CA2427689A1 - A label-free high-throughput optical technique for detecting biomolecular interactions - Google Patents
A label-free high-throughput optical technique for detecting biomolecular interactions Download PDFInfo
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- CA2427689A1 CA2427689A1 CA002427689A CA2427689A CA2427689A1 CA 2427689 A1 CA2427689 A1 CA 2427689A1 CA 002427689 A CA002427689 A CA 002427689A CA 2427689 A CA2427689 A CA 2427689A CA 2427689 A1 CA2427689 A1 CA 2427689A1
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54366—Apparatus specially adapted for solid-phase testing
- G01N33/54373—Apparatus specially adapted for solid-phase testing involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/508—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above
- B01L3/5085—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above for multiple samples, e.g. microtitration plates
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/251—Colorimeters; Construction thereof
- G01N21/253—Colorimeters; Construction thereof for batch operation, i.e. multisample apparatus
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/47—Scattering, i.e. diffuse reflection
- G01N21/4788—Diffraction
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N21/7703—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator using reagent-clad optical fibres or optical waveguides
- G01N21/774—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator using reagent-clad optical fibres or optical waveguides the reagent being on a grating or periodic structure
- G01N21/7743—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator using reagent-clad optical fibres or optical waveguides the reagent being on a grating or periodic structure the reagent-coated grating coupling light in or out of the waveguide
-
- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B5/00—Optical elements other than lenses
- G02B5/18—Diffraction gratings
- G02B5/1809—Diffraction gratings with pitch less than or comparable to the wavelength
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
- Y10T436/00—Chemistry: analytical and immunological testing
- Y10T436/24—Nuclear magnetic resonance, electron spin resonance or other spin effects or mass spectrometry
Abstract
Published without an
Claims (104)
1. A biosensor comprising:
(a) a two-dimensional grating comprised of a material having a high refractive index;
(b) a substrate layer that supports the two-dimensional grating; and (c) one or more specific binding substances immobilized on the surface of the two-dimensional grating opposite of the substrate layer;
wherein, when the biosensor is illuminated a resonant grating effect is produced on the reflected radiation spectrum, and wherein the depth and period of the two-dimensional grating are less than the wavelength of the resonant grating effect.
(a) a two-dimensional grating comprised of a material having a high refractive index;
(b) a substrate layer that supports the two-dimensional grating; and (c) one or more specific binding substances immobilized on the surface of the two-dimensional grating opposite of the substrate layer;
wherein, when the biosensor is illuminated a resonant grating effect is produced on the reflected radiation spectrum, and wherein the depth and period of the two-dimensional grating are less than the wavelength of the resonant grating effect.
2. The biosensor of claim 1, wherein a narrow band of optical wavelengths is reflected from the biosensor when the biosensor is illuminated with a broad band of optical wavelengths.
3. The biosensor of claim 1, wherein the substrate comprises glass, plastic or epoxy.
4. The biosensor of claim 1, wherein the two-dimensional grating is comprised of a material selected from the group consisting of zinc sulfide, titanium dioxide, tantalum oxide, and silicon nitride.
5. The biosensor of claim 1, further comprising a cover layer on the surface of the two-dimensional grating opposite of the substrate layer, wherein the one or more specific binding substances are immobilized on the surface of the cover layer opposite of the two-dimensional grating.
6. The biosensor of claim 5, wherein the cover layer comprises a material that has a lower refractive index than zinc sulfide, titanium dioxide, tantalum oxide, and silicon nitride.
7. The biosensor of claim 6, wherein the cover layer comprises a material selected from the group consisting of glass, epoxy, and plastic.
8. The biosensor of claim 1, wherein the two-dimensional grating has a period of about 0.01 microns to about 1 micron and a depth of about 0.01 microns to about 1 micron.
9. The biosensor of claim 1, wherein the one or more specific binding substances are arranged in an array of distinct locations.
10. The biosensor of claim 1, wherein the one or more specific binding substances are immobilized on the two-dimensional grating by physical adsorption or by chemical binding.
11. The biosensor of claim 9, wherein the distinct locations define a microarray spot of about 50-500 microns in diameter.
12. The biosensor of claim 1, wherein the one or more specific binding substances are bound to their binding partners.
13. The biosensor of claim 1, wherein the one or more specific binding substances are selected from the group consisting of nucleic acids, polypeptides, antigens, polyclonal antibodies, monoclonal antibodies, single chain antibodies (scFv), F(ab) fragments, F(ab')2 fragments, Fv fragments, small organic molecules, cells, viruses, bacteria, and biological samples.
14. The biosensor of claim 13, wherein the biological sample is selected from the group consisting of blood, plasma, serum, gastrointestinal secretions, homogenates of tissues or tumors, synovial fluid, feces, saliva, sputum, cyst fluid, amniotic fluid, cerebrospinal fluid, peritoneal fluid, lung lavage fluid, semen, lymphatic fluid, tears, and prostatitc fluid.
15. The biosensor of claim 12, wherein the binding partners are selected from the group consisting of nucleic acids, polypeptides, antigens, polyclonal antibodies, monoclonal antibodies, single chain antibodies (scFv), F(ab) fragments, F(ab')2 fragments, Fv fragments, small organic molecules, cells, viruses, bacteria, and biological samples.
