Grating-based sensor combining label-free binding detection and fluorescence amplification and readout system for sensor

• US 20100003743A1
• Filed: 04/30/2009
• Published: 01/07/2010
• Est. Priority Date: 08/11/2005
• Status: Active Grant

• Current Assignee: X-Body Incorporated (Bristol-Myers Squibb Company)
• Original Assignee: Sru Biosystems Incorporated
• Inventors: Borsody, Alex, Li, Peter Y., Binder, Brant, Jogikalmath, Gangadhar, Cunningham, Brian T., Laing, Lance G., Schulz, Stephen C.
• Granted Patent: US 7,960,170 B2
• Time in Patent Office: Days
• US Class Current: 435/288.7
• CPC Class Codes: G01N 21/552 Attenuated total reflection
• CPC Class Codes: G01N 21/6428 Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 21/648 using evanescent coupling or surface plasmon coupling for the excitation of fluorescence
• CPC Class Codes: G01N 21/7743 the reagent-coated grating coupling light in or out of the waveguide
• CPC Class Codes: Y10S 435/808 Optical sensing apparatus
• CPC Class Codes: Y10S 436/805 Optical property
• CPC Class Codes: Y10T 436/143333 Saccharide [e.g., DNA, etc.]

Abstract

A grating-based sensor is disclosed that has a grating structure constructed and designed for both evanescent resonance (ER) fluorescence detection and label-free detection applications. Some embodiments are disclosed which are optimized for ER detection in an air mode, in which the sample is dry. Other embodiments are optimized for ER detection in liquid mode, in which the sample is suspended in liquid medium such as water. One and two-dimensional gratings are also disclosed, including gratings characterized by unit cells with central posts, central holes, and two-level, two-dimensional gratings. A readout system for such sensors is also disclosed. One embodiment includes a first light source optimized for collecting label-free detection data, a second light source optimized for collecting ER fluorescence amplification data, and at least one detector. In one embodiment, the detector is an imaging system and includes a CCD camera for collecting both ER and label-free data. In other embodiments, the at least one detector takes the form of a spectrometer for collection of label-free data and a photomultiplier for collecting ER data. In other embodiments, a single light source such as a tunable laser or broad band light source is used.

Claims

1. A sensor for testing a sample placed on the sensor, comprising:

a substrate having a periodic surface grating structure wherein the periodic grating structure is constructed in a manner designed for both

     1) optical interrogation of the sensor with light in an evanescent resonance (ER) detection mode, and

     2) optical interrogation of the sensor with light in a label-free detection mode.

2. The sensor of claim 1, wherein the sample is in an air medium, and further comprising a detection instrument which includes at least one light source for optical interrogation of the sensor in the ER and label-free detection modes, and wherein the light from the at least one light source has a polarization perpendicular to the grating structure.

3. The sensor of claim 1, wherein the sample is in a liquid medium, and further comprising a detection instrument which includes at least one light source for optical interrogation of the sensor and wherein the light from the at least one light source has a polarization parallel to the grating structure.

4. The sensor of claim 2, wherein the light from the at least one light source has a wavelength selected to activate a luminescent material bound to (a) the sample, (b) the surface of the sensor, or (c) both (a) and (b).

5. The sensor of claim 2, wherein the light from the at least one light source interrogates the sensor at or near normal incidence.

6. The sensor of claim 2, wherein the detection instrument includes a software module calculating a peak wavelength value of reflected light for the sample in a label-free detection mode.

7. The sensor of claim 3, wherein the light from the at least one light source has a wavelength selected to activate a luminescent material bound to (a) the sample, (b) the surface of the sensor, or (c) both (a) and (b).

8. The sensor of claim 3, wherein the light from the at least one light source interrogates the sensor at or near normal incidence.

9. The sensor of claim 3, wherein the detection instrument includes a software module calculating a peak wavelength value of reflected light for the sample in a label-free detection mode.

