Integration of microfluidics with biomedical infrared spectroscopy for analytical and diagnostic metabolic profiling

We describe how infrared spectroscopy of dry films (IRDF) can provide diagnostic information, and how we expect integration with laminar fluid diffusion interface (LFDI) sample pre-processing to generate new analytical and diagnostic tests. LFDI pre-processing provides sample clean-up and analyte separation. The sensitivity of IRDF to certain analytes is enhanced through the depletion of sample constituents that otherwise obscure relevant spectral features, permitting the deposition of films with larger sample volumes and, hence, of greater effective optical pathlength for the targeted analytes. An integrated LFDI-IRDF technology holds promise both as a method for rapid point-of-care quantitative analysis of biological fluids and as the engine of discovery for a wide range of novel diagnostic methods based upon metabolic profiling. In particular, successful integration will provide a versatile and cost effective technology platform that will allow for the accurate quantification of low-concentration analytes that are otherwise inaccessible and will provide the basis for diagnostic and prognostic methods that would otherwise be impossible. The specific question addressed by the proof-of-concept study summarised here is whether the spectra of LFDI processed samples can provide analytical methods that are more accurate than otherwise possible without LFDI pre-processing. The enrichment of serum creatinine is accomplished, with subsequent enhancement of its spectral contribution permitting quantification of this clinically important analyte beyond that achievable with no pre-processing. Finally, to illustrate the potential in diagnostic applications, two recently initiated studies are outlined, one involving chronic kidney disease and the other for chronic and acute coronary artery disease.     

火花放电原子发射光谱法测定球化铁水中C,S元素的制样方法研究

用发射光谱仪精准测定球化铁水C元素一直是铸造行业的一大难题,由于样品制备误差具有显著性,测定用的科学仪器校准好,不等于测定的产品成分真实客观。国标GB/T 20066-2006《钢和铁化学成分测定用试样的取样和制样方法》对球化铁水测定未给出明确的制样方法,应用企业无法按统一方法进行生产过程控制,精准诊断产品化学成分可以降低可疑品数量和质量控制成本。本文介绍用发射光谱仪精准测定球化铁水化学成分样品制备方法认证,可以解决用发射光谱仪精准测定球化铁水化学成分技术难题。     

Atomic layer deposition of Al(2)O(3) and ZnO at atmospheric pressure in a flow tube reactor

Improving nanoscale thin film deposition techniques such as atomic layer deposition (ALD) to permit operation at ambient pressure is important for high-throughput roll-to-roll processing of emerging flexible substrates, including polymer sheets and textiles. We present and investigate a novel reactor design for inorganic materials growth by ALD at atmospheric pressure. The reactor uses a custom "pressure boost" approach for delivery of low vapor pressure ALD precursors that controls precursor dose independent of reactor pressure. Analysis of continuum gas flow in the reactor shows key relations among reactor pressure, inert gas flow rate, and species diffusion that define conditions needed to efficiently remove product and adsorbed reactive species from the substrate surface during the inert gas purge cycle. Experimental results, including in situ quartz crystal microbalance (QCM) characterization and film thickness measurements for deposition of ZnO and Al(2)O(3) are presented and analyzed as a function of pressure and gas flow rates at 100 °C. At atmospheric pressure and high gas flow, ZnO deposition can proceed at the same mass uptake and growth rate as observed during more typical low pressure ALD. However, under the same high pressure and flow conditions the mass uptake and growth rate for Al(2)O(3) is a factor of ～1.5-2 larger than at low pressure. Under these conditions, Al(2)O(3) growth at atmospheric pressure in a "flow-through" geometry on complex high surface area textile materials is sufficiently uniform to yield functional uniform coatings.     

