditis elegans by combined TPEF, SHG and THG microscopy

In this study, we use combined two‐photon excitation fluorescence (TPEF), second‐harmonic generation (SHG) and third‐harmonic generation (THG) measurements to image cellular structures of the nematode Caenorhabditis elegans, in vivo. To our knowledge, this is the first time that a THG modality is employed to image live C. elegans specimens. Femtosecond laser pulses (1028 nm) were utilized for excitation. Detailed and specific structural and anatomical features can be visualized, by recording THG signals. Thus, the combination of three image‐contrast modes (TPEF‐SHG‐THG) in a single instrument has the potential to provide unique and complementary information about the structure and function of tissues and individual cells of live biological specimens.     

Controlled Synthesis and Optical Properties of Pure Gold Nanoparticles

Abstract

Gold nanoparticles were synthesized by laser ablation of a gold metallic disc at wavelengths of 532 nm and 355 nm with 7 ns pulse duration in the pure water. The colloidal gold nanoparticles were characterized by ultraviolet-visible absorption spectroscopy, transmission electron microscopy, and fluorescence spectrometry. The presence of a surface plasmon resonance peak around ～ 524 nm indicates the formation of gold nanoparticles. The formation efficiencies of gold nanoparticles in colloids were found to increase when ablating the gold metallic disc with a laser having a longer wavelength. The size distributions of the gold nanoparticles thus produced were measured by transmission electron microscopy. A reduction in mean diameter of the particles was observed with a decrease in the laser wavelength under the irradiation at a high fluence of 25 mJ/pulse. The fluorescence spectroscopy demonstrated that these gold nanoparticles are fluorescent, showing a strong blue emission intensity at 458 nm.     

Optical transition process in AgOx super-resolution near-field structure

A sandwiched 15 nm AgO_x thin film of the super‐resolution, near‐field optical disk was studied using a confocal Z‐scan system. Nonlinear optical properties of quartz glass/ZnS–SiO₂ (170 nm)/AgO_x (15 nm)/ZnS–SiO₂ (40 nm) were measured using a Q‐switch Nd : YAG pulse laser of wavelength 532 nm, pulse width 0.7 ns, and 15.79 kHz repetition rate. Transmittance and reflectance of the sandwiched AgO_x thin film show important optical responses at the focused position of Z‐scan. The dissociation processes of AgO_x, recombination of the silver and oxygen, and the resonance of the localized surface plasmon of the nano‐composites of the AgO_x thin film are correlated to transmittance and reflectance at the focused position of the Z‐scan for different input laser powers. An irreversible upper threshold intensity of 4.40 × 10⁶ mW cm^?2 at the focused position was found. A reversible working window of the focusing intensity between 1.86 × 10⁶ and 4.40 × 10⁶ mW cm^?2 was measured with sandwiched AgO_x thin film alone. The near‐field interactions of the AgO_x thin film and the recording layers of super‐resolution near‐field optical disk are also discussed.     

Application of blue laser direct-writing equipment for manufacturing of periodic and aperiodic nanostructure patterns

This study presents the novel development of low cost, highly efficient blue laser direct-writing equipment for using mask-less laser lithography to manufacture periodic and aperiodic nanostructure patterns. The system includes a long-stroke linear motor precision stage (X, Y), a piezoelectric nano-precision stage (Y, θz), a 3-DOF (degrees of freedom) laser interferometer measurement system, and a blue laser direct-writing optical system. The 3-DOF laser interferometer measurement system gives the control system feedback for displacement (X, Y, θz) of the equipment. The laser processing equipment consists of a blue laser direct-writing optical head, a field-programmable gate array (FPGA) alignment interface, and an optical head servo controller. The optical head operates at a wavelength of 405 nm. Processing the nanostructures on thermo-reaction inorganic resists with precise control of the laser intensity, taking advantage of the threshold effect to exceed the limitations of optical diffraction, and reduces the nanostructure hole size. The equipment can be used to fabricate various periodic nanostructure patterns, aperiodic nanostructure patterns, and two-dimensional patterns. The equipment positioning accuracy is within 50 nm at a speed of 50 mm/s, and the minimum critical dimension can be achieved about 100 nm or so.     