10. The sensor of claim 1, wherein the grating structure comprises a two-dimensional periodic grating structure, the periodic grating structure is periodic in first and second dimensions, and wherein the first and second dimensions are mutually orthogonal.

11. The sensor of claim 10, wherein the first dimension of the periodic grating structure comprises a grating structure designed for label-free detection, the second dimension of the periodic grating structure comprises a grating structure designed for ER detection, and wherein the depth of the grating structure in the first dimension is substantially greater than the depth of the grating structure in the second dimension.

12. The sensor of claim 11, wherein the grating structure further comprises a laminate comprising a substrate, a layer having a grating structure bonded to the substrate, an SiO₂ layer deposited on the layer having the grating structure, and a layer of relatively high index of refraction material deposited on the SiO₂ layer.

13. The sensor of claim 11, wherein the grating structure in the first dimension has a period of between 260 and about 1500 nm and a depth of the grating between about 100 nm and about 3000 nm, and wherein the grating structure in the second dimension has a period of between about 200 nm and about 1000 nm, and wherein the depth of the grating in the second dimension is between about 10 nm and about 300 nm.

14. The sensor of claim 12, wherein the SiO₂ layer comprises a layer having a depth of between about 500 and about 5000 Angstroms.

15. The sensor of claim 12, further comprising a luminescent material bound to the layer of high index of refraction material.

16. The sensor of claim 11, wherein the grating structure comprises an array of unit cells having a two-level, two-dimensional form.

17. The sensor of claim 1, wherein the grating structure has a grating depth to half period ratio of between 0.6 and 1.2.

18. The sensor of claim 1, wherein the grating structure comprise array of unit cells each having a two-level, two-dimensional form comprising a first periodic grating structure of alternating high and low regions extending in a first dimension constructed in a manner designed for label-free detection and having a second periodic grating structure having alternating high and low regions superimposed on the first periodic grating structure extending in a second dimension orthogonal to the first dimension constructed in a manner designed for ER detection mode.

19. The sensor of claim 10, wherein the grating structure comprises an array of two dimensional unit cells each comprising a hole in the surface of the biosensor.

20. The sensor of claim 10, wherein the grating structure comprises an array of f unit cells each comprising a post projecting from the surface of the biosensor.

21. The sensor of claim 10, wherein the grating structure comprises a checkerboard configuration with alternating high and low regions.

22. The sensor of claim 10, wherein the grating structure in the first and second orthogonal dimensions has a different periodicity in the first and second dimensions, the periodicity in the first dimension is constructed so as to provide a broad resonance for incident light from the first light source at or near normal incidence with a wavelength tuned to excite a fluorophore associated with the sample, and the periodicity in the second dimension is constructed so as to yield a sharp resonance for illumination with light from the second light source in the near infra-red portion of the spectrum.

23. The sensor of claim 1, further comprising at least one sample placed on the biosensor, wherein the sample is selected from the group of samples consisting of molecules having a molecular weight of less than 1000 daltons, molecules with a molecular weight of between 1000 and 10,000 daltons, amino acids, proteins, nucleic acids, lipids, carbohydrates, nucleic acid polymers, viral particles, viral components, cellular components, and extracts of viral or cellular components, 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, polymers, peptide solutions, protein solutions, chemical compound library solutions, single-stranded DNA solutions, double stranded DNA solutions, combinations of single and double stranded DNA solutions, RNA solutions and biological samples.

24. The sensor of claim 23, wherein the biological samples comprise samples selected from the group of 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 prostatic fluid.

25. The sensor of claim 1, further comprising a sample placed on the sensor, wherein the ER detection mode detects natural fluorescence of the sample.

26. The sensor of claim 1, further comprising a sample placed on the biosensor, wherein a fraction of the sample is bound to an inhibitor.

27. The sensor of claim 26, wherein the inhibitor is bound to a fluorescent label.

28. The sensor of claim 26, wherein the sample comprises a protein.