Performance of a highly sensitive optical waveguide Mach-Zehnder interferometer immunosensor

We describe a highly sensitive sensor which uses the evanescent field of a reusable planar optical waveguide as the sensing element. The waveguide used is optimized to obtain a steep dependence of the propagation velocity on the refractive-index profile near the surface. The adsorption of a layer of proteins thus results in a phase change, which is measured interferometrically using a Mach-Zehnder interferometer set-up. The stability of the interferometer is such that phase changes = (1 × 10-2)2pi per hour can be measured. Immunoreactions have been monitored down to concentrations of 5 × 10-11 M of a 40 kDa protein

Development of an optical waveguide interferometric immunosensor

A Mach-Zehnder interferometer is presented, which uses the evanescent field of a planar optical waveguide as the sensing element. Changes in the refractive index profile occurring in the immediate vicinity of the waveguide surface cause the propagation velocity of the light passing through it to change. This is measured interferometrically. Adsorption processes of antibodies onto the waveguide surface as well as immunoreactions have been monitored. Concentrations of 2.5 × 10¿8 M (1 × 10-3 mg/ml) of the antigen HCG (molecular weight = 39 600) have been measured so far

Refractive index and layer thickness of an adsorbing protein as reporters of monolayer formation

A method is presented for a separate real-time determination of refractive index and layer thickness of an adsorbing thin layer. The changing angular deflections of TE and TM modes in a dedicated planar waveguide structure are measured. A resolution of 0.01 in the refractive index and 0.5 nm in the average thickness is obtained. The method is illustrated with experimental results on the binding of an antibody to the substrate, both in a physisorption and in an immunoreaction. In the latter, results are consistent with an end-on binding of the antibody to the surface

Temperature dependence of Raman vibrational bandwidths in poly(rA) and rAMP

Isotropic and anisotropic spontaneous Raman spectra were obtained from solutions of poly(ra) and rAMP in buffer. The temperature dependence of these spectra was measured to elucidate the influence of macromolecular dynamics and solvent dynamics on the bandwidths of base vibrations in the single stranded polynucleotide poly(rA). The temperature dependence of a bandwidth depends upon the particular vibration under study. The bands can for the larger part be described by Lorentz functions. When fitted by Voigt functions, maximally 10% of each bandprofile of the adenine base vibrations can be attributed to a Gaussian component. The second moment has been determined from the spectra for the 725 cm¿1 band. From the second moment and the bandwidth, we were able to deduce that the vibrational oscillator is in the fast modulation limit. The determined timescale (perturbation correlation time 0.13 ps) eliminate perturbations connected to long range diffusion like concentration fluctuations (timescale in the order of 10 ps). The spectra were analyzed by an extensive curve fitting procedure providing accurate bandparameters (position, width and integrated intensity). The 725 cm¿1 band of adenine has a bandwidth which is dependent upon the degree of polymerization. In RAMP it is 17.6 cm¿1, in stacked (i.e. low temperature 5°C) poly(rA) it is 11.5 cm¿1. The bandwidth of the adenine vibration at 1336 cm¿1 cm¿1 has a temperature dependence which is similar to the intensity changes of the Raman and the absorption hypochromic effect as a function of temperature. The melting transition can therefore be followed by the changes in bandwidth of suitable vibrations

An improved optical method for surface plasmon resonance experiments

In this paper an inexpensive optical device is described, which is capable of measuring the optical reflectance at different angles, while keeping the laser spot stationary at one point of the surface. This is accomplished by applying cylindrical optics. Its use is demonstrated in a surface plasmon resonance (SPR) sensor. A coil-operated vibrating mirror is used to obtain an angle scan of about 4 degrees. The angle of minimum reflectance can be detected with an accuracy of approximately 2 × 10−3 degrees. Despite the use of simple optical components, disturbance of laser beam parallelism is no more than 0.02 degrees. Displacement of the laser spot at the surface during the angle scan is kept within 0.2 mm. The device eliminates disturbances due to surface irregularities in measurements.\u

