Integrated optic waveguide immunosensor
First Claim
1. An apparatus for performing a specific binding assay, the apparatus comprising:
- a composite waveguide comprising;
a substrate comprising a first optical material of refractive index n1 and having a first planar surface and an opposite second surface separated by a thickness and a surrounding edge, and a waveguide film comprising a second optical material having a refractive index n2 which is greater than n1, said waveguide film disposed on said first planar surface of said substrate;
capture molecules, associated with said waveguide film, for interacting selectively with at least one type of selected analyte molecule;
a light source operably disposed to direct a light beam into said composite waveguide for propagation by total internal reflection therein;
a light detection device positioned in a cone of collection angles, said cone of collection angles having an axis oriented substantially orthogonal to a plane of said waveguide film.
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Abstract
A composite waveguide for evanescent sensing in fluorescent binding assays comprising a substrate layer having one or more thin film waveguide channels deposited thereon. Binding molecules having the property of binding with specificity to an analyte are immobilized on the surface of the thin film channels. In preferred embodiments, the composite waveguide includes integral light input coupling means adapted to the thin film channels. Light coupling means may include a grating etched into the substrate prior to deposition of the thin film, or a waveguide coupler affixed to the upper surface of the thin film. The waveguide coupler has an input waveguide of high refractive index which receives the laser light through one end, propagating it by total internal reflection. Propagated light is coupled evanescently into the thin film across a spacer layer of precise thickness with a lower index of refraction than the input waveguide or the thin-film waveguide.
119 Citations
63 Claims
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1. An apparatus for performing a specific binding assay, the apparatus comprising:
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a composite waveguide comprising;
a substrate comprising a first optical material of refractive index n1 and having a first planar surface and an opposite second surface separated by a thickness and a surrounding edge, and a waveguide film comprising a second optical material having a refractive index n2 which is greater than n1, said waveguide film disposed on said first planar surface of said substrate;
capture molecules, associated with said waveguide film, for interacting selectively with at least one type of selected analyte molecule;
a light source operably disposed to direct a light beam into said composite waveguide for propagation by total internal reflection therein;
a light detection device positioned in a cone of collection angles, said cone of collection angles having an axis oriented substantially orthogonal to a plane of said waveguide film. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51)
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26. An apparatus for performing specific binding assays, the apparatus comprising:
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a light source providing light of a first wavelength;
a composite waveguide comprising a substrate having a first planar surface and an opposite second surface, said substrate comprising a first optical material of refractive index n1, and a waveguide film disposed on said first planar surface, said waveguide film comprising a second optical material of refractive index n2 and including a first surface with a plurality of capture molecules secured thereto, each capture molecule of said plurality of capture molecules having a binding site which selectively binds a corresponding, selected analyte, said waveguide film also including a second surface adjacent to said first planar surface of said substrate;
a light detector positioned to detect light of a second wavelength emitted through at least said waveguide film, said first and second wavelengths differing from one another.
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52. A composite waveguide comprising:
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a substrate comprising a first optical material of refractive index n1 and having a first planar surface and an opposite second surface separated by a thickness and a surrounding edge;
a waveguide film comprising a second optical material having a refractive index n2 which is greater than n1, said waveguide film disposed on said first planar surface of said substrate; and
capture molecules associated with said waveguide film and arranged in a plurality of discrete reaction sites on a surface thereof, each of said capture molecules capable of interacting selectively with at least one type of selected analyte molecule. - View Dependent Claims (53, 54, 55, 57, 58, 59, 60, 61, 62, 63)
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56. A method for performing a specific binding assay, the method comprising:
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providing a composite waveguide comprising a substrate, a waveguide film secured to said substrate, and a plurality of capture molecules on a surface of said waveguide film, opposite said substrate;
exposing said capture molecules to a solution including a sample that may comprise molecules of at least one selected analyte;
adding tracer molecules to said solution, each tracer molecule including a site capable of binding with at least a portion of a complementary capture molecule or at least a portion of said analyte, each tracer molecule including a component that emits fluorescent radiation of an emission wavelength when exposed to radiation of an excitation wavelength;
introducing light of said excitation wavelength into said waveguide film; and
detecting light of said emission wavelength passing through said substrate.
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Specification