Refractomer and method for qualitative and quantitative measurements
First Claim
1. A method of using critical angle refractometry for sensing presence or absence of an analyte at a binding layer, the method comprising:
- providing a first optically transparent element and a second optically transparent element, the first optically transparent element having a higher refractive index than that of the second optically transparent element, the second element having the binding layer;
providing a contacting phase;
allowing the contacting phase to contact the binding layer of the second optically transparent element;
passing light through the first and the second optically transparent elements to cause the light to impinge upon an interface between the second optically transparent element and the binding layer; and
detecting a location of a boundary between a light area and a dark area on a sensing element, the location of the boundary being indicative of the presence or absence of the ligands at the binding layer.
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Accused Products
Abstract
A sensor apparatus and associated method for sensing and monitoring specific binding of analyte to an immobilized binding layer are disclosed. The apparatus preferably comprises an automatic critical angle refractometer having a linear scanned array and an optical system for illuminating a portion of the array, which illumination depends upon the refractive index of the binding layer deposited on an optically transparent element. The apparatus further includes a flow cell for bringing the analyte in contact with the binding layer. The apparatus also includes a computer for receiving and processing refractive index data from the critical angle refractometer during the reaction between the analyte and the layer, which computer may be peripherally connected to the refractometer or enclosed within the refractometer housing. A preferred sensing method of the present invention generally comprises providing a critical angle refractometer generating light impinging upon the immobilized binding layer, contacting the binding layer with a contacting phase, measuring the critical angle of total reflection, which measurements are indicative of the presence or absence of interactions between the analyte and the binding layer.
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Citations
30 Claims
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1. A method of using critical angle refractometry for sensing presence or absence of an analyte at a binding layer, the method comprising:
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providing a first optically transparent element and a second optically transparent element, the first optically transparent element having a higher refractive index than that of the second optically transparent element, the second element having the binding layer;
providing a contacting phase;
allowing the contacting phase to contact the binding layer of the second optically transparent element;
passing light through the first and the second optically transparent elements to cause the light to impinge upon an interface between the second optically transparent element and the binding layer; and
detecting a location of a boundary between a light area and a dark area on a sensing element, the location of the boundary being indicative of the presence or absence of the ligands at the binding layer. - View Dependent Claims (2)
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3. A method of using critical angle refractometry for sensing presence or absence of an analyte at a binding layer of a first optically transparent material, the method comprising:
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providing the first optically transparent material of a higher optical density than that of the binding layer;
contacting the binding layer with a contacting phase;
passing light along an optical path through the first optically transparent material to cause the light to impinge upon an interface between the binding layer and the first optically transparent material;
sensing a boundary between a light area and a dark area on a sensing element disposed along the optical path; and
utilizing the location of the boundary to determine the presence or absence of the analyte at the binding layer. - View Dependent Claims (4, 5)
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6. A method of sensing presence or absence of an analyte at a binding layer, the method comprising:
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providing an interface between the binding layer and an optically transparent element, the interface being located along an optical path, the binding layer and the optically transparent element having different optical densities sufficient to totally internally reflect light impinging on the interface;
contacting the binding layer with a contacting phase;
illuminating the interface with the light propagating along the optical path, so that a portion of the light totally internally reflected from the interface propagates between the interface and a sensing element disposed along the optical path and illuminates the sensing element to form a light area thereon; and
detecting a location of a boundary between the light area and a dark area on the sensing element, the location of the boundary being indicative of the presence or absence of the analyte at the binding layer. - View Dependent Claims (7, 8, 9)
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10. A method of sensing presence or absence of an analyte at a binding layer, the method comprising:
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providing a first interface between a first medium and a second medium, a second interface between the second medium and a third medium, and a third interface between the third medium and the binding layer, the first, the second, the third media and the binding layer having optical densities sufficient to cause light propagating through the first, the second and the third interfaces to be totally internally reflected at the third interface and to propagate toward a sensor;
contacting the binding layer with a contacting phase;
passing the light through the first interface and the second interfaces to cause the light to impinge upon the third interface; and
detecting a location of a boundary between a dark area and a light area on the sensor, the location of the boundary being indicative of the presence or absence of the ligands at the binding layer. - View Dependent Claims (11)
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12. A method of sensing presence or absence of an analyte at a binding layer, the method comprising:
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providing a light beam generated by a light source;
providing an interface between the binding layer and an optically transparent element, the binding layer and the optically transparent element having optical densities sufficient to cause the light beam impinging upon the interface to be totally internally reflected;
contacting the binding layer with a contacting phase;
illuminating the interface by the light beam impinging upon the interface at a predetermined angle of incidence;
providing a sensor located at a position in which the sensor can sense the light totally internally reflected at the interface; and
sensing the presence or absence of a boundary between a dark area and a light area on the sensor, the presence or absence of the boundary being indicative of the presence or absence of the analyte at the binding layer. - View Dependent Claims (13, 14)
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15. A method of sensing an analyte at a binding layer, the method comprising:
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providing an optically transparent element having an optical density different from that of the binding layer, the optically transparent element comprising the interface between the binding layer and the optically, transparent element;
providing a light source generating a collimated light beam propagating along an optical path, the optical path comprising the light source, the interface and a sensing element;
contacting the binding layer with a contacting phase;
directing the light beam to the interface to cause the light beam to impinge upon the interface at a predetermined angle of incidence; and
sensing presence or absence of a boundary between a dark area and a light area on the sensing element to determine the absence of the analyte in the contacting phase.
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16. A method of using critical angle refractometry for determining presence or absence of an analyte at a binding layer deposited on an optically transparent element, the method comprising:
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providing a critical angle refractometer defining an optical path;
the critical angle refractometer comprising a sensing element disposed along the optical path;
directing light along the optical path at an interface between the binding layer and the optically transparent element;
bringing the analyte in contact with the binding layer; and
refractometrically detecting changes in an optical density of the binding layer by sensing the light on the sensing element; and
and relating the changes to the presence or absence of the analyte at the binding layer.- View Dependent Claims (17, 18, 19)
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20. An apparatus for using critical angle refractometry to sense presence or absence of an analyte at a binding layer, the analyte having affinity to the binding layer, the apparatus comprising:
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a first optically transparent element and a second optically transparent element, the first optically transparent element having a higher refractive index than that of the second optically transparent element;
the binding layer deposited on the second optically transparent element, the binding layer having a refractive index lower than that of the second optically transparent element;
a contacting phase contacting the binding layer;
a light beam passing through the first and the second optically transparent elements, causing the light to impinge upon an interface between the second optically transparent element and the binding layer; and
a sensor detecting a location of a boundary between a light area and a dark area on the sensor. - View Dependent Claims (21, 22, 23)
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24. A system for determining presence or absence of an analyte in a contacting phase, the system comprising:
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a critical angle refractometer defining an optical path of a light beam impinging upon an interface between an optically transparent element and a binding layer, the binding layer being deposited on the optically transparent element and having affinity to the analyte;
the contacting phase contacting the binding layer; and
a sensor sensing a boundary between light and dark areas on the sensor. - View Dependent Claims (25, 26, 27)
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28. A system for detecting presence or absence of an analyte in a contacting phase, the system comprising:
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an optically transparent element having a binding layer deposited thereon, the binding layer having affinity to the analyte;
a critical angle refractometer defining an optical path of a collimated light beam impinging upon an interface between the binding layer and the optically transparent element;
the contacting phase contacting the binding layer; and
a sensor disposed along the optical path to detect changes in an optical density of the binding layer by sensing the collimated light beam travelling along the optical path. - View Dependent Claims (29, 30)
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Specification