Plasmonic effect of silver nanoparticles on the upconversion emissions of Sm3+-doped sodium-borosilicate glass

The effect of the localized surface plasmon resonance (SPR) on optical absorption and photoluminescence of Sm³⁺-doped sodium borosilicate glass containing reduced silver nanoparticles (NPs) is reported (Ag⁺ → Ag⁰). The interaction of ultraviolet light by metallic NPs and its effect on the optical properties of samarium in proposed glass were investigated by absorption and photoluminescence spectra analysis. The existence of the NPs was pursued by transmission electron microscopy technique, revealing the existence of Ag NPs with average size of ∼8–14 nm. The largest enhancement was achieved for emission at 561 nm. Such improvements were attributed and discussed by enhanced electric field around metallic NPs and energy transfer (ET) between Sm³⁺ ions and silver NPs.     

PHOEBE,a prototype scanning laser-feedback microscope for imaging biological cells in aqueous media

Based on the principle of laser-feedback interferometry (LFI), a laser-feedback microscope (LFM) has been constructed capable of providing an axial (z) resolution of a target surface topography of ～ 1 nm and a lateral (x, y) resolution of ～ 200 nm when used with a high-numerical-aperture oil-immersion microscope objective. LFI is a form of interferometry in which a laser's intensity is modulated by light re-entering the illuminating laser. Interfering with the light circulating in the laser resonant cavity, this back-reflected light gives information about an object's position and reflectivity. Using a 1-mW He–Ne (λ = 632·8 nm) laser, this microscope (PHOEBE) is capable of obtaining 256 × 256-pixel images over fields from (10 μm × 10 μm) to (120 μm × 120 μm) in ～ 30 s. An electromechanical feedback circuit holds the optical pathlength between the laser output mirror and a point on the scanned object constant; this allows two types of images (surface topography and surface reflectivity) to be obtained simultaneously. For biological cells, imaging can be accomplished using back-reflected light originating from small refractive-index changes (> 0·02) at cell membrane/water interfaces; alternatively, the optical pathlength through the cell interior can be measured point-by-point by growing or placing a cell suspension on a higher-reflecting substrate (glass or a silicon wafer). Advantages of the laser-feedback microscope in comparison to other confocal optical microscopes include: the simplicity of the single-axis interferometric design; the confocal property of the laser-feedback microscope (a virtual pinhole), which is achieved by the requirement that only light that re-enters the laser meeting the stringent frequency, spatial (TEM₀₀), and coherence requirements of the laser cavity resonator mode modulate the laser intensity; and the improved axial resolution, which is based on interferometric measurement of optical amplitude and phase rather than by use of a pinhole as in other types of confocal microscopes.     

A laser desorption ion-mobility increment spectrometer for detection of ultralow concentrations of nitro compounds

A compact instrument that implements for the first time a method for analyzing substances, in which laser desorption of analyzed molecules and laser ionization of an air sample are combined with the ionmobility increment spectroscopy, is described. Pulse radiation of the fourth harmonic of a portable (2.6 kg) GSGG: Cr³⁺: Nd³⁺ laser (λ = 266 nm) is used. The detection limit of the developed laser desorption spectrometer of the ion-mobility increment is 40 pg for trinitrotoluene (TNT), and the linear dynamic range for TNT is 0.1–20.0 ng. The results from detection of nitro compounds are presented: trinitrotoluene, cyclotrimethylene trinitramine (RDX), and octogen (HMX). It is shown that laser desorption of nitro compounds from a metal is accompanied by their decomposition on the surface and emission of fragments. An ion signal is obtained for nitro compounds that were ionized outside the spectrometer.     

