Immunoassay apparatus
First Claim
1. In apparatus for assaying a fluid sample and including a totally internally reflecting, elongated, substrate transmissive to excitation radiation directed at a proximal end of said substrate by an optical system;
- which radiation, when propagated through said substrate, will provide an evanescent wave for exciting fluorescence in fluorescent material disposed at least on a portion of the surface of said substrate, said substrate also being transmissive to said fluorescence; and
a hollow elongated enclosure spaced from and surrounding the surface of said substrate, the improvement comprising;
mounting means couplable to said optical system and to said substrate and to said enclosure for mounting said substrate within said enclosure, said mounting means comprising;
seat means spaced a fixed predetermined distance from said optical system when said mounting means is coupled to said optical system, for so releasably seating said proximal end in a preselected position fixed at least axially with respect to the long axis of said substrate and with respect to said optical system, that when said proximal end is seated in said seat means, substantially all of said excitation radiation entering said proximal end from said optical system at a solid angle less than or equal to a maximum acceptance angle determined in part by the refractive index of said sample, undergoes total internal reflection within said substrate.
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Accused Products
Abstract
An assay apparatus employing total internal reflection of excitation radiation at the interface between a replaceable optically conductive rod or fiber and a surrounding liquid phase of lower index of refraction. Immobilized on the surface of the fiber is a component of a complex formed in an immunological-type specific reaction. A fluorophore that can be excited into fluorescence by the excitation radiation is attached to another component of the complex. The rod is coaxially mounted within a tube that is sized with respect to said rod so that a fluid sample may be introduced into said tube.
The rod and tubing are supported in a mounting assembly that is attachable to an optical assembly for transmitting excitation radiation into the proximal end of the rod and receiving fluorescent radiation emitted from the proximal end of the rod. Included in the apparatus is a mounting assembly for centering the rod within tube and for biasing the rod in a first direction against an annular seat. The latter is designed to support one end of the rod so that none of the radiation introduced into the rod is intercepted by the seat. The apparatus also includes a seal arrangement for preventing a fluid sample from escaping from the tube. Additionally, a tapered fiber may be included in the optical system of the assay apparatus for increasing the power and numerical aperture of radiation input into the optical rod.
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Citations
32 Claims
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1. In apparatus for assaying a fluid sample and including a totally internally reflecting, elongated, substrate transmissive to excitation radiation directed at a proximal end of said substrate by an optical system;
- which radiation, when propagated through said substrate, will provide an evanescent wave for exciting fluorescence in fluorescent material disposed at least on a portion of the surface of said substrate, said substrate also being transmissive to said fluorescence; and
a hollow elongated enclosure spaced from and surrounding the surface of said substrate, the improvement comprising;mounting means couplable to said optical system and to said substrate and to said enclosure for mounting said substrate within said enclosure, said mounting means comprising; seat means spaced a fixed predetermined distance from said optical system when said mounting means is coupled to said optical system, for so releasably seating said proximal end in a preselected position fixed at least axially with respect to the long axis of said substrate and with respect to said optical system, that when said proximal end is seated in said seat means, substantially all of said excitation radiation entering said proximal end from said optical system at a solid angle less than or equal to a maximum acceptance angle determined in part by the refractive index of said sample, undergoes total internal reflection within said substrate. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 28, 29, 32)
- which radiation, when propagated through said substrate, will provide an evanescent wave for exciting fluorescence in fluorescent material disposed at least on a portion of the surface of said substrate, said substrate also being transmissive to said fluorescence; and
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13. In apparatus for assaying a fluid sample and including a totally internally reflecting elongated substrate transmissive to radiation capable of providing an evanescent wave for exciting fluorescence in fluorescent material disposed at leas on a portion of the surface of said substrate, said substrate also being transmissive to said fluorescence, and elongated means spaced from said surface of said substrate so as to define a hollow elongated enclosure surrounding said surface, and optical means couplable to said mounting means for transmitting a beam of optical radiation to an end face of said substrate, the improvement comprising:
means coupled to said substrate and said elongated means for mounting said substrate within said enclosure and for releasably supporting both ends of said elongated substrate so that said end face is axially positionable at a fixed location with respect to said optical means and so that said elongated substrate is (a) substantially prevented from moving transversely relative to its longitudinal axis and (b) is prevented from moving in a first axial direction relative to said optical means. - View Dependent Claims (14, 15, 16, 17, 18, 19, 20, 21, 22)
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23. In apparatus for assaying a fluid sample and including a totally internally reflecting, elongated, substrate transmissive to excitation radiation from a radiation source, which radiation is capable of providing an evanescent wave for exciting fluorescence in fluorescent material disposed at least on a portion of the surface of said substrate, said substrate also being transmissive to said fluorescence, said substrate comprising a proximal end through which said excitation radiation and excited fluorescence may be transmitted, a hollow elongated enclosure spaced from and surrounding the surface of said substrate, the improvement comprising:
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mounting means couplable to said substrate and to said enclosure for mounting said substrate within said enclosure, said mounting means comprising; means defining a fixed interface between said mounting means and said radiation source; seat means for supporting said proximal end in a preselected position with respect to said interface, so that substantially all of said excitation radiation entering said proximal end at a solid angle less than or equal to a maximum acceptance angle determined in part by the refractive index of said sample undergoes total internal reflection within said substrate; and seal means for creating a barrier past which a fluid sample disposed within said enclosure cannot escape. - View Dependent Claims (24, 25, 26, 27)
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30. In apparatus for assaying a fluid sample and including a totally internally reflecting elongated substrate transmissive to radiation capable of providing an evanescent wave for exciting fluorescence in fluorescent material disposed at least on a portion of the surface of said substrate, said substrate also being transmissive to said fluorescence, and elongated means spaced from said surface of said substrate so as to define a hollow elongated enclosure surrounding said surface, and optical means couplable to said mounting means for transmitting a beam of optical radiation to an end face of said substrate, the improvement comprising:
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means coupled to said substrate and said elongated means for mounting said substrate within said enclosure and for releasably supporting both ends of said elongated substrate so that said end face is axially positionable at a fixed location with respect to said optical means and so that said elongated substrate is (a) substantially prevented from moving transversely relative to its longitudinal axis and (b) is prevented from moving in a first axial direction relative to said optical means; and wherein said optical means comprises a tapered fiber disposed in fixed spatial relationship relative to said fiber end face, said tapered fiber having an input face and an output face, said tapered fiber being transmissive to said optical radiation and being tapered smoothly so as to reduce the diameter of said tapered fiber from said input face to said output face. - View Dependent Claims (31)
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Specification