Optics: measuring and testing – For optical fiber or waveguide inspection
Patent
1991-05-10
1994-05-17
Evans, F. L.
Optics: measuring and testing
For optical fiber or waveguide inspection
2504581, 356318, 356446, 356246, 356445, G01N 2117, G01N 3353
Patent
active
053132645
DESCRIPTION:
BRIEF SUMMARY
The present invention relates to an optical multi-analyte biosensor system employing the principle of internal reflection of polarized light for use in biological, biochemical and chemical analysis and in particular for detecting a specific molecule, for example antigens. The detection method used in the biosensor system may be based on the evanescent wave phenomenon at total internal reflection, such as surface plasmon resonance (SPR), critical angle refractometry, total internal reflection fluorescence (TIRF), total internal reflection phosphorescence, total internal reflection light scattering, and evanescent wave ellipsometry. Furthermore, the detection method may be based on Brewster angle reflectometry.
The main advantage of the internal reflection based techniques is that the sensitivity region for the specific substance is restricted to the extension length of an evanescent wave, i.e. the depth of an electromagnetic wave penetrating into the liquid medium from the sensing surface side. Consequently, there will be a minimum of influence on the response connected to specifically bound analyte molecules from non-bound sample molecules. Moreover, the penetration depth of the evanescent wave for a totally reflected ray of light depends on the angle of incidence for the ray. For a comprehensive treatment of the concept of internal reflection one is referred to Mirabella and Harrick, Internal Reflection Spectroscopy, Harrick Scientific Corporation, N.Y. 1985.
The optical response induced by either the primary evanescent wave, or by a secondary evanescent wave excited in turn by said primary evanescent wave, may be measured as changes in the reflectance or in the state of polarization of the incident light wave upon reflection, or as the fluorescence or phosphorescence or light scatter of radiation, as a result of a specific substance interaction with a sensing layer at the sensing surface.
The optical response related to the specific substance may be measured as the reflected intensity as a function of angle of incidence of p-polarized light, being applicable for surface plasmon resonance, Brewster angle reflectometry, and critical-angle refractometry, in that the angle of minimum reflectance is determined and related to a refractive index and surface concentration of the bound substance at the sensing surface.
With regard a detection using surface plasmon resonance, this will be treated in more detail hereinafter.
Internal multiple-angle Brewster angle reflectometry, based on rotating optical means and a rotating prism for the variation of incident angle, has been shown to provide information to characterize the adsorption of proteins on a silica prism/solution interface; see Schaaf, P. et al. Langmuir, 3, 1131-1135 (1987).
Critical-angle refractometry has so far been used mainly to measure the concentration or density of solutions in process streams.
In a detection employing evanescent wave ellipsometry, the optical response related to the specific substance is measured as changes in the state of polarization of elliptically polarized light upon reflection, in that this state of polarization is related to a refractive index, thickness, and surface concentration of a bound sample at the sensing surface. Multiple-angle evanescent wave ellipsometry, in the form of using rotating optical means and a rotating prism for the variation of incident angle, and a phase-modulated ellipsometer, has been used for studying the polymer (polystyrene) concentration profile near a prism/liquid interface; see Kim, M. W., Macromolecules, 22, (1989) 2682-2685. Furthermore, total internal reflection ellipsometry in the form of stationary optical means at a single angle of incidence has been suggested for quantification of immunological reactions; see EP-A1-0 067 921 (1981), and EP-A1-0 278 577 (1988).
By a detection employing evanescent wave excitation fluorescence through an angle of incidence dependent TIRF, the intensity and wavelength of the radiation emitted from the either natively fluorescent or fluorescence labeled sa
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Ivarsson Bengt
Sjodin Hakan
Sjolander Stefan
St.ang.hlberg Ralph
Ulf Jonsson
Evans F. L.
Pharmacia Biosensor AB
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