Infrared multiphoton dissociation and electron-induced dissociation as alternative MS/MS strategies for metabolite identification

A major challenge encountered in mass spectrometric metabolite analysis is the identification and structural characterization of metabolites. Fourier transform ion cyclotron resonance mass spectrometry is a valuable technique for metabolite structural determination because it provides accurate masses and allows for multiple MS/MS fragmentation strategies, including infrared multiphoton dissociation (IRMPD) and electron-induced dissociation (EID). Collision activated dissociation (CAD) is currently the most commonly used MS/MS technique for metabolite structural characterization. In contrast, IRMPD and EID have had very limited, if any, application for metabolite characterization. Here, we explore IRMPD and EID of phosphate-containing metabolites and compare the resulting fragmentation patterns to those of CAD. Our results show that CAD, IRMPD, and EID provide complementary structural information for phosphate-containing metabolites. Overall, CAD provided the most extensive fragmentation for smaller (<600 Da) phosphate-containing metabolites; however, IRMPD generated more extensive fragmentation for larger (>600 Da) phosphate-containing metabolites, particularly for species containing increased numbers of phosphate groups. EID generally provided complementary fragmentation to CAD and showed extensive fragmentation with relatively evenly abundant product ions, regardless of metabolite size. However, EID fragmentation efficiency is lower than those of CAD and IRMPD.     

Physical and mechanical properties of reactively sputtered chromium boron nitride thin films

This paper reports on the first study of physical and mechanical properties of reactively sputtered chromium boron nitride coatings as a function of chemical composition, bias voltage and substrate temperature. Several sets of coatings were deposited by reactive unbalanced magnetron sputtering on Si(100) substrates. The chemical composition was deduced from X-ray photoelectron spectroscopy and Auger electron spectroscopy measurements, and was found to be influenced primarily by nitrogen flow rate. The phase composition was determined using X-ray diffraction in conjunction with spectroscopic ellipsometry. Atomic force microscopy was utilized to determine surface roughness and average surface grain size. Both surface roughness and surface grain size were largely independent of the nitrogen concentration and decreased with increasing bias voltage. The nanohardness and elastic modulus of each sample were measured by nanoindentation. The hardest films were produced using −150 V bias voltage and either very low (0.5-1 sccm) or very high (12-15 sccm) nitrogen flow rates.     

Synchrotron infrared radiation for electrochemical external reflection spectroscopy: a case study using ferrocyanide

Synchrotron infrared radiation has been successfully coupled through an infrared (IR) microscope to a thin-cavity external reflectance cell to study the diffusion controlled redox of a ferrocyanide solution. Excellent signal-to-noise ratios were achieved even at aperture settings close to the diffraction limit. Comparisons of noise levels as a function of aperture size demonstrate that this can be attributed to the high brilliance of synchrotron radiation relative to a conventional thermal source. Time resolved spectroscopic studies of diffusion controlled redox behavior have been measured and compared to purely electrochemical responses of the thin-cavity cell. Marked differences between the two measurements have been explained by analyzing diffusion in both the axial (linear) and radial dimensions. Whereas both terms contribute to the measured current and charge, only species that originate in the volume element above the electrode and diffuse in the direction perpendicular to the electrode surface are interrogated by IR radiation. Implications for the use of ultramicroelectrodes and synchrotron IR (SIR) to study electrochemical processes in the submillisecond time domain are discussed.     

Membrane-based parallel plate denuder for the collection and removal of soluble atmospheric gases

A continuously wetted cellulose acetate membrane-based parallel plate diffusion denuder is described. This is the first membrane-based denuder that has a small enough internal liquid holdup volume to permit reasonably rapid response time (10 --> 90% rise time of approximately 1.2 min for a transient event at a liquid flow rate of 500 microL/min) while permitting quantitative removal of common soluble atmospheric trace gases at flow rates up to 1.7 L/min. The latter attribute permits the use of the device as the first element in a particle sampling and analysis system for the quantitative removal of potentially interfering soluble trace gases. Particle losses in the denuder range from 0.9 to 2.9% over an aerodynamic diameter range of 0.38-3.48 microm, averaging 1.8%. However, only approximately 0.5% of the particles actually appears in the denuder effluent liquid. The relatively compact (300 mm H x 57 mm W x 26 mm D) wet denuder should be attractive in a number of applications. We show excellent agreement for HONO measurements with a conventional larger parallel plate wetted denuder in field measurements.     