Accretion disks in Algols: progenitors and evolution

There are only a few Algols with measured accretion disk parameters. These measurements provide additional constraints for tracing the origin of individual systems, narrowing down the initial parameter space. We investigate the origin and evolution of 6 Algol systems with accretion disks to find the initial parameters and evolutionary constraints for them. With a modified binary evolution code, series of close binary evolution are calculated to obtain the best match for observed individual systems. Initial parameters for 6 Algol systems with accretion disks were determined matching both the present system parameters and the observed disk characteristics. When RLOF starts during core hydrogen burning of the donor, the disk lifetime was found to be short. The disk luminosity is comparable to the luminosity of the gainer during a large fraction of the disk lifetime.Comment: 8 pages, 6 figures , accepted by A&

A Raman spectroscopic study of the interaction between nucleotides and the DNA binding protein gp32 of bacteriophage T4

Raman spectra of the bacteriophage T4 denaturing protein gp32, its complex with the polynucleotides poly(rA), poly(dA), poly(dT), poly(rU), and poly(rC), and with the oligonucleotides (dA)8 and (dA)2, were recorded and interpreted. According to an analysis of the gp32 spectra with the reference intensity profiles of Alix and co-workers [M. Berjot, L. Marx, and A.J.P. Alix (1985) J. Ramanspectrosc., submitted; A.J.P. Alix, M. Berjot, and J. Marx (1985) in Spectroscopy of Biological Molecules, A. J. P. Alix, L. Bernard, and M. Manfait, Eds., pp. 149-154], 1 gp32 contains ≈ 45% helix, ≈ 40% β-sheet, and 15% undefined structure. Aggregation of gp32 at concentrations higher than 40 mg/mL leads to a coordination of the phenolic OH groups of 4-6 tyrosines and of all the sulfhydryl (SH) groups present in the protein with the COO- groups of protein. The latter coordination persists even at concentrations as low as 1 mg/mL. In polynucleotide-protein complexes the nucleotide shields the 4-6 tyrosine residues from coordination by the COO- groups even at high protein concentration. The presence of the nucleotide causes no shielding of the SH groups. With Raman difference spectroscopy it is shown that binding of the protein to a single-stranded nucleotide involves both tyrosine and trytophan residues. A change in the secondary structure of the protein upon binding is observed. In the complex, gp32 contains more -sheet structure than when uncomplexed. A comparison of the spectra of complexed poly(rA) and poly(dA) with the spectra of their solution conformations at 15°C reveals that in both polynucleotides the phosphodiester vibration changes upon complex formation in the same way as upon a transition from a regular to a more disordered conformation. Distortion of the phosphate-sugar-base conformation occurs upon complex formation, so that the spectra of poly(rA) and poly(dA) are more alike in the complex than they are in the free polynucleotides. The decrease in intensity of the Raman bands at 1304 cm-1 in poly(rA), at 1230 cm-1 in poly(rU), and at 1240 and 1378 cm-1 of poly(dT) may be indicative of increased stacking interactions in the complex. No influence of the nucleotide chain length upon the Raman spectrum of gp322 in the complex was detected. Both the nucleotide lines and the protein lines in the spectrum of a complex are identical in poly(dA) and (dA)8

The effect of intermediate mass close binaries on the chemical evolution of Globular Clusters

Context. The chemical processes during the Asymptotic Giant Branch (AGB) evolution of intermediate mass single stars predict most of the observations of the different populations in globular clusters although some important issues still need to be further clarified. In particular, to reproduce the observed anticorrelations of Na-O and Al-Mg, chemically enriched gas lost during the AGB phase of intermediate mass single stars must be mixed with matter with a pristine chemical composition. The source of this matter is still a matter of debate. Furthermore, observations reveal that a significant fraction of the intermediate mass and massive stars are born as components of close binaries. Aims. We will investigate the effects of binaries on the chemical evolution of Globular Clusters and on the origin of matter with a pristine chemical composition that is needed for the single star AGB scenario to work Methods. We use a population synthesis code that accounts for binary physics in order to estimate the amount and the composition of the matter returned to the interstellar medium of a population of binaries. Results. We demonstrate in the present paper that the mass lost by a significant population of intermediate mass close binaries in combination with the single star AGB pollution scenario may help to explain the chemical properties of the different populations of stars in Globular Clusters.Comment: 11 pages, 4 figures. Paper accepted for Astron. & Astrophy