The use of optical parametric oscillator for harmonic generation and two-photon UV fluorescence microscopy

Ultrafast lasers have found increasing use in scanning optical microscopy due to their very high peak power in generating multiphoton excitations. A mode-locked Ti:sapphire laser is often employed for such purposes. Together with a synchronously pumped optical parametric oscillator (OPO), the spectral range available can be extended to 1,050-1,300 nm. This broader range available greatly facilitates the excitation of second harmonic generation (SHG) and third harmonic generation (THG) due to better satisfaction of phase matching condition that is achieved with a longer excitation wavelength. Dental sections are then investigated with the contrasts from harmonic generation. In addition, through intra-cavity doubling wavelengths from 525-650 nm are made available for effective two-photon (2-p) excitation with the equivalent photon energy in the UVB range (290-320 nm) and beyond. This new capacity allows UV (auto-) fluorescence excitation and imaging, for example, from some amino acids, such as tyrosine, tryptophan, and glycine.     

Deuterium Lamp Power Supply

Abstract

The development of a near-infrared fiber optic biomolecular probe (FOBP) for the detection of biological molecules will be discussed. The utility of the system is demonstrated using an immunochemical samples. The FOBP consists of a semiconductor laser diode (γ_max = 780 nm), a modified fiber tip binding site, an NIR dye and a detector. The common distal end of two optical plastic fibers, connected with poly(methyl methacrylate), serves as the binding site (about < 2mm^X in diameter). Activation of the binding site with different inorganic acids is discussed. Preparation of the FOBP involved in the immobilization anti-IgG antibody followed by coating with IgG for a sandwich type probe. Immobilization of anti-IgG antibody was achieved by reacting the Fc fragment of the antibody molecule to the activated binding site of the fiber. The instrument was tested by labeling the antibody with a commercially available near infrared (NIR) dye IR-144). Excitation of the antigen-antibody sandwich complex was induced with a semiconductor laser diode at 780 nm. The emission intensity signal was collected at 820 nm by using small bandwidth filters. The response time for 95% of the signal was less than 5 seconds. The low background signal of the detector allowed the detection of 2.72 ng/mL IgG in a probe coated with 10 ng/mL antibody. The general procedure used for the attachment of the biological molecule to the distal end of the probe can be used to construct more specific antigen-antibody systems. Use of smaller components shown here will allow for the manufacturing of portable instruments for applications in medicine and clinical analysis.     

Two-photon fluorescence surface wave microscopy

This paper demonstrates the principle of two-photon surface wave microscopy with a view to applications on biological samples. We describe a modified scanning optical microscope, which uses specially prepared coverslips. These coverslips are designed to support the propagation of surface waves capable of large field enhancements. We also discuss the beam conditioning necessary to ensure efficient use of the available illumination. Two-photon surface wave fluorescent excitation is demonstrated on fluorescent nanospheres, demonstrating a point spread function width of ≈220 nm at an illumination wavelength of 925 nm. The potential of non-linear surface wave excitation for both fluorescence and harmonic imaging microscopy is discussed.     

GaAs纳米线阵列光阴极的制备及其特性研究

采用改进的Stber法合成直径为500nm的SiO_2纳米球,用旋涂法将所合成的SiO_2纳米球制作掩模版,采用纳米球刻蚀法制备GaAs纳米线阵列。利用扫描电子显微镜(SEM)、漫反射谱对其进行了表征分析,再对所制备出的GaAs纳米线阵列结构进行Cs-F交替激活实验,使其表面形成负电子亲和势光阴极,并对最终制备出的GaAs纳米线阵列光阴极样品进行量子效率测试,验证了GaAs纳米线阵列结构的量子效率比GaAs基片提高50%以上,从而证实了纳米线阵列结构的高光电转换效率。     

Effect of nickel nanopowder on lubrication behaviour of low-temperature grease in steel-steel tribosystem

ABSTRACT

Nickel nanoparticles with sizes from 12 to 34?nm were obtained by pyrolysis of nickel formate. They have been used as additives to the low-temperature grease (CIATIM-201). Studies of tribotechnical properties have shown that the CIATIM-201 with nickel additives at concentrations of 0.05 wt. % - 0.2 wt. % has the best antiwear and extreme pressure properties in comparison with pure grease. Friction surfaces were characterised by the methods of atomic-force microscopy AFM (PHYWE), 3D optical microscopy and indentation tests. The improvement of tribotechnical properties of grease is related to the process of micro-polishing, which leads to the formation of a structural layer with a high modulus of elasticity on the friction surface.     