Dielectric properties and thickness metrology of strain engineered GaN/AlN/Si (111) thin films grown by MOCVD

Maturity of silicon nanoelectronics and the high quality of 300 mm epi-Si wafers make these substrates an ideal choice for the growth of high quality III-Nitride devices. The results of our substrate engineering technique, which involves implantation of nitrogen into Si through an AlN thin film, have shown to simultaneously and significantly reduce the dislocation density and macro-cracks in epitaxially grown 2 μm GaN films. In this study, high quality strain engineered GaN films were grown by metalorganic chemical vapor deposition (MOCVD) and spectroscopic ellipsometry was used to characterize the dielectric properties, thickness, and stress of the complex structure. The uniaxial, anisotropic dielectric functions of wurtzite GaN and AlN were determined for the processes used in this study, and using this information, the thickness of each layer was determined in the completed film stack. IR spectroscopic ellipsometry (IRSE) was used as the non-destructive characterization technique to identify the IR sensitive phonon modes in AlN. The stress evolution in the films was investigated as a function of the phonon frequency shift and the broadening of the phonon modes. The results obtained by IRSE were further complemented by high resolution X-ray diffraction (HRXRD) and Raman scattering measurements.     

Physico-chemical characterization of thin films obtained by fluorination of GaAs under 5 bar of fluorine

Fluoride-GaAs(100) structures have been prepared by fluorination under 5 bar of fluorine for different temperatures and times. This paper deals with the preparation method and with the physico-chemical characterization of the samples obtained. The bulk of the fluoride layers has been investigated by Rutherford backscattering, IR absorption and Auger electron spectroscopy measurements. It has been demonstrated that for fluorine at this pressure, the best oxidation temperature is close to 200°C. The bulk composition of the films formed at the surface of GaAs substrates is GaF₃. The semiconductor-fluoride interface has been studied using the X-ray photoelectron spectroscopy technique, and arsenic has been found to be bound to fluorine.     

Combined in situ atomic force microscopy-infrared-attenuated total reflection spectroscopy

The combination of atomic force microscopy (AFM) with infrared attenuated total reflection (IR-ATR) spectroscopy for simultaneous spectroscopic evanescent field absorption and scanning probe measurements is presented. The capabilities of the combined setup are demonstrated by in situ AFM imaging of the dissolution process of urea in a cyclohexane/butanol solution with nanometer topographical resolution, while simultaneously recording the correlated bulk spectral changes by mid-infrared evanescent field absorption spectroscopy. Hence, surface modification processes such as dissolution or deposition can be simultaneously monitored by AFM imaging and IR spectroscopy in liquid environments, which has not been demonstrated to date. This combined technique will in the future enable kinetic studies on physical, chemical, and biological processes at a wide variety of surfaces providing chemical specificity via IR spectroscopy in addition to high-resolution imaging via AFM.     

Simulation of packed-bed chromatography utilizing high-resolution flow fields: comparison with models

A computer simulation of a section of the interior region of a liquid chromatographic column is performed. The detailed fluid flow profile is provided from a microscopic calculation of low Reynolds number flow through a random packed bed of nonporous spherical particles. The fluid mechanical calculations are performed on a parallel processor computer utilizing the lattice Boltzmann technique. Convection, diffusion, and retention in this flow field are calculated using a stochastic-based algorithm. This computational scheme provides for the ability to reproduce the essential dynamics of the chromatographic process from the fundamental considerations of particle geometry, particle size, flow velocity, solute diffusion coefficient, and solute retention parameters when retention is utilized. The simulation data are fit to semiempirical models. The best agreement is found for the "coupling" model of Giddings and the four-parameter Knox model. These models are verified over a wide range of particle sizes and flow velocities at both low and high velocity. The simulations appear to capture the essential dynamics of the chromatographic flow process for non-dimensional flow velocities (Péclet number) less than 500. Since the same packing geometry is utilized for different particle size studies, the interpretation of the parameter estimates from these models can be extended to the physical column model. The simulations reported here agree very well with a number of experiments reported previously.     