16. The biosensor of claim 15, wherein the biological sample is selected from the group consisting of blood, plasma, serum, gastrointestinal secretions, homogenates of tissues or tumors, synovial fluid, feces, saliva, sputum, cyst fluid, amniotic fluid, cerebrospinal fluid, peritoneal fluid, lung lavage fluid, semen, lymphatic fluid, tears, and prostatitc fluid.
17. A liquid-containing vessel comprising the biosensor of claim 1 as an internal surface.
18. The liquid-containing vessel of claim 17, wherein the vessel is selected from the group consisting of a microtiter plate, a test tube, a petri dish and a microfluidic channel.
19. A detection system comprising the biosensor of claim 1, a light source that directs light to the biosensor, and a detector that detects light reflected from the biosensor, wherein a polarizing filter occurs between the light source and the biosensor.
20. A method of detecting the binding of one or more specific binding substances to their respective binding partners comprising:
(a) applying one or more binding partners to the biosensor of claim 1;
(b) illuminating the biosensor with light; and (c) detecting a maxima in reflected wavelength, or a minima in transmitted wavelength of light from the biosensor;
wherein, if the one or more specific binding substances have bound to their respective binding partners, then the reflected wavelength of light is shifted.
(a) applying one or more binding partners to the biosensor of claim 1;
(b) illuminating the biosensor with light; and (c) detecting a maxima in reflected wavelength, or a minima in transmitted wavelength of light from the biosensor;
wherein, if the one or more specific binding substances have bound to their respective binding partners, then the reflected wavelength of light is shifted.
21. A method of detecting the binding of one or more specific binding substances to their respective binding partners comprising:
(a) applying one or more binding partners to the biosensor of claim 1, wherein the two-dimensional grating is coated with an array of distinct locations containing the one or more specific binding substances;
(b) illuminating each distinct location of the biosensor with light; and (c) detecting maximum reflected wavelength or minimum transmitted wavelength of light from each distinct location of the biosensor;
wherein, if the one or more specific binding substances have bound to their respective binding partners at a distinct location, then the reflected wavelength of light is shifted.
(a) applying one or more binding partners to the biosensor of claim 1, wherein the two-dimensional grating is coated with an array of distinct locations containing the one or more specific binding substances;
(b) illuminating each distinct location of the biosensor with light; and (c) detecting maximum reflected wavelength or minimum transmitted wavelength of light from each distinct location of the biosensor;
wherein, if the one or more specific binding substances have bound to their respective binding partners at a distinct location, then the reflected wavelength of light is shifted.
22. A method of detecting activity of an enzyme comprising:
(a) applying one or more enzymes to the biosensor of claim 1;
(b) washing the biosensor;
(c) illuminating the biosensor with light; and (d) detecting reflected wavelength of light from the biosensor;
wherein, if the one or more enzymes have altered the one or more specific binding substances of the biosensor by enzymatic activity, then the reflected wavelength of light is shifted.
(a) applying one or more enzymes to the biosensor of claim 1;
(b) washing the biosensor;
(c) illuminating the biosensor with light; and (d) detecting reflected wavelength of light from the biosensor;
wherein, if the one or more enzymes have altered the one or more specific binding substances of the biosensor by enzymatic activity, then the reflected wavelength of light is shifted.
23. A biosensor comprising:
(a) a sheet material having a first and second surface, wherein the first surface defines relief volume diffraction structures;
(b) a reflective material coated onto the first surface of the sheet material;
and (c) one or more specific binding substances immobilized on the reflective material;
wherein the biosensor reflects light predominantly at a first single optical wavelength when illuminated with a broad band of optical wavelengths, and wherein the biosensor reflects light at a second single optical wavelength when the one or more specific binding substances are immobilized on the reflective surface, wherein the reflection at the second optical wavelength of light results from optical interference.
(a) a sheet material having a first and second surface, wherein the first surface defines relief volume diffraction structures;
(b) a reflective material coated onto the first surface of the sheet material;
and (c) one or more specific binding substances immobilized on the reflective material;
wherein the biosensor reflects light predominantly at a first single optical wavelength when illuminated with a broad band of optical wavelengths, and wherein the biosensor reflects light at a second single optical wavelength when the one or more specific binding substances are immobilized on the reflective surface, wherein the reflection at the second optical wavelength of light results from optical interference.
24. The biosensor of claim 23, wherein the biosensor reflects light at a third single optical wavelength when the one or more specific binding substances are bound to their respective binding partners, wherein the reflection at the third optical wavelength results from optical interference.
25. The biosensor of claim 23, wherein the depth and period of the relief volume diffraction structures are less than the resonance wavelength of the light reflected from the biosensor.
26. The biosensor of claim 23, wherein the relief volume diffraction structures have a period of about 0.01 microns to about 1 micron and a depth of about 0.01 micron to about 1 micron.
27. The biosensor of claim 23, wherein the one or more specific binding substances are bound to their respective binding partners.
28. A liquid-containing vessel comprising the biosensor of claim 23 as an internal surface.
29. The liquid-containing vessel of claim 28, wherein the vessel is selected from the group consisting of a microtiter plate, a test tube, a petri dish and a microfluidic channel.