Optical Fiber Metal Probes Using Semiconductor Laser Diodes and Near-Infrared Compounds

Abstract

The development of a near-infrared optical fiber detector for the determination of metal ions was investigated. The optical fiber detector consists of a semiconductor laser diode, an optical fiber, an NIR dye and a photodiode detector. Two plastic fibers are connected together with poly(methyl methacrylate) (PMMA) with a reaction phase at the distal common end. The reaction phase had a diameter of about <2 mm. Excitation of the dye/analyte complex was induced with a semiconductor laser diode at 780 nm and the emission intensity signal was collected by two instrument systems. No photobleaching effect was observed during the investigations. The response time for 95% of the signal was less than 5 seconds. The use of plastic fibers has allowed easy immobilization of an NIR dye. The dye was entrapped in a polymer matrix that allows the diffusion of the analyte. Low background signal of the photodiode permitted the detection of dye in the picomolar level. Comparison of the optical fiber results between a PMT and a photodiode was made. The use of smaller components shown here will allow the manufacturing of portable optical fiber detectors for applications in industry, medicine and the environment.     

Multiphoton microscopy in life sciences

Near infrared (NIR) multiphoton microscopy is becoming a novel optical tool of choice for fluorescence imaging with high spatial and temporal resolution, diagnostics, photochemistry and nanoprocessing within living cells and tissues. Three‐dimensional fluorescence imaging based on non‐resonant two‐photon or three‐photon fluorophor excitation requires light intensities in the range of MW cm^?2 to GW cm^?2, which can be derived by diffraction limited focusing of continuous wave and pulsed NIR laser radiation. NIR lasers can be employed as the excitation source for multifluorophor multiphoton excitation and hence multicolour imaging. In combination with fluorescence in situ hybridization (FISH), this novel approach can be used for multi‐gene detection (multiphoton multicolour FISH). Owing to the high NIR penetration depth, non‐invasive optical biopsies can be obtained from patients and ex vivo tissue by morphological and functional fluorescence imaging of endogenous fluorophores such as NAD(P)H, flavin, lipofuscin, porphyrins, collagen and elastin. Recent botanical applications of multiphoton microscopy include depth‐resolved imaging of pigments (chlorophyll) and green fluorescent proteins as well as non‐invasive fluorophore loading into single living plant cells. Non‐destructive fluorescence imaging with multiphoton microscopes is limited to an optical window. Above certain intensities, multiphoton laser microscopy leads to impaired cellular reproduction, formation of giant cells, oxidative stress and apoptosis‐like cell death. Major intracellular targets of photodamage in animal cells are mitochondria as well as the Golgi apparatus. The damage is most likely based on a two‐photon excitation process rather than a one‐photon or three‐photon event. Picosecond and femtosecond laser microscopes therefore provide approximately the same safe relative optical window for two‐photon vital cell studies. In labelled cells, additional phototoxic effects may occur via photodynamic action. This has been demonstrated for aminolevulinic acid‐induced protoporphyrin IX and other porphyrin sensitizers in cells. When the light intensity in NIR microscopes is increased to TW cm^?2 levels, highly localized optical breakdown and plasma formation do occur. These femtosecond NIR laser microscopes can also be used as novel ultraprecise nanosurgical tools with cut sizes between 100 nm and 300 nm. Using the versatile nanoscalpel, intracellular dissection of chromosomes within living cells can be performed without perturbing the outer cell membrane. Moreover, cells remain alive. Non‐invasive NIR laser surgery within a living cell or within an organelle is therefore possible.     