Mutual diffusivity in binary mixtures of n-heptane with n-hexane isomers

This paper presents a study of the influence of branching in the binary diffusion coefficients of n-heptane + n-hexane isomers, in the liquid state. The measurements have been made with the Taylor dispersion technique, at several compositions, at 283 and 298 K, for the X + n-heptane mixtures, where X= n-hexane, 3-methylpentane, 2, 3-dimethylbutane, and 2, 2-dimethylbutane. The results show a very interesting behavior of the composition dependence of the binary diffusion coefficients, presenting a maximum, for compositions about a molar fraction of n-heptane of 0.5, which increases with the increase in the degree of branching, suggesting the possibility of order-disorder effects caused by stereochemically favored packing in the liquid phase and energetically favored segment interaction in the liquid mixtures. An attempt to apply the van der Waals model to these data could not predict the experimental binary diffusion coefficients of these systems within the experimental accuracy.Paper presented at the Tenth Symposium on Thermophysical Properties, June 20–23, 1988, Gaithersburg, Maryland, U.S.A.     

Comparison between infrared and Raman spectroscopic analysis of maturing rabbit cortical bone

The molecular composition of the organic and inorganic matrices of bone undergoes alterations during maturation. The aim of this study was to compare Fourier transform infrared (FT-IR) and near-infrared (NIR) Raman microspectroscopy techniques for characterization of the composition of growing and developing bone from young to skeletally mature rabbits. Moreover, the specificity and differences of the techniques for determining bone composition were clarified. The humeri of female New Zealand White rabbits, with age range from young to skeletally mature animals (four age groups, n = 7 per group), were studied. Spectral peak areas, intensities, and ratios related to organic and inorganic matrices of bone were analyzed and compared between the age groups and between FT-IR and Raman microspectroscopic techniques. Specifically, the degree of mineralization, type-B carbonate substitution, crystallinity of hydroxyapatite (HA), mineral content, and collagen maturity were examined. Significant changes during maturation were observed in various compositional parameters with one or both techniques. Overall, the compositional parameters calculated from the Raman spectra correlated with analogous parameters calculated from the IR spectra. Collagen cross-linking (XLR), as determined through peak fitting and directly from the IR spectra, were highly correlated. The mineral/matrix ratio in the Raman spectra was evaluated with multiple different peaks representing the organic matrix. The results showed high correlation with each other. After comparison with the bone mineral density (BMD) values from micro-computed tomography (micro-CT) imaging measurements and crystal size from XRD measurements, it is suggested that Raman microspectroscopy is more sensitive than FT-IR microspectroscopy for the inorganic matrix of the bone. In the literature, similar spectroscopic parameters obtained with FT-IR and NIR Raman microspectroscopic techniques are often compared. According to the present results, however, caution is required when performing this kind of comparison.     

Investigation of optical properties of benzocyclobutene wafer bonding layer used for 3D interconnects via infrared spectroscopic ellipsometry

Benzocyclobutene (BCB) used for bonding silicon wafers to enable 3D interconnect technology is characterized using spectroscopic ellipsometry (SE). SE is a non-destructive technique that has been used to characterize the thickness and dielectric properties of BCB. The infrared (IR) absorption spectrum was used to calculate the percentage of curing of BCB on 300 mm bare and bonded wafers. The percentage of curing in BCB is a key parameter that impacts the bond strength and bond quality. This study presents the potential application of IRSE for measurements on bonded wafers to characterize the chemical information, curing percentage, bond quality and thickness of the BCB bonding layer. One of the key issues in the process development and characterization of BCB bonding for 3D interconnects of 300 mm wafers is the presence of dendrites and voids between the bonded wafers. The presence of dendrites and voids was identified by using scanning acoustic microscopy (SAM) and imaged by scanning electron microscope (SEM).     

