Effective absorption coefficient and effective thickness in attenuated total reflection spectroscopy.

Since the introduction of attenuated total reflection (ATR) spectroscopy for the characterization of materials, attempts have been made to relate the measured reflectivity (R) to the absorption coefficient (α) of the absorbing material of interest. The common approach is limited to the low absorption case under the assumption R∼exp(-αde), where de is an effective thickness, which is evaluated for the lossless case. In this Letter, a more detailed derivation leads to R=exp(-ßdp/2), enabling the definition of an ATR-effective absorption coefficient ß and the penetration depth dp of the electric field in the absorbing material. It is found that ß∼4πε2/λ, where ε2 is the imaginary part of the complex dielectric function of the absorbing material, and λ is the wavelength. An alternative formulation is R=exp(-αdef), where def is a generalized effective thickness for arbitrary strength of absorption which reduces to de in the low absorption limit. The experimental data for water, the biopolymer chitosan, and soda-lime glass prove the reliability of the ATR-effective absorption coefficient in the infrared range.

Structural circular birefringence and dichroism quantified by differential decomposition of spectroscopic transmission Mueller matrices from Cetonia aurata.

Transmission Mueller-matrix spectroscopic ellipsometry is applied to the cuticle of the beetle Cetonia aurata in the spectral range 300-1000 nm. The cuticle is optically reciprocal and exhibits circular Bragg filter features for green light. By using differential decomposition of the Mueller matrix, the circular and linear birefringence as well as dichroism of the beetle cuticle are quantified. A maximum value of structural optical activity of 560°/mm is found.

Sum decomposition of Mueller-matrix images and spectra of beetle cuticles.

Spectral Mueller matrices measured at multiple angles of incidence as well as Mueller matrix images are recorded on the exoskeletons (cuticles) of the scarab beetles Cetonia aurata and Chrysina argenteola. Cetonia aurata is green whereas Chrysina argenteola is gold-colored. When illuminated with natural (unpolarized) light, both species reflect left-handed and near-circularly polarized light originating from helicoidal structures in their cuticles. These structures are referred to as circular Bragg reflectors. For both species the Mueller matrices are found to be nondiagonal depolarizers. The matrices are Cloude decomposed to a sum of non-depolarizing matrices and it is found that the cuticle optical response, in a first approximation can be described as a sum of Mueller matrices from an ideal mirror and an ideal circular polarizer with relative weights determined by the eigenvalues of the covariance matrices of the measured Mueller matrices. The spectral and image decompositions are consistent with each other. A regression-based decomposition of the spectral and image Mueller matrices is also presented whereby the basic optical components are assumed to be a mirror and a circular polarizer as suggested by the Cloude decomposition. The advantage with a regression decomposition compared to a Cloude decomposition is its better stability as the matrices in the decomposition are determined a priori. The origin of the depolarizing features are discussed but from present data it is not possible to conclude whether the two major components, the mirror and the circular polarizer are laterally separated in domains in the cuticle or if the depolarization originates from the intrinsic properties of the helicoidal structure.

On liquid crystal diffractive optical elements utilizing inhomogeneous alignment.

Formation of a desired liquid crystal (LC) director distribution by the use of inhomogeneous anchoring and pre-tilt angle for electrically controlled diffractive optical elements (DOE) is studied. Such LC DOE can have high periodicity and diffraction efficiency. At the same time they are free of constructive regularities, e.g. a periodic arrangement of the electrodes or thickness deviations, which have undesired impact on diffractive characteristics of LC DOE of other types. We focus on evaluation of potential functional abilities of LC DOE with inhomogeneous alignment. The reasons causing restriction of the LC DOE diffraction efficiency and periodicity are considered. Approaches for improvement of characteristics of the LC DOE are discussed.

Fano interference in supported gold nanosandwiches with weakly coupled nanodisks.

We studied the far-field optical response of supported gold-silica-gold nanosandwiches using spectroscopic ellipsometry, reflectance and transmittance measurements. Although transmittance data clearly shows that the gold nanodisks in the sandwich structure interact very weakly, oblique reflectance spectra of s- and p-polarized light show clearly asymmetric line-shapes of the Fano type. However, all experimental results are very well described by modeling the gold nanodisks as oblate spheroids and by employing a 2 × 2 scattering matrix formulation of the Fresnel coefficients provided by an island film theory. In particular, the Fano asymmetry can be explained in terms of interference between the scattered waves from the decoupled nanodisks in the spectral range limited by their respective plasmon resonances. We also show that the reflectance and ellipsometry spectra can be described by a three-layer system with uniaxial effective dielectric functions.

