Biosensing using porous silicon double-layer interferometers: reflective interferometric Fourier transform spectroscopy

A simple, chip-based implementation of a double-beam interferometer that can separate biomolecules based on size and that can compensate for changes in matrix composition is introduced. The interferometric biosensor uses a double-layer of porous Si comprised of a top layer with large pores and a bottom layer with smaller pores. The structure is shown to provide an on-chip reference channel analogous to a double-beam spectrometer, but where the reference and sample compartments are stacked one on top of the other. The reflectivity spectrum of this structure displays a complicated interference pattern whose individual components can be resolved by fitting of the reflectivity data to a simple interference model or by fast Fourier transform (FFT). Shifts of the FFT peaks indicate biomolecule penetration into the different layers. The small molecule, sucrose, penetrates into both porous Si layers, whereas the large protein, bovine serum albumin (BSA), only enters the large pores. BSA can be detected even in a large (100-fold by mass) excess of sucrose from the FFT spectrum. Detection can be accomplished either by computing the weighted difference in the frequencies of two peaks or by computing the ratio of the intensities of two peaks in the FFT spectrum.     

Fourier transform raman spectroscopy using a bench-top fourier transform infrared spectrometer

The use of a bench top FTIR spectrometer for near infrared Fourier transform Raman spectroscopy is demonstrated. The use of near infrared excitation results in fluorescence free Raman spectra allowing previously difficult samples to be measured.     

Characterization of phospholipid bilayer formation on a thin film of porous SiO2 by reflective interferometric Fourier transform spectroscopy (RIFTS)

Classical methods for characterizing supported artificial phospholipid bilayers include imaging techniques such as atomic force microscopy and fluorescence microscopy. The use in the past decade of surface-sensitive methods such as surface plasmon resonance and ellipsometry, and acoustic sensors such as the quartz crystal microbalance, coupled to the imaging methods, have expanded our understanding of the formation mechanisms of phospholipid bilayers. In the present work, reflective interferometric Fourier transform spectrocopy (RIFTS) is employed to monitor the formation of a planar phospholipid bilayer on an oxidized mesoporous Si (pSiO(2)) thin film. The pSiO(2) substrates are prepared as thin films (3 μm thick) with pore dimensions of a few nanometers in diameter by the electrochemical etching of crystalline silicon, and they are passivated with a thin thermal oxide layer. A thin film of mica is used as a control. Interferometric optical measurements are used to quantify the behavior of the phospholipids at the internal (pores) and external surfaces of the substrates. The optical measurements indicate that vesicles initially adsorb to the pSiO(2) surface as a monolayer, followed by vesicle fusion and conversion to a surface-adsorbed lipid bilayer. The timescale of the process is consistent with prior measurements of vesicle fusion onto mica surfaces. Reflectance spectra calculated using a simple double-layer Fabry-Perot interference model verify the experimental results. The method provides a simple, real-time, nondestructive approach to characterizing the growth and evolution of lipid vesicle layers on the surface of an optical thin film.     

Far infrared fourier transform spectroscopy of semiconductors

Fourier transform spectroscopy (FTS) is one of the most important tools in the study of shallow level donors and acceptors in semiconductors. When combined with a two-step photothermal ionization process detected photoconductively, FTS allows measurement of optical transitions of donor-bound electrons (and acceptor-bound holes) in ultra-pure germanium samples with impurity concentrations <10⁹ cm^–3 (i.e. one electrically active impurity in 4×10¹³ host atoms). The experimental high resolution study of the hydrogen-like excited state series of shallow levels has yielded as many as 19 lines of width as small as 10eV for some centers. These results have stimulated theoretical work which has led to the unambiguous assignment of quantum states to many bound excited states. Extensive studies of ultra-pure Ge crystals grown under different well-controlled conditions have led to the discovery of a large number of novel shallow impurity complexes. Study of the multiplicities and symmetries of the associated electronic states has led to a detailed understanding of the unusual static and dynamic structures of these novel centers. The chemical composition has been deduced from correlations between the concentration of a particular center and the materials involved in crystal gowth. Isotopic substitution of hydrogen with deuterium has led to the unambiguous proof of the presence of hydrogen in several of the novel centers. In addition to the high resolution spectra of shallow electronic levels, vibrational spectra of bond-centered interstitial oxygen in ultra-pure Ge are noteworthy for their extraordinarily sharp lines.     