Aerosol counterflow two-jets unit for continuous measurement of the soluble fraction of atmospheric aerosols

A new type of aerosol collector employing a liquid at laboratory temperature for continuous sampling of atmospheric particles is described. The collector operates on the principle of a Venturi scrubber. Sampled air flows at high linear velocity through two Venturi nozzles "atomizing" the liquid to form two jets of a polydisperse aerosol of fine droplets situated against each other. Counterflow jets of droplets collide, and within this process, the aerosol particles are captured into dispersed liquid. Under optimum conditions (air flow rate of 5 L/min and water flow rate of 2 mL/min), aerosol particles down to 0.3 microm in diameter are quantitatively collected in the collector into deionized water while the collection efficiency of smaller particles decreases. There is very little loss of fine aerosol within the aerosol counterflow two-jets unit (ACTJU). Coupling of the aerosol collector with an annular diffusion denuder located upstream of the collector ensures an artifact-free sampling of atmospheric aerosols. Operation of the ACTJU in combination with on-line detection devices allows in situ automated analysis of water-soluble aerosol species (e.g., NO2-, NO3-)with high time resolution (as high as 1 s). Under the optimum conditions, the limit of detection for particulate nitrite and nitrate is 28 and 77 ng/m(3), respectively. The instrument is sufficiently rugged for its application at routine monitoring of aerosol composition in the real time.     

Phase-shift fiber-loop ring-down spectroscopy

Fiber-loop ring-down spectroscopy (FLRDS) is a recently developed absorption spectroscopic technique suitable for very small liquid samples. It is based on measurements of the optical decay constant of laser intensity in a loop made of optical waveguide material. This decay constant changes as small liquid samples containing absorbing species are introduced into the loop. In this report, it is demonstrated that one can also obtain the optical decay constant using a continuous wave laser beam that is intensity modulated and then coupled into an optical fiber loop. The inherent exponential decay in the fiber loop introduces a phase shift of the light emitted from the loop with respect to the pumping beam. By measuring this phase shift, one can readily determine the concentration of the analyte introduced between the two fiber ends and a model is established to describe the relationship. It is demonstrated that this technique, dubbed phase-shift fiber-loop ring-down spectroscopy (PS-FLRDS), is well suited as an absorption detector for any flow system in which the optical absorption path is limited by the instrument architecture. By measuring the phase angle as a function of concentration of 1,1'-diethyl-4,4'-dicarbocyanine iodide in dimethyl sulfoxide, the detection limit was determined as approximately 6 microM for a 30-40-microm absorption path. A temporal resolution of approximately 100 ms was demonstrated by a rapid displacement of the solutions between the two fiber ends. Proof-of-principle use of the PS-FLRDS detection in capillary flow systems using a commercial four-way microcross established that the alignment of the fiber and the capillary can be made simple and effective, while retaining both a low detection limit and a fast response.     

High resolution scanning gate microscopy measurements on InAs/GaSb nanowire Esaki diode devices

Gated transport measurements are the backbone of electrical characterization of nanoscale electronic devices. Scanning gate microscopy （SGM） is one such gating technique that adds crucial spatial information, accessing the localized properties of semiconductor devices. Nanowires represent a central device concept due to the potential to combine very different materials. However, SGM on semiconductor nanowires has been limited to a resolution in the 50-100 nm range. Here, we present a study by SGM of newly developed III-V semiconductor nanowire InAs/GaSb heterojunction Esaki tunnel diode devices under ultra-high vacuum. Sub-5 nm resolution is demonstrated at room temperature via use of quartz resonator atomic force microscopy sensors, with the capability to resolve InAs nanowire facets, the InAs/GaSb tunnel diode transition and nanoscale defects on the device. We demonstrate that such measurements can rapidly give important insight into the device properties via use of a simplified physical model, without the requirement for extensive calculation of the electrostatics of the system. Interestingly, by precise spatial correlation of the device electrical transport properties and surface structure we show the position and existence of a very abrupt （〈10 nm） electrical transition across the InAs/GaSb junction despite the change in material composition occurring only over 30-50 nm. The direct and simultaneous link between nanostructure composition and electrical properties helps set important limits for the precision in structural control needed to achieve desired device performance.     