Optical response of supported gold nanodisks.

It is shown that the ellipsometric spectra of short range ordered planar arrays of gold nanodisks supported on glass substrates can be described by modeling the nanostructured arrays as uniaxial homogeneous layers with dielectric functions of the Lorentz type. However, appreciable deviations from experimental data are observed in calculated spectra of irradiance measurements. A qualitative and quantitative description of all measured spectra is obtained with a uniaxial effective medium dielectric function in which the nanodisks are modeled as oblate spheroids. Dynamic depolarization factors in the long-wavelength approximation and interaction with the substrate are considered. Similar results are obtained calculating the optical spectra using the island-film theory. Nevertheless, a small in-plane anisotropy and quadrupolar coupling effects reveal a very complex optical response of the nanostructured arrays.

Optical properties of carbon nanofiber photonic crystals.

Carbon nanofibers (CNFs) are used as components of planar photonic crystals. Square and rectangular lattices and random patterns of vertically aligned CNFs were fabricated and their properties studied using ellipsometry. We show that detailed information such as symmetry directions and the band structure of these novel materials can be extracted from considerations of the polarization state in the specular beam. The refractive index of the individual nanofibers was found to be n(CNF) = 4.1.

Graded pitch profile for the helicoidal broadband reflector and left-handed circularly polarizing cuticle of the scarab beetle Chrysina chrysargyrea.

The cuticle of the beetle Chrysina chrysargyrea reflects left-handed polarized light in the broad spectral range from 340 to 1000 nm. Interference oscillations in the experimental Mueller-matrix spectroscopic ellipsometry data reveal that transparent materials comprise the cuticle. A spectral analysis of the interference oscillations makes evident that the pitch profile across the cuticle is graded. The graded pitch and effective refractive indices are determined through non-linear regression analysis of the experimental Mueller matrix by using a cuticle model based on twisted biaxial dielectric slices. Non-uniformity in cuticle thickness as well as in pitch profile near the cuticle surface account for depolarizance of the Mueller matrix. Transmission electron microscopy supports the reliability of the results.

The isoelectric point of thrombin and its behaviour compared to prothrombin at some solid surfaces.

We have shown that the isoelectric point of thrombin is high and that thrombin is a cation at the pH of blood. On the other hand, prothrombin has a low isoelectric point, being more anionic at the pH of blood. It was also found that thrombin adsorbs readily to surfaces, especially negatively charged surfaces, like behenic acid surfaces at pH 8.2. Furthermore, thrombin adsorbed onto behenic acid was active in the sense that it coagulated fibrinogen. The significance of the electric charge of the thrombin molecule in the mechanism of atherosclerosis is discussed.

Monitoring specific interaction of low molecular weight biomolecules on oxidized porous silicon using ellipsometry.

Porous silicon dioxide surfaces have been used for monitoring the specific affinity binding of low molecular weight molecules to streptavidin. Streptavidin was immobilized to the porous silicon dioxide surface by spontaneous adsorption at pH 7.4. Binding of biotin and an oligopeptide synthesized by means of combinatorial chemistry were monitored with an in situ null ellipsometer. Measurements were also done with hydroxy-azobenzene-2-carboxylic acid and DL-6-8-thioctic acid amide. The performance of porous silicon dioxide as a potential surface in biosensor applications was compared with a planar silicon dioxide surface. Porous silicon dioxide showed a 10-fold amplification of the response compared to planar silicon dioxide. It was possible to monitor the binding of biotin and the oligopeptide in the concentration range 2-40 microM. A response time as low as 30 s was obtained for the oligopeptide at 40 microM.

Non-chromogenic peptide substrates for detection of enzymes by means of capacitance changes at metal electrodes.

A new type of substrate for enzyme detection has been developed. The substrate is non-chromogenic and is used in an assay method based on electrode adsorption. The rate of change in the electric capacitance of the electrode is monitored and taken as a measure of the substrate adsorption. Substrate adsorption is in turn proportional to substrate bulk concentration and thus subject to changes by enzymes. The new substrate introduces a new concept in enzyme detection: as it is non-chromogenic it may contain appropriate amino acids on both sides of the bond subject to enzymatic cleavage.