Development of an enrofloxacin immunosensor based on label-free electrochemical impedance spectroscopy

Enrofloxacin is the most widespread antibiotic in the fluoroquinolone family. As such, the development of a rapid and sensitive method for the determination of trace amounts of enrofloxacin is an important issue in the health field. The interaction of the enrofloxacin antigen to a specific antibody (Ab) immobilized on an 11-mercapto-undecanoic acid-coated gold electrode was quantified by electrochemical impedance spectroscopy. Two equivalent circuits were separately used to interpret the obtained impedance spectra. These circuits included one resistor in series with one parallel circuit comprised of a resistor and a capacitor (1R//C), and one resistor in series with two parallel RC circuits (2R//C). The results indicate that the antigen-antibody reaction analyzed using the 1R//C circuit provided a more sensitive resistance increment against the enrofloxacin concentration than that of the 2R//C circuit. However, the 2R//C circuit provided a better fitting for impedance spectra, and therefore supplies more detailed results of the enrofloxacin-antibody interaction, causing the increase of electron transfer resistance selectively to the modified layer, and not the electrical double layer. The antibody-modified electrode allowed for analysis of the dynamic linear range of 1-1000 ng/ml enrofloxacin with a detection limit of 1 ng/ml. The reagentless and label-free impedimetric immunosensors provide a simple and sensitive detection method for the specific determination of enrofloxacin.     

The destruction-free analysis of polymers by fourier transform infrared photoacoustic and fourier transform Raman spectroscopy: A comparison

With the introduction of rapid–scanning Fourier transform infrared (FTIR) and recently Raman (FT–Raman) spectroscopy, vibrational spectroscopy has been launched into a new era of applications in polymer chemistry and physics. Thus, the increase in sensitivity provided by multiple scanning has led to the breakthrough of new, destruction–free sampling techniques, such as photoacoustic and Raman spectroscopy. This paper provides a comparison between data produced by FTIR photoacoustic and FT–Raman analysis of a range of polymers, and structural information available from both techniques is discussed.     

Near infrared fourier transform Raman spectroscopy of human artery

We report the first near IR FT-Raman spectroscopy of normal diseased human artery. In normal human aorta, two bands at 1669 cm⁻¹ and 1452 cm⁻¹ dominate the spectrum and can be assigned to protein amide I and C-H in-plane bending vibrations, respectively. Weaker bands are also observed between 1250 and 1350 cm⁻¹. Non-calcified atherosclerotic lesions with a large amount of necrotic debris below the tissue surface show a relative increase in the intensity of the 1452 cm⁻¹ band. In atherosclerotic aortas which contain calcified deposits several hundred microns below the tissue surface, a strong 961 cm⁻¹ band is observed due to the symmetric stretch of phosphate groups in the calcified salts. The results show that this method provides the capability to probe biological substituents several hundred microns below the tissue surface.     

Fabrication of a label-free electrochemical immunosensor of low-density lipoprotein

The silver chloride@polyaniline (PANI) core-shell nanocomposites (AgCl@PANI) combined with Au nanoparticles (AuNPs) were used to prepare the AuNPs-AgCl@PANI hybrid material. A novel sensitive label-free low-density lipoprotein (LDL) electrochemical biosensor was fabricated by adsorption of antibody to apolipoprotein B-100 (aopB-100) on an AuNPs-AgCl@PANI-modified glassy carbon (GC) electrode. The hybrid material could provide surface for high antibody loading due to its large surface-to-volume ratio. Since each LDL has an apoB-100 on its phospholipids coat, they could be bonded to the electrode surface through the specific antibody-antigen reaction. Electrochemical impedance spectroscopy (EIS) was used to characterize the recognition of LDL. The negative charges carried by LDL phospholipids coat would block the electron transfer of the [Fe(CN)6]3-/4- redox couple severely. In addition, the conductivity of LDL is very poor, so small amounts of LDL on the electrode could result in great change in the electron-transfer resistance (Ret). The biosensor exhibited a highly sensitive response to LDL with a detection limit of 0.34 pg/mL, and some factors that would affect the performance of the biosensor were studied, such as incubation time and temperature.     

Biological applications of fourier transform infrared spectroscopy: a study of phase transitions in biomembranes

The thermal response of aqueous lipid membranes has been investigated by Fourier transform infrared spectroscopy. Changes in infrared spectral parameters are applied to the analysis of the strutural changes which occur within the lipid bilayer as the temperature is varied. Such studies provide the basis for the interpretation of phase transitions in complex biomembranes.     