30. The biosensor of claim 23, wherein the one or more specific binding substances are arranged in an array of distinct locations on the reflective material.
31. The biosensor of claim 30, wherein the distinct locations define a microarray spot of about 50-500 microns in diameter.
32. The biosensor of claim 23, wherein the one or more specific binding substances are immobilized to the reflective material by physical adsorption or by chemical binding.
33. The biosensor of claim 32, wherein the relief volume diffraction structures are about 0.5 microns to about 5 microns in diameter.
34. A method of detecting the binding of one or more specific binding substances to their respective binding partners comprising:
(a) applying one or more binding partners to the biosensor of claim 23;
(b) illuminating the biosensor with light; and (c) detecting reflected wavelength of light from the biosensor;
wherein, if the one or more specific binding substances have bound to their respective binding partners, then the reflected wavelength of light is shifted.
(a) applying one or more binding partners to the biosensor of claim 23;
(b) illuminating the biosensor with light; and (c) detecting reflected wavelength of light from the biosensor;
wherein, if the one or more specific binding substances have bound to their respective binding partners, then the reflected wavelength of light is shifted.
35. A method of detecting the binding of one or more specific binding substances to their respective binding partners comprising:
(a) applying one or more binding partners to the biosensor of claim 23, wherein the one or more specific binding substances are arranged in an array of distinct locations on the reflective material;
(b) illuminating each distinct location of the biosensor with light; and (c) detecting reflected wavelength of light from each distinct location of the biosensor;
wherein, if the one or more specific binding substances have bound to their respective binding partners at a distinct location, then the reflected wavelength of light is shifted.
(a) applying one or more binding partners to the biosensor of claim 23, wherein the one or more specific binding substances are arranged in an array of distinct locations on the reflective material;
(b) illuminating each distinct location of the biosensor with light; and (c) detecting reflected wavelength of light from each distinct location of the biosensor;
wherein, if the one or more specific binding substances have bound to their respective binding partners at a distinct location, then the reflected wavelength of light is shifted.
36. A method of detecting activity of an enzyme comprising:
(a) applying one or more enzymes to the biosensor of claim 23;
(b) washing the biosensor;
(c) illuminating the biosensor with light; and (d) detecting reflected wavelength of light from the biosensor;
wherein, if the one or more enzymes have altered the one or more specific binding substances of the biosensor by enzymatic activity, then the reflected wavelength of light is shifted.
(a) applying one or more enzymes to the biosensor of claim 23;
(b) washing the biosensor;
(c) illuminating the biosensor with light; and (d) detecting reflected wavelength of light from the biosensor;
wherein, if the one or more enzymes have altered the one or more specific binding substances of the biosensor by enzymatic activity, then the reflected wavelength of light is shifted.
37. A biosensor comprising a two-dimensional grating having a first and a second surface comprised of an optically transparent material that conducts electricity, wherein the first surface of the two-dimensional grating is coated with an electrical insulator, and wherein the second surface of the two-dimensional grating is deposited on a substrate, wherein when the biosensor is illuminated a resonant grating effect is produced on the reflected radiation spectrum, wherein the depth and the period of the two-dimensional grating are less than the wavelength of the resonant grating effect.
38. The biosensor of claim 37, wherein the two-dimensional grating is comprised of a repeating pattern of shapes selected from the group consisting of squares, circles, ellipses, triangles, ovals, trapezoids, sinusoidal waves, rectangles, and hexagons.
39. The biosensor of claim 37, wherein the repeating pattern of shapes are arranged in a rectangular grid or hexagonal grid.
40. The biosensor of claim 37, wherein the two-dimensional grating has a period of about 0.01 microns to about 1 micron and a depth of about 0.01 microns to about 1 micron.
41. The biosensor of claim 37, wherein two or more separate grating regions are present on the same substrate.
42. The biosensor of claim 41, further comprising an electrically conducting trace to each separate grating region of the substrate.
43. The biosensor of claim 42, wherein the conducting trace is connected to a voltage source.
44. The biosensor of claim 41, wherein one or more specific binding substances are bound to each separate grating region of the substrate.
45. The biosensor of claim 44, wherein the one or more specific binding substances are bound to their respective binding partners.
46. A liquid-containing vessel comprising the biosensor of claim 37 as an internal surface.
47. The liquid-containing vessel of claim 46, wherein the vessel is selected from the group consisting of a microtiter plate, a test tube, a petri dish and a microfluidic channel.
48. A method of detecting the binding of one or more specific binding substances to their respective binding partners comprising:
(a) applying one or more binding partners to the biosensor of claim 37;
(b) applying an electrical charge to the electrically conducting traces;
(c) illuminating the biosensor with light; and (d) detecting reflected wavelength of light from the biosensor;
wherein, if the one or more specific binding substances have bound to their respective binding partners, then the reflected wavelength of light is shifted.
(a) applying one or more binding partners to the biosensor of claim 37;
(b) applying an electrical charge to the electrically conducting traces;
(c) illuminating the biosensor with light; and (d) detecting reflected wavelength of light from the biosensor;
wherein, if the one or more specific binding substances have bound to their respective binding partners, then the reflected wavelength of light is shifted.