A laser spectrometer of field-asymmetric ion mobility

A method for analyzing a substance has been experimentally tested. The method combines the field-asymmetric ion mobility spectrometry and laser ionization of molecules under atmospheric pressure. Pulsed radiation of the fourth harmonic of an YAG: Nd³⁺ laser (λ = 266 nm) and a spectrometer with a cylindrical analysis camera were used. The results of detecting nitrocompounds—trinitrotoluene, cyclotrimethylenetrinitramine (hexogen, RDX), etc.—are presented. The experimental detection limits of the spectrometer are 5 × 10⁻¹⁵ g/cm³ (cyclotrimethylenetrinitramine) and ≤3 × 10⁻¹⁵ g/cm³ (trinitrotoluene).     

全固态589 nm复合腔连续波和频激光器

给出了一种复合腔结构和频激光器,用2台激光二极管阵列(LDA)经过光纤耦合分别单独端面抽运Nd:YVO₄和Nd:YAG晶体,其中Nd:YVO₄和Nd:YAG晶体所选择的能级跃迁分别为⁴F_3/2-⁴I_11/2和⁴F_3/2-⁴I_13/2,其对应激光跃迁波长分别为1 064 nm和1 319 nm,两基频激光束分别在两个子谐振腔中振荡,在其交叠区利用KTP II类临界相位匹配(CPM)进行腔内和频,获得了589 nm的和频激光。当抽运功率为8 W/14 W时获得了340 mW连续波TEM₀₀黄激光输出。光束质量因子M²<1.2,激光输出功率噪声低,4 h功率不稳定度小于±3％。该复合腔结构是实现LDA泵浦589 nm全固态黄光激光器一种有效的和频方法。     

A Multifrequency Phase Fluorometer Using the Harmonic Content of a Mode-Locked Laser

ABSTRACT

We describe the construction and operation of a cross-correlation phase and modulation fluorometer which uses the harmonic content of a high repetition rate mode-locked laser as the excitation source.

A mode-locked argon ion laser is used to synchronously pump a dye laser. The pulse train output from the dye laser is amplitude modulated by an acousto-optic modulator and then frequency doubled with an angle tuned frequency doubler. With the particular dye utilized in these studies, the ultraviolet light obtained was continuously tunable over the range 280-310 nm. In the frequency domain the high repetition rate pulsed source gives a large series of equally spaced harmonic frequencies. The frequency spacing of the harmonics is determined by the repetition frequency of the laser. Amplitude modulation of the pulse train permits variation of the frequency quasi-continuously from a few hertz to gigahertz. Use of cross-correlation techniques permits precise isolation of individual frequencies. The cross-correlation frequency required for the analysis of the phase delay and modulation ratio is obtained using coupled frequency synthesizers. In the present instrument three synthesizers are used. One drives the pump mode-locker head, a second drives the acousto-optic modulator and the third is used to modulate the response of the photomultiplier tubes which detect the signal. The accuracy, reproducibility and sensitivity of the instrumentation have been determined. Experimental data are provided to show use of this high frequency cross-correlation phase-modulation fluorometer for the determination of fluorescence lifetimes and rotational motions of tryptophan in solution and in proteins.     

A high-energy YAG:Nd<Superscript>3+</Superscript> laser with fiber destroyed radiation coherence

The optical scheme and the design of a high-power YAG:Nd³⁺ laser, which generates radiation pulses of nanosecond duration at wavelengths of 1064 nm (0.8 J), 532 nm (160 mJ), and 266 nm (40 mJ) with destroyed coherence in a fiber and a smoothed intensity distribution in the beam cross section, are described. The processes that affect the generation efficiency of decoherenized radiation are considered.     

Detection in near-field domain of biomolecules adsorbed on a single metallic nanoparticle

In this study, neurodegeneration phenomena were investigated, by performing third harmonic generation imaging measurements on the nematode Caenorhabditis elegans, in vivo. The in vivo, precise identification of the contour of the degenerating neurons in the posterior part of the nematode and the monitoring, in real time, of the progression of degeneration in the worm, through third harmonic generation imaging measurements, were achieved. Femtosecond laser pulses (1028 nm) were utilized for excitation. Thus, the THG image contrast modality comprises a powerful diagnostic tool, providing valuable information and offering new insights into morphological changes and complex developmental processes in live biological specimens.