Simultaneous characterization of perfluoroalkyl carboxylate, sulfonate, and sulfonamide isomers by liquid chromatography-tandem mass spectrometry

A comprehensive method was developed to simultaneously separate and detect perfluorinated acid (PFA) and PFA-precursor isomers using liquid chromatography-tandem mass spectrometry (LC-MS/MS). A linear perfluorooctyl stationary phase and acidified mobile phase increased separation efficiency, relative to alkyl stationary phases, for the many perfluoroalkyl carboxylate (PFCA), perfluoroalkyl sulfonate (PFSA), and perfluorooctyl sulfonamide (PFOSA) isomers and in combination with their distinct MS/MS transitions allowed full resolution of most isomers in standards. Utilizing the absence of the "9-series" and "0-series" product ions, several perfluorooctane sulfonate (C8F17SO3-, PFOS) isomers were structurally elucidated. In human serum, only perfluorooctane sulfonamide (C8F17SO2NH2, FOSA) and PFOS consisted of significant quantities of branched isomers, whereas PFCAs were predominantly linear. Interferences that coelute with the m/z 499 --> 80 transition of PFOS on alkyl stationary phases were simultaneously separated and identified as taurodeoxycholate isomers, removal of which permitted the use of the more sensitive m/z 80 product ion and a resulting 20-fold decrease in PFOS detection limits compared to the m/z 499 --> 99 transition (0.8 pg versus 20 pg using m/z 80 and 99, respectively). Interferences in human serum which caused a 10-20-fold over-reporting of perfluorohexane sulfonate (C6F13SO3-, PFHxS) concentrations on alkyl stationary phases were also simultaneously separated from linear PFHxS and identified as endogenous steroid sulfates. PFOSA isomers, generated with human microsomes, had different rates of metabolism, suggesting that the perfluoroalkyl branching pattern may affect the biological properties of individual isomers. This fact, and for reasons of improved accuracy and sensitivity, investigators are urged to utilize more efficient separation methods capable of isomer characterization in perfluoroalkyl research.     

Theoretical modeling and experimental evaluation of a microscale molecular mass sensor

A theoretical model for a recently developed microscale molecular mass sensor (micro-MMS) is presented. The micro-MMS employs a widely applicable technique of measuring the refractive index gradient (RIG) in a microchannel created after two adjacent streams merge: a "sample stream" containing analyte(s) of interest in a host solvent and a "mobile-phase" stream containing only the host solvent. Because the flow in the microchannel is laminar, the analytes in the sample stream mix with the mobile-phase stream primarily by diffusion. The diffusion-induced RIG in the microchannel is measured by monitoring the deflection angle of a diode laser probe beam, which is orthogonal to both the direction of flow and the direction of analyte diffusion. The micro-MMS samples the RIG with probe beams at two positions along the direction of flow, and the ratio of the downstream to the upstream signal monitors the diffusion coefficient. Following calibration for a given class of compounds, the molecular mass of an analyte of interest can be determined. Along with the analyte diffusion coefficient, the theoretical model indicated three other specific parameters are important to interpret the micro-MMS output: the radius of the interrogating light probe beams, the time intervals between each of the detection positions, and the merge point relative to the detection positions. A series of experiments were conducted at different beam radii and flow rates to investigate these parameters, and the results are consistent with the model. The model shows that by using smaller beam radii and altering flow rates the molecular mass range of the micro-MMS can be, in principle, tuned from less than 10(2) g/mol to greater than 10(8) g/mol. The ratio data from the micro-MMS is also demonstrated to readily provide a "universal calibration", from which the determination of unknown diffusion coefficients can be readily obtained.