49. The method of claim 48, further comprising the step of applying a reversed electrical charge to the electrically conducting traces before illuminating the biosensor with light.
50. A method of measuring the amount of one or more binding partners in a test sample comprising:
(a) illuminating the biosensor of claims 1, 23, or 37 with light;
(b) detecting reflected wavelength of light from the biosensor;
(d) applying a test sample comprising one or more binding partners to the biosensor;
(e) illuminating the biosensor with light; and (f) detecting reflected wavelength of light from the biosensor;
wherein, the difference in wavelength of light in step (b) and step (f) is a measurement of the amount of one or more binding partners in the test sample.
(a) illuminating the biosensor of claims 1, 23, or 37 with light;
(b) detecting reflected wavelength of light from the biosensor;
(d) applying a test sample comprising one or more binding partners to the biosensor;
(e) illuminating the biosensor with light; and (f) detecting reflected wavelength of light from the biosensor;
wherein, the difference in wavelength of light in step (b) and step (f) is a measurement of the amount of one or more binding partners in the test sample.
51. A detection system comprising the biosensor of claims 1, 23, or 37, a light source that directs light at the biosensor, and a detector that detects light reflected from the biosensor, wherein a first illuminating fiber probe having two ends is connected at its first end to the detector, wherein a second collection fiber probe having two ends is connected at its first end to the light source, wherein the first and second fiber probes are connected at their second ends to a third fiber probe, wherein the third fiber probe acts as an illumination and collection fiber probe, and wherein the third fiber probe is oriented at a normal angle of incidence to the biosensor and supports counter-propagating illuminating and reflecting optical signals.
52. A detection system comprising the biosensor of claims 1, 23, or 37, a light source that directs light at the biosensor, and a detector that detects light reflected from the biosensor, wherein an illuminating fiber probe is connected to the light source and is oriented at a 90 degree angle to a collecting fiber probe, wherein the collecting fiber probe is connected to the detector, wherein light is directed through the illuminating fiber probe into a beam splitter that directs the light to the biosensor, wherein reflected light is directed into the beam splitter that directs the light into the collecting fiber.
53. A method of immobilizing one or more specific binding substances onto the biosensor of claim 1, 23, or 37 comprising activating the biosensor with amine, attaching linker groups to the amine-activated biosensor, and attaching one or more specific binding substances to the linker groups.
54. The method of claim 53, wherein the biosensor is activated with amine by a method comprising:
(a) immersing the biosensor into a piranha solution;
(b) washing the biosensor;
(c) immersing the biosensor in 3% 3-aminopropyltriethoxysilane solution in dry acetone;
(d) washing the biosensor in dry acetone; and (e) washing the biosensor with water.
(a) immersing the biosensor into a piranha solution;
(b) washing the biosensor;
(c) immersing the biosensor in 3% 3-aminopropyltriethoxysilane solution in dry acetone;
(d) washing the biosensor in dry acetone; and (e) washing the biosensor with water.
55. The method of claim 53, wherein the linker is selected from the group consisting of amine;
aldehyde; N, N'-disuccinimidyl carbonate; and nickel.
aldehyde; N, N'-disuccinimidyl carbonate; and nickel.
56. A method of detecting the binding of one or more specific binding substances to their respective binding partners comprising:
(a) applying one or more binding partners comprising one or more tags to the biosensor of claims 1, 23, or 37;
(b) illuminating the biosensor with light; and (c) detecting reflected wavelength of light from the biosensor;
wherein, if the one or more specific binding substances have bound to their respective binding partners, then the reflected wavelength of light is shifted.
(a) applying one or more binding partners comprising one or more tags to the biosensor of claims 1, 23, or 37;
(b) illuminating the biosensor with light; and (c) detecting reflected wavelength of light from the biosensor;
wherein, if the one or more specific binding substances have bound to their respective binding partners, then the reflected wavelength of light is shifted.
57. The method of claim 56, wherein the one or more tags are selected from the group consisting of biotin, succinimidyl-6-[a-methyl-a-(2-pyridyl-dithio) toluamido]
hexanoate (SMPT), dimethylpimelimidate (DMP), and histidine.
hexanoate (SMPT), dimethylpimelimidate (DMP), and histidine.
58. The method of claim 56, wherein the one or more tags are reacted with a composition selected from the group consisting of streptavidin, horseradish peroxidase, and streptavidin coated nanoparticles, before the step of illuminating the biosensor with light.
59. A biosensor composition comprising two or more biosensors of claims 9, 30, or 44 wherein the biosensors are associated with a holding fixture.
60. The biosensor composition of claim 59, wherein the composition comprises about 50 to about 1,000 individual biosensors.
61. The biosensor composition of claim 59, wherein the composition comprises about 96 biosensors.
62. The biosensor composition of claim 59, wherein the composition comprises about 384 biosensors.
63. The biosensor composition of claim 59, wherein the two or more biosensors each comprise about 25 to about 1,000 distinct locations.
