Scattered total internal reflectance immunoassay system
First Claim
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1. An apparatus for detecting the presence of an analyte of interest inn a sample which comprises:
- a light source;
housing means for receiving an optically transparent member having a sample contacting surface, said member in said housing means being disposed such that the sample contacting surface is illuminated with light emitted from said light source; and
photodetection means which excludes the detection of light which propagates in a geometric optical path from the light source, said photodetection means being capable of detecting elastically-scattered light which propagates through the optically transparent member from the illuminated sample contacting surface between the plane of the sample contacting surface and the total internal reflectance critical angle of the sample contacting surface;
said photodetection means comprising a rotatable optical means comprising a first reflecting means and a second reflecting means whose optical axis is nearly coincident with the optical axis of said first reflecting means, whereby said elastically-scattered light is concentrated to facilitate detection thereof, said first reflecting means located to receive said elastically-scattered light and said second reflecting means located to receive light reflected from said first reflecting means.
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Abstract
A new immunoassay system is provided for the detection of ligands or ligand binding partners in solution in a heterogeneous format. The invention relies upon the detection of back scattered light from an evanescent wave disturbed by the presence of a colloidal gold label brought to the interface by an immunological reaction. The evanescent wave existing at the interface in turn is the result of a totally internally reflected incident light wave. Placement of the detector at a back angle above the critical angle insures a superior signal-to-noise ratio.
130 Citations
19 Claims
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1. An apparatus for detecting the presence of an analyte of interest inn a sample which comprises:
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a light source; housing means for receiving an optically transparent member having a sample contacting surface, said member in said housing means being disposed such that the sample contacting surface is illuminated with light emitted from said light source; and photodetection means which excludes the detection of light which propagates in a geometric optical path from the light source, said photodetection means being capable of detecting elastically-scattered light which propagates through the optically transparent member from the illuminated sample contacting surface between the plane of the sample contacting surface and the total internal reflectance critical angle of the sample contacting surface;
said photodetection means comprising a rotatable optical means comprising a first reflecting means and a second reflecting means whose optical axis is nearly coincident with the optical axis of said first reflecting means, whereby said elastically-scattered light is concentrated to facilitate detection thereof, said first reflecting means located to receive said elastically-scattered light and said second reflecting means located to receive light reflected from said first reflecting means.
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2. An apparatus for detecting the presence of an analyte of interest in a sample which comprises:
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a light source; housing means for receiving an optically transparent member having a sample contacting surface, said member in said housing means being disposed such that the sample contacting surface is illuminated with light emitted from said light source which propagates through the optically transparent member at angles between the plane of the sample contacting surface and the critical angle for total internal reflectance; and photodetection means which excludes the detection of light which propagates in a geometric optical path from the light source, said photodetection means being capable of detecting elastically--scattered light which propagates through the optically transparent member from the illuminates sample contacting surface between the plane of the sample contacting surface and the total internal reflectance critical angle;
said photodetection means comprising a rotatable optical means comprising a first reflecting means and a second reflecting means whose optical axis is nearly coincident with the optical axis of said first reflecting means, whereby said elastically-scattered light is concentrated to facilitate detection thereof, said first reflecting means located to receive said elastically-scattered light and said second reflecting means located to receive light reflected from said first reflecting means. - View Dependent Claims (3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15)
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16. A method for detecting the presence of a light scattering molecule on the surface of an optically transparent material which comprises illuminating said light scattering molecule from light through rotating optical reflectors detecting light scattered elastically by said light scattering molecule which propagates through said optically transparent material between the plane of the surface of the optically transparent material on which the light scattering molecule is located and the total internal reflectance critical angle of the surface on which the light scattering molecule is located, and correlating detected, elastically-scattered light to the presence of the light scattering molecule on the surface of the optically transparent material.
- 17. A method for detecting the presence of a light scattering molecule on the surface of an optically transparent material which comprises illuminating said light scattering molecule with an evanescent wave resulting from a light wave which propagates through said optically transparent material from light through rotating optical reflectors, detecting light scattered elastically by said light scattering molecule which propagates through said optically transparent material between the plane of the surface of the optically transparent material on which the light scattering molecule is located and the total internal reflectance critical angle of the surface on which the light scattering molecule is located, and correlating detected, elastically-scattered light to the presence of the light scattering molecule on the surface of the optically transparent material.
Specification