Depth-profiling of polymer laminates using fourier transform infrared (ATR) spectroscopy: The barrier film technique

We report FTIR-ATR spectra of a polymer laminate (PMMA/PVOH) at different base layer thicknesses and different angles of incidence on a ZnSe substrate. By varying this ‘barrier’ layer (of PMMA) we have demonstrated that different effective penetration depths into the PVOH are achieved, in very good agreement with the calculated electric fields as a function of distance away from the substrate surface. Such work forms the basis for depth profiling measurements in order to detect interfacial interactions between the polymer molecules. © 1994 John Wiley & Sons, Inc.     

Characterization by fourier transform infrared spectroscopy (FT-IR) and 2D IR correlation spectroscopy of PAMAM dendrimer

FT-IR and 2D correlation spectroscopy were employed to study the microstructural changes occurring during phase transitions of a liquid crystal poly(amidoamine) codendrimer (PAMAM (L1)16(L2)16) generation 3, functionalized on the terminal groups by one-chain promesogenic calamitic units (4-(4'-decyloxybenzoyloxy)salicylaldehyde (L1)) and two-chain promesogenic calamitic units (4-(3',4'-didecyloxybenzoyloxy)salicylaldehyde (L2)). Spectral modifications associated with molecular conformation rearrangements allowing for molecular shape change on going from a liquid-crystalline organization to another were found. The transition temperatures were calculated, and they are in good agreement with the DSC data. Spectral analysis gives evidence of the LC phase transitions and to an additional transition associated with the existence of conformers. Various types of hydrogen bonding have been established.     

Catalytic isomerization of quadricyclane using fourier transform near-infrared absorption spectroscopy: diffusion, conversion, and temperature effect

By using Fourier transform near-infrared (NIR) absorption spectroscopy, the kinetic behaviors of quadricyclane isomerization, as catalyzed by anhydrous CuSO(4) in chloroform mixture with and without agitation, are presented. Given the acquired NIR spectra, the concentration decay of quadricyclane with the reaction time is determined with the aid of partial least-squares analysis. When the mixture is not agitated, the diffusion coefficients in chloroform are evaluated to be (3.8 +/- 0.1) x 10(-5) cm(2) s(-1) at 27 degrees C and (4.4 +/- 0.1) x 10(-5) cm(2) s(-1) at 39 degrees C. In the size-dependent experiments of the catalyst, the one-site and two-site coordinated conversion rate constants are further determined to be (8.5 +/- 5.9) x 10(-6) s(-1) A(-1) and (2.2 +/- 0.8) x 10(-8) s(-1) A(-2), respectively, at 27 degrees C and (1.3 +/- 0.8) x 10(-5) s(-1) A(-1) and (1.92 +/- 0.01) x 10(-6) s(-1) A(-2), respectively, at 39 degrees C. A denotes the total catalyst surface area per unit effective volume of solvent. Accordingly, the activation energies for one-site and two-site coordination are evaluated to be 24.8 and 286.2 kJ mol(-1), respectively. The reaction is dominated by one-site coordination (1:1 complex) between the reactant and the catalyst. Unless temperature increases, the two-site coordinated reaction may be ignored. In contrast, when analogous experiments are performed in the stirred solution, the diffusion factor is ignored but the conversion rate constants rise due to the increase of collision frequency. For instance, the one-site and two-site coordinated rate constants are increased to (1.7 +/- 1.4) x 10(-5) s(-1) A(-1) and (1.27 +/- 0.06) x 10(-5) s(-1) A(-2) at 39 degrees C. The two-site coordinated reaction rate is enhanced by a factor of 10. Thus, isomerization may proceed via both 1:1 and 1:2 coordination between the reactant and the catalyst. The Arrhenius plot yields the corresponding activation energies to be 24 +/- 3 and 275 +/- 3 kJ mol(-1). The activation energies remain constant, no matter whether the solution is agitated or not.     

Two-dimensional fourier transform spectroscopy of adenine and uracil using shaped ultrafast laser pulses in the deep UV

We compare two-dimensional (2D) ultrafast Fourier transform spectroscopy measurements in the deep UV (262 nm) for adenine and uracil in solution. Both molecules show excited-state absorption on short time scales and ground-state bleaching extending for over 1 ps. While the 2D spectrum for uracil shows changes in the center of gravity during the first few hundred femtoseconds, the center of gravity of the 2D spectrum for adenine does not show similar changes. We discuss our results in light of ab initio electronic structure calculations.     