64. The biosensor composition of claim 59, wherein each biosensor is about 1 mm2 to about 5 mm2.
65. The biosensor composition of claim 59, wherein each biosensor is about 3 mm2.
66. The biosensor composition of claim 59, wherein the holding fixture holds each biosensor such that each biosensor can be placed into a separate well of a microtiter plate.
67. A biosensor composition comprising one or more biosensors of claims 1, 23, or 37 on a tip of a multi-fiber optic probe.
68. The biosensor composition of claim 67, wherein the one or more biosensors are fabricated into the tip of the probe.
69. The biosensor composition of claim 67, wherein the one or more biosensors are attached onto the tip of the probe.
70. A method of detecting binding of one or more specific binding substances to their respective binding partners in vivo comprising:
(a) inserting the tip of the fiber optic probe of claim 67 into the body of a human or animal;
(b) illuminating the biosensor with light;
(c) detecting reflected wavelength of light from the biosensor;
wherein, if the one or more specific binding substances have bound to their respective binding partners, then the reflected wavelength of light is shifted.
(a) inserting the tip of the fiber optic probe of claim 67 into the body of a human or animal;
(b) illuminating the biosensor with light;
(c) detecting reflected wavelength of light from the biosensor;
wherein, if the one or more specific binding substances have bound to their respective binding partners, then the reflected wavelength of light is shifted.
71. A detection system comprising:
(a) the biosensor of claim 1;
(b) a laser source that directs a laser beam to a scanning mirror device, wherein the scanning mirror device is used to vary the laser beam's incident angle;
(c) an optical system for maintaining columination of the incident laser beam;
(d) and a light detector.
(a) the biosensor of claim 1;
(b) a laser source that directs a laser beam to a scanning mirror device, wherein the scanning mirror device is used to vary the laser beam's incident angle;
(c) an optical system for maintaining columination of the incident laser beam;
(d) and a light detector.
72. The detection system of claim 71, wherein the scanning mirror device is a linear galvanometer.
73. The detection system of claim 72, wherein the linear galvanometer operates at a frequency of about 2 Hz to about 120 Hz and a mechanical scan angle of about 10 degrees to about 20 degrees.
74. The detection system of claim 71, wherein the laser is a diode laser with a wavelength selected from the group consisting of 780 nm, 785 nm, 810 nm, and 830 nm.
75. A method for determining a location of a resonant peak for a binding partner in a resonant reflectance spectrum with a colormetric resonant biosensor, comprising:
selecting a set of resonant reflectance data for a plurality of colormetric resonant biosensor distinct locations, wherein the set of resonant reflectance data is collected by illuminating a colormetric resonant diffractive grating surface with a light source and measuring reflected light at a pre-determined incidence, wherein the colormetric resonant diffractive grating surface is used as a surface binding platform for one or more specific binding substances, wherein binding partners can be detected without use of a molecular label, wherein the set of resonant reflectance data includes a plurality of sets of two measurements, where a first measurement includes a first reflectance spectra of one or more specific binding substances that are attached to the colormetric resonant diffractive grating surface and a second measurement includes a second reflectance spectra of the one or more specific binding substance after one or more binding partners are applied to colormetric resonant diffractive grating surface including the one or more specific binding substances, and wherein a difference in a peak wavelength between the first and second measurement is a measurement of an amount of binding partners that bound to the one or more specific binding substances;
determining a maximum value for a second measurement from the plurality of sets of two measurements from the set of resonant reflectance data for the plurality of binding partners, wherein the maximum value includes inherent noise included in the resonant reflectance data;
determining whether the maximum value is greater than a pre-determined threshold, and if so, defining a curve-fit region around the determined maximum value, performing a curve-fitting procedure to fit a curve around the curve-fit region, wherein the curve-fitting procedure removes a pre-determined amount of inherent noise included in the resonant reflectance data;
determining a location of a maximum resonant peak on the fitted curve; and determining a value of the maximum resonant peak, wherein the value of the maximum resonant peak is used to identify an amount of biomolecular binding of the one or more specific binding substances to the one or more binding partners.
selecting a set of resonant reflectance data for a plurality of colormetric resonant biosensor distinct locations, wherein the set of resonant reflectance data is collected by illuminating a colormetric resonant diffractive grating surface with a light source and measuring reflected light at a pre-determined incidence, wherein the colormetric resonant diffractive grating surface is used as a surface binding platform for one or more specific binding substances, wherein binding partners can be detected without use of a molecular label, wherein the set of resonant reflectance data includes a plurality of sets of two measurements, where a first measurement includes a first reflectance spectra of one or more specific binding substances that are attached to the colormetric resonant diffractive grating surface and a second measurement includes a second reflectance spectra of the one or more specific binding substance after one or more binding partners are applied to colormetric resonant diffractive grating surface including the one or more specific binding substances, and wherein a difference in a peak wavelength between the first and second measurement is a measurement of an amount of binding partners that bound to the one or more specific binding substances;
determining a maximum value for a second measurement from the plurality of sets of two measurements from the set of resonant reflectance data for the plurality of binding partners, wherein the maximum value includes inherent noise included in the resonant reflectance data;
determining whether the maximum value is greater than a pre-determined threshold, and if so, defining a curve-fit region around the determined maximum value, performing a curve-fitting procedure to fit a curve around the curve-fit region, wherein the curve-fitting procedure removes a pre-determined amount of inherent noise included in the resonant reflectance data;
determining a location of a maximum resonant peak on the fitted curve; and determining a value of the maximum resonant peak, wherein the value of the maximum resonant peak is used to identify an amount of biomolecular binding of the one or more specific binding substances to the one or more binding partners.