The electrode adsorption method for determination of enzyme activity: a study of substrate requirements.

The electrode adsorption method for the determination of enzyme activity requires substrates that, besides having good kinetics constants for the enzyme, also show good adsorption/desorption kinetics to the electrode surface and adsorb in such a way that they change the double-layer capacitance of the electrode. A series of peptide substrates containing one to three aromatic groups has been synthesized. Our results show that the aromatic groups are of crucial importance for the capacitance change caused by the adsorbing/desorbing substrate. Thus, the tripeptide substrate, Bz-Phe(NO2)-Val-Arg-pNA, with three aromatic groups is superior to the other synthesized substrates containing only one or two aromatic groups. Our desorption experiments show that several factors determine the rate of capacitance increase observed when thrombin is added to a substrate solution in equilibrium with a substrate-covered electrode. The kinetic constants of the substrate determine how the substrate concentration in the solution decreases and, consequently, determine the spontaneous desorption measured as capacitance increase. Thrombin does not seem to split adsorbed substrate molecules but it adsorbs to the substrate-covered surface and in that way causes a capacitance decrease counteracting the change caused by desorption of substrate.

Penetration and loading of human serum albumin in porous silicon layers with different pore sizes and thicknesses.

Human serum albumin was adsorbed into porous silicon layers with thickness up to 3 microm and with different mean pore radius in the range 4.5-10 nm. The adsorbed amount of protein was quantified by I(125) radioactive labeling techniques and ellipsometry. The results show that albumin penetrated into the pores when the mean pore radius was larger than 5.5 nm, but could not totally occupy the available surface area when the layer thickness was larger than 1 microm. Loading of albumin both into porous layers and onto plane silicon as a function of albumin concentration was also investigated. These measurements show that loading of protein increased with protein concentration at least up to 10 mg/ml for porous silicon and up to 1 mg/ml for plane silicon. The maximum deposition into the type of porous layers used here was 28 microg/cm(2), compared to 0.36 microg/cm(2) for plane silicon.

Protein adsorption on thin films of carbon and carbon nitride monitored with in situ ellipsometry.

Thin films of amorphous carbon and amorphous, graphitic and fullerene-like carbon nitride were deposited by reactive magnetron sputtering and optically characterized with spectroscopic ellipsometry. Complementary studies using scanning electron microscopy and atomic force microscopy were performed. The films were exposed to human serum albumin (HSA) and the adsorption was monitored in situ using dynamic ellipsometry. From the ellipsometric data the adsorbed amount of proteins was quantified in terms of surface mass density using de Feijter's model. The results indicate larger adsorption of proteins onto the amorphous films compared to the films with a more textured structure. Complementary studies with 125I-labeled HSA showed an apparent protein adsorption up to six times larger compared to the ellipsometry measurement. In addition, the four types of films were incubated in blood plasma followed by exposure to anti-fibrinogen, anti-HMWK or anti-C3c, revealing the materials' response to complement and contact activation. The amorphous and graphitic carbon nitride exhibit rather high immune activity compared to a titanium reference, whereas the amorphous carbon and the fullerene-like CNx show less immune complement deposition. Compared to the reference, all films exhibit indications of a stronger ability to initiate the intrinsic pathway of coagulation. Finally, the surfaces' bone-bonding ability was investigated by examination of their ability to form calcium phosphate crystals in a simulated body fluid, with a-CNx depositing most calcium phosphate after 21 days of incubation.

Formation and cross-linking of fibrinogen layers monitored with in situ spectroscopic ellipsometry.

Thick matrices of fibrinogen with incorporation of a matrix metalloproteinase inhibitor were covalently bonded on functionalized silicon surfaces using an ethyl-3-dimethyl-aminopropyl-carbodiimide and N-hydroxy-succinimide affinity ligand coupling chemistry. The growth of the structure was followed in situ using dynamic ellipsometry and characterized at steady-state with spectroscopic ellipsometry. The growth was compared with earlier work on ex situ growth of fibrinogen layers studied by single wavelength ellipsometry. It is found that in situ growth and ex situ growth yield different structural properties of the formed protein matrix. Fibrinogen matrices with thicknesses up to 58 nm and surface mass densities of 1.6 microg/cm2 have been produced.