Sandwich-type amperometric immunosensor for human immunoglobulin G using antibody-adsorbed Au/SiO2 nanoparticles

A novel sandwich-type electrochemical immunosensor for human immunoglobulin G (hIgG) was developed using Au/SiO₂ nanoparticles (NPs) with adsorbed horseradish peroxidase-anti-hIgG as the secondary antibody layer. The signal readout is based on the amperometric response to the catalytic reduction of hydrogen peroxide at an AuNPs-polythionine modified glassy carbon electrode. Under optimized conditions, the linear range is from 0.1 to 200 ng·mL^?1, with a detection limit of 0.035 ng·mL^?1 (at an S/N of 3). The immunosensor exhibited a performance that is better than that based on Au/SiO₂NPs-excluded secondary antibody.     

Macroporous ordered titanium dioxide (TiO2) inverse opal as a new label-free immunosensor

Photonic crystal sensing materials have been validated that they are very sensitive to refractive index changes. Herein, three-dimensionally ordered macroporous (3DOM) (>50 nm) TiO₂ inverse opal film has been fabricated by the self-assembly technique. Based on the TiO₂ inverse opal film, the optical spectrometer was established for label-free immunosensor. The sensing performance of the 3DOM TiO₂ was investigated using human IgG/goat anti-human IgG couple, which showed that the sensitivity of 3DOM TiO₂ inverse opal film could reach to 1 μg mL⁻¹ (equivalent to 1.5 pg mm⁻²) of protein concentration detection limit. The 3DOM TiO₂ inverse opal has a large internal surface area, low fluorescence background and unique optical properties. These characteristics indicated the feasibility of 3DOM TiO₂ inverse opal in label-free immunoassay.     

涂石蜡大米傅立叶变换红外吸收光谱识别分析

针对11种未涂石蜡大米和18个涂不同量石蜡的大米样品,以石油醚提取的油脂为试样,采用傅立叶变换红外光谱仪,扫描样品的傅立叶变换红外吸收光谱,并对光谱进行预处理,提取红外特征信息,将2855与1746、1462与1163 cm-1处特征峰的面积比值为坐标,采用Origín 6.0软件作识别分类图.结果表明:特征峰的面积比值与所涂石蜡量成线性变化,大米油脂的特征峰面积比值在一定的区域分布,涂以0.05%以上石蜡的大米,其油脂特征峰面积比值与未涂石蜡米油脂的值有一定区别.     

Melamine–formaldehyde resins: Constitutional characterization by fourier transform 13C-NMR spectroscopy

The random chemical structures of melamine–formaldehyde resins, including methylated melamine–formaldehyde resins and urea–melamine formaldehyde resins, were investigated by ¹³C-NMR spectroscopy (Fourier transform). All the combined formaldehydes, methylol and methyl ether groups, methylene structures, and dimethylene ether structures were assigned. A ¹³C chemical shift of methylene carbon occurred by substitution of other constituents of the methylene group for a proton of the adjacent monosubstituted nitrogen atom, as shown in a ¹³C-NMR spectrum of urea–formaldehyde resins. It was found that the chemical shift of each corresponding carbon of both melamine resins and urea–melamine resins was almost superimposed with that of urea resins.     

An interlaboratory comparison of sample presentation methods for the analysis of aqueous solutions using fourier transform infrared spectroscopy

Fourier transform infrared (FTIR) spectroscopy is becoming much more widely used as an analytical technique. It appears to be particularly useful for quantitative analysis, especially of aqueous solutions. However, there is a considerable variation in the cost and quality of instrumentation as well as a large number of accessories for sample presentation employing attenuated total reflectance (ATR). The objectives of the work reported in this paper were to investigate the effect of the choice of instrumentation on the quality of quantitative analyses as well as the effect of variable acquisition parameters. A series of experiments was designed in which samples were distributed to a number of workers using a range of FTIR instruments from different manufacturers and of differing quality. The workers were allowed to use the accessory of their own choice and to carry out a quantitative analysis of glucose solutions using their normal operating parameters. In following experiments the effects of instrumental and purge were considered and quantitative analysis was performed using a set of prescribed acquisition conditions. The results showed that there were significant differences in the quality of the spectra produced on the different combinations. However, although each worker produced quite different linear calibrations (in terms of slope and intercept), they all predicted the concentration of an unknown with a similar accuracy. However, later experiments showed that the low-cost bench-top instruments were not always so successful when the peaks used in the analysis were affected by residual water vapour interferences. Instrumental drift was noticeably worse for the low-cost instruments but this was an effect that could easily be eliminated by suitable baseline correction. Received: 12 December 1994/Accepted: 13 February 1995 Correspondence to: R.H. Wilson