76. A computer readable medium having stored therein instructions for causing a processor to execute the method of claim 75.
77. The method of claim 75 wherein a sensitivity of a colormetric resonant biosensor is determined by a shift in a location of a resonant peak in the plurality of sets of two measurements in the set of resonant reflectance data.
78. The method of claim 75 wherein the step of selecting a set of resonant reflectance data includes selecting a set of resonant reflectance data:
x i and y i for i = 1, 2, 3, ...n, wherein x i is where a first measurement includes a first reflectance spectra of one or more specific binding substance attached to the colormetric resonant diffractive grating surface, y i a second measurement includes a second reflectance spectra of the one or more specific binding substances after a plurality of binding partners are applied to colormetric resonant diffractive grating surface including the one or more specific binding substances, and n is a total number of measurements collected.
x i and y i for i = 1, 2, 3, ...n, wherein x i is where a first measurement includes a first reflectance spectra of one or more specific binding substance attached to the colormetric resonant diffractive grating surface, y i a second measurement includes a second reflectance spectra of the one or more specific binding substances after a plurality of binding partners are applied to colormetric resonant diffractive grating surface including the one or more specific binding substances, and n is a total number of measurements collected.
79. The method of claim 75 wherein the step of determining a maximum value for a second measurement includes determining a maximum value y k such that:
(y k >= y i) for all i .noteq. k.
(y k >= y i) for all i .noteq. k.
80. The method of claim 75 wherein the step of determining whether the maximum value is greater than a pre-determined threshold includes:
computing a mean of the set of resonant reflectance data;
computing a standard deviation of the set of resonant reflectance data; and determining whether ((y k - mean)/standard deviation) is greater than a pre-determined threshold.
computing a mean of the set of resonant reflectance data;
computing a standard deviation of the set of resonant reflectance data; and determining whether ((y k - mean)/standard deviation) is greater than a pre-determined threshold.
81. The method of claim 75 wherein the step of defining a curve-fit region around the determined maximum value includes:
defining a curve-fit region of (2w+1) bins, wherein w is a pre-determined accuracy value;
extracting (x i, k -w <= i <= k + w); and extracting (y i, k - w <= i <= k + w).
defining a curve-fit region of (2w+1) bins, wherein w is a pre-determined accuracy value;
extracting (x i, k -w <= i <= k + w); and extracting (y i, k - w <= i <= k + w).
82. The method of claim 75 wherein the step of performing a curve-fitting procedure includes:
computing g i = 1n y i;
performing a 2nd order polynomial fit on g i to obtain g'i defined on (x i, k -w <= i <= k + w);
determining from the 2nd order polynomial fit coefficients a, b and c of for (ax2 + bx +c)-; and computing y'i = e g'i.
computing g i = 1n y i;
performing a 2nd order polynomial fit on g i to obtain g'i defined on (x i, k -w <= i <= k + w);
determining from the 2nd order polynomial fit coefficients a, b and c of for (ax2 + bx +c)-; and computing y'i = e g'i.
83. The method of claim 75 wherein the step of determining a location of a maximum resonant peak on the fitted curve includes:
determining location of maximum resonant peak (x p = (-b)/2a).
determining location of maximum resonant peak (x p = (-b)/2a).
84. The method of claim 75, wherein the step of determining a value of the maximum resonant peak includes determining the value with of x p at y'p.
85. A biosensor comprising a two-dimensional grating having a pattern of concentric rings, wherein the difference between an inside diameter and an outside diameter of each concentric ring is equal to about one-half of a grating period, wherein each successive ring has an inside diameter that is about one grating period greater than an inside diameter of a previous ring wherein when the structure is illuminated with an illuminating light beam, a reflected radiation spectrum is produced that is independent of an illumination polarization angle of the illuminating light beam, and wherein one or more specific binding substances are immobilized on the two-dimensional grating.
86. The biosensor of claim 85, wherein when the structure is illuminated a resonant grating effect is produced on the reflected radiation spectrum, wherein the depth and period of the two-dimensional grating are less than the wavelength of the resonant grating effect, and wherein a narrow band of optical wavelengths is reflected from the structure when the structure is illuminated with a broadband of optical wavelengths.
87. The biosensor of claim 85, wherein the two-dimensional grating has a period of about 0.01 microns to about 1 micron and a depth of about 0.01 microns to about 1 micron.
88. A biosensor comprising an array of holes or posts arranged such that the holes or posts are centered on corners and in the center of hexagons, wherein the hexagons are arranged in a closely packed array, wherein when the structure is illuminated with an illuminating light beam, a reflected radiation spectrum is produced that is independent of an illumination polarization angle of the illuminating light beam, and wherein one or more specific binding substances are immobilized on the array of holes or posts.
