Surface Plasmon Microscopy of Two Crystalline Domains in a Lipid Monolayer

Surface plasmon microscopy is applied to monolayers of dimyristoylphosphatidylethanolamine,dipcast at high lateral pressure (35 mN/m) on a solid substrate. The vertical resolution of better than 1 nm and an in-plane resolution of - 10 um allow for the detection of two separate solid domains. Assuming a fixed value for the lipid refractive index, the tilt angles of the lipid aliphatic chains were determined to be 0° and -35°, respectively

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

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

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

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

A new approach to immunoFET operation

A new method is presented for the detection of an immunological reaction taking place in a membrane, which covers the gate area of an ISFET. By stepwise changing the electrolyte concentration of the sample solution, a transient diffusion of ions through the membrane-protein layer occurs, resulting in a transient membrane potential, which is measured by the ISFET. The diffusion rate is determined by the immobile charge density in the amphoteric protein layer, which changes upon formation of antibody—antigen complexes. No membrane potential is induced at zero fixed charge density as occurs at a protein characteristic pH. Isoeletric points of embedded proteins can be determined by detecting the zero potential response.\ud \ud Up to now, the authors have studied the membrane adsorption of lysozyme, human serum albumin (HSA) and the immune reaction of HSA with the antibody anti-human serum albumin (αHSA). The influence of protein parameters on the amplitude of the transient can be described with an empirical equation. Assuming Langmuir behaviour, the protein concentration in the solution can well be correlated with the concentration in the membrane.\ud \ud This new detection method is unique concerning direct measurements of charge densities and isoelectric points of amphoteric macromolecules adsorbed in the membrane. The simple procedure of one incubation stage followed by one detection stage, without separate washing and labelling techniques, gives direct information about specific charge properties of the macromolecules to be studied

New detection method for atrazine pesticides with the optical waveguide Mach-Zehnder immunosensor

Concentrations of analytes can be determined within a few minutes using on-line analysis of the immunobinding kinetics in a solid phase immunoassay. This approach has been applied to the detection of atrazine. Atrazine is detected, at concentrations around the European Community limit (0.1 ¿g/l) by a competitive assay. To this end, the two channels of a Mach-Zehnder waveguide sensor are used simultaneously in a difference measurement. The advantage of this way of measuring is discussed with the atrazine measurements