89. The biosensor of claim 88, wherein when the structure is illuminated a resonant grating effect is produced on the reflected radiation spectrum, wherein the depth or height and period of the holes or posts are less than the wavelength of the resonant grating effect, and wherein a narrow band of optical wavelengths is reflected from the structure when the structure is illuminated with a broad band of optical wavelengths.
90. The structure of claim 88, wherein the array holes or posts have a period of about 0.01 microns to about 1 micron and a depth of height of about 0.01 microns to about 1 micron.
91. A biosensor comprising:
(a) a first two-dimensional grating comprising a high refractive index material and having a top surface and a bottom surface;
(b) a second two-dimensional grating comprising a high refractive index material and having a top surface and a bottom surface, wherein the top surface of the second two-dimensional grating is attached to the bottom surface of the first two-dimensional grating; and (c) one or more specific binding substances or one or more specific binding substances bound to their binding partners immobilized on the top surface of the first two-dimensional grating;
wherein, when the biosensor is illuminated two resonant grating effects are produced on the reflected radiation spectrum, and wherein the depth and period of both of the two-dimensional gratings are less than the wavelength of the resonant grating effects.
(a) a first two-dimensional grating comprising a high refractive index material and having a top surface and a bottom surface;
(b) a second two-dimensional grating comprising a high refractive index material and having a top surface and a bottom surface, wherein the top surface of the second two-dimensional grating is attached to the bottom surface of the first two-dimensional grating; and (c) one or more specific binding substances or one or more specific binding substances bound to their binding partners immobilized on the top surface of the first two-dimensional grating;
wherein, when the biosensor is illuminated two resonant grating effects are produced on the reflected radiation spectrum, and wherein the depth and period of both of the two-dimensional gratings are less than the wavelength of the resonant grating effects.
92. The biosensor of claim 91, wherein a substrate layer supports the bottom surface of the second two-dimensional grating.
93. The biosensor of claim 91, further comprising a cover layer on the top surface of the first two-dimensional grating, wherein the one or more specific binding substances are immobilized on the surface of the cover layer opposite of the two-dimensional grating.
94. The biosensor of claim 91, wherein the top surface of the first two-dimensional grating is in physical contact with a test sample, and the second two dimensional grating is not in physical contact with the test sample.
95. The biosensor of claim 91, wherein when a peak resonant reflection wavelength is measured for the first and second two-dimensional gratings, the difference between the two measurements indicates the amount of one or more specific binding substances, binding partners, or both deposited on the surface of the first two-dimensional grating.
96. A biosensor comprising:
(a) a first two-dimensional grating comprising a high refractive index material and having a top surface and a bottom surface;
(b) a substrate layer comprising a top surface and a bottom surface, wherein the top surface of the substrate supports the bottom surface of the first two-dimensional grating;
(c) a second two-dimensional grating comprising a high refractive index material and having a top surface and a bottom surface, wherein the bottom surface of the second two-dimensional grating is attached to the bottom surface of the substrate;
and (d) one or more specific binding substances or one or more specific binding substances bound to their binding partners immobilized on the top surface of the first two-dimensional grating;
wherein, when the biosensor is illuminated two resonant grating effects are produced on the deflected radiation spectrum, and wherein the depth and period of both of the two-dimensional gratings are less than the wavelength of the resonant grating effects.
(a) a first two-dimensional grating comprising a high refractive index material and having a top surface and a bottom surface;
(b) a substrate layer comprising a top surface and a bottom surface, wherein the top surface of the substrate supports the bottom surface of the first two-dimensional grating;
(c) a second two-dimensional grating comprising a high refractive index material and having a top surface and a bottom surface, wherein the bottom surface of the second two-dimensional grating is attached to the bottom surface of the substrate;
and (d) one or more specific binding substances or one or more specific binding substances bound to their binding partners immobilized on the top surface of the first two-dimensional grating;
wherein, when the biosensor is illuminated two resonant grating effects are produced on the deflected radiation spectrum, and wherein the depth and period of both of the two-dimensional gratings are less than the wavelength of the resonant grating effects.
97. The biosensor of claim 96, further comprising a cover layer on the top surface of the first two-dimensional grating, wherein the one or more specific binding substances are immobilized on the surface of the cover layer opposite of the two-dimensional grating.
98. The biosensor of claim 96, wherein the top surface of the first two-dimensional grating is in physical contact with a test sample, and the second two dimensional grating is not in physical contact with the test sample.
99. The biosensor of claim 98, wherein when a peak resonant reflection wavelength is measured for the first and second two-dimensional gratings, the difference between the two measurements indicates the amount of one or more specific binding substances, binding partners, or both deposited on the surface of the first two-dimensional grating.
100. The biosensor of claim 1, further comprising an antireflective dielectric coating on a surface of the substrate opposite of the two-dimensional grating.
101. The biosensor of claim 1, wherein the biosensor is attached to a bottomless microtiter plate by a method selected from the group consisting of adhesive attachment, ultrasonic welding and laser welding.
102. A method of detecting an interaction of a first molecule with a second test molecule comprising:
(a) applying a mixture of the first and second molecules to a distinct location on a biosensor, wherein the biosensor comprises a two-dimensional grating comprising of a high refractive index material, and a substrate layer that supports the two-dimensional grating; and wherein, when the biosensor is illuminated a resonant grating effect is produced on the reflected radiation spectrum, and wherein the depth and period of the two-dimensional grating are less than the wavelength of the resonant grating effect;
(b) applying a mixture of the first molecule with a third control molecule to a distinct location on the biosensor of (a) or a similar biosensor, wherein the third control molecule does not interact with the first molecule, and wherein the third control molecule is about the same size as the first molecule; and (c) detecting a shift in the reflected wavelength of light from the distinct locations of step (a) and step (b);
wherein, if the shift in the reflected wavelength of light from the distinct location of step (a) is greater than the shift in the reflected wavelength in step (b), then the first molecule and the second test molecule interact.
(a) applying a mixture of the first and second molecules to a distinct location on a biosensor, wherein the biosensor comprises a two-dimensional grating comprising of a high refractive index material, and a substrate layer that supports the two-dimensional grating; and wherein, when the biosensor is illuminated a resonant grating effect is produced on the reflected radiation spectrum, and wherein the depth and period of the two-dimensional grating are less than the wavelength of the resonant grating effect;
(b) applying a mixture of the first molecule with a third control molecule to a distinct location on the biosensor of (a) or a similar biosensor, wherein the third control molecule does not interact with the first molecule, and wherein the third control molecule is about the same size as the first molecule; and (c) detecting a shift in the reflected wavelength of light from the distinct locations of step (a) and step (b);
wherein, if the shift in the reflected wavelength of light from the distinct location of step (a) is greater than the shift in the reflected wavelength in step (b), then the first molecule and the second test molecule interact.
103. The method of claim 102, wherein the first molecule is selected from the group consisting of a nucleic acid, polypeptide, antigen, polyclonal antibody, monoclonal antibody, single chain antibody (scFv), F(ab) fragment, F(ab')2 fragment, Fv fragment, small organic molecule, cell, virus, and bacteria.
104. The method of claim 102, wherein the second test molecule is selected from the group consisting of a nucleic acid, polypeptide, antigen, polyclonal antibody, monoclonal antibody, single chain antibody (scFv), F(ab) fragment, F(ab')2 fragment, Fv fragment, small organic molecule, cell, virus, and bacteria.
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- 2001-08-15 US US09/930,352 patent/US7094595B2/en not_active Expired - Lifetime
- 2001-08-15 US US09/929,957 patent/US7118710B2/en not_active Expired - Fee Related
- 2001-10-23 WO PCT/US2001/045455 patent/WO2002061429A2/en active Application Filing
- 2001-10-23 US US10/415,037 patent/US7435385B2/en active Active
- 2001-10-23 JP JP2002561944A patent/JP4202140B2/en not_active Expired - Fee Related
- 2001-10-23 EP EP01998024A patent/EP1337847A1/en not_active Withdrawn
- 2001-10-23 AU AU2002249786A patent/AU2002249786B2/en not_active Ceased
- 2001-10-23 AU AU2002246885A patent/AU2002246885A1/en not_active Abandoned
- 2001-10-23 CA CA2427689A patent/CA2427689C/en not_active Expired - Fee Related
- 2001-10-23 EP EP10185633A patent/EP2402739A1/en not_active Withdrawn
- 2001-10-23 WO PCT/US2001/050723 patent/WO2002059602A2/en active Application Filing
-
2004
- 2004-11-12 US US10/988,414 patent/US7422891B2/en not_active Expired - Lifetime
-
2005
- 2005-08-29 US US11/214,396 patent/US8101423B2/en not_active Expired - Fee Related
-
2008
- 2008-08-14 JP JP2008208966A patent/JP2009025312A/en active Pending
- 2008-08-14 JP JP2008208969A patent/JP2008309799A/en active Pending
- 2008-12-02 US US12/326,237 patent/US20090148955A1/en not_active Abandoned
Also Published As
Publication number | Publication date |
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AU2002249786B2 (en) | 2006-03-30 |
US7422891B2 (en) | 2008-09-09 |
AU2002246885A1 (en) | 2002-08-06 |
JP2009025312A (en) | 2009-02-05 |
WO2002061429A2 (en) | 2002-08-08 |
WO2002061429A8 (en) | 2003-05-15 |
EP1337847A1 (en) | 2003-08-27 |
WO2002059602A2 (en) | 2002-08-01 |
US8101423B2 (en) | 2012-01-24 |
WO2002059602A8 (en) | 2003-03-20 |
US7435385B2 (en) | 2008-10-14 |
US7094595B2 (en) | 2006-08-22 |
JP4202140B2 (en) | 2008-12-24 |
WO2002059602A3 (en) | 2003-01-30 |
CA2427689C (en) | 2012-01-24 |
US7118710B2 (en) | 2006-10-10 |
US20020168295A1 (en) | 2002-11-14 |
US20060030033A1 (en) | 2006-02-09 |
US20020127565A1 (en) | 2002-09-12 |
EP2402739A1 (en) | 2012-01-04 |
US20050227374A1 (en) | 2005-10-13 |
JP2004529324A (en) | 2004-09-24 |
US20040132172A1 (en) | 2004-07-08 |
JP2008309799A (en) | 2008-12-25 |
US20090148955A1 (en) | 2009-06-11 |
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