Photonic biosensor arrays

• US 8,389,299 B2
• Filed: 03/19/2008
• Issued: 03/05/2013
• Est. Priority Date: 03/23/2007
• Status: Active Grant

• Current Assignee: Attomarker Limited
• Original Assignee: Attomarker Limited
• Inventors: Shaw, Andrew Mark, Olkhov, Rouslan Vladamir, Barnes, William Leslie
• Primary Examiner(s): White, Dennis M
• Application Number:
  US12/531,814
• Publication Number:
  US 20100167946A1
• Time in Patent Office: 1,812 Days
• Field of Search: 422/68.1, 422/82.05, 422/82.06, 422/82.08, 422/82.09, 422/82.11, 422/99, 422/40, 435/164, 435/165, 435/283.1, 435/287.1, 435/287.2, 435/4, 435/5, 435/7.2, 435/7.9, 436/149, 436/164, 436/172, 436/174, 436/518, 436/805, 436/809, 356/369, 506/9
• US Class Current: 436/809
• CPC Class Codes: B82Y 15/00 Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: G01N 21/211 Ellipsometry optical thickness measurement G01B11/06
• CPC Class Codes: G01N 21/253 for batch operation, i.e. multisample apparatus analytical automats G01N35/00
• CPC Class Codes: G01N 21/554 detecting the surface plasmon resonance of nanostructured metals, e.g. localised surface plasmon resonance
• CPC Class Codes: G01N 33/54313 the carrier being characterised by its particulate form
• CPC Class Codes: G01N 33/54373 involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
• CPC Class Codes: G01N 33/553 Metal or metal coated
• CPC Class Codes: Y10T 436/25 including sample preparation

Abstract

This invention relates to photonic biosensor arrays in particular employing plasmon resonance based sensing, and to methods and apparatus for reading such arrays. A biosensor array for plasmon resonance-based sensing of a plurality of different biological targets simultaneously, the array comprising a transparent substrate having a surface bearing a plurality of assay spots for plasmon resonance sensing, each of said assay spots comprising a discrete metallic island, a said metallic island comprising a plurality of metallic nanoparticles to which are attached functionalizing molecules for binding to a said biological target, different said islands bearing different said functionalizing molecules for binding to different ones of said biological targets, and wherein total internal reflection of light at said surface at a wavelength at or near a said plasmon resonance results in scattering of said light away from said surface, said scattering being modulated by said binding of said biological targets.

Claims

1. A biosensor array for Plasmon resonance-based sensing of a plurality of different biological targets simultaneously, the array comprising a transparent substrate having a surface bearing a plurality of assay spots for Plasmon resonance sensing, each of said assay spots comprising a discrete metallic island, said metallic island comprising a plurality of metallic nanoparticles to which are attached functionalizing molecules for binding to said biological target, different said islands being different said functionalizing molecules for binding to different ones of said biological targets, and wherein total internal reflection of light at said surface at a wavelength at or near said plasmon resonance results in scattering of said light away from said surface, said scattering being modulated by said binding of said biological targets;

wherein said metallic islands of said array are spaced apart, arranged at intervals of at least 0.1 mm on a non-conducting substrate, and

wherein within said metallic island, said plurality of metallic nanoparticles comprise a discontinuous layer of individual, multi-faceted nanoparticles.

2. The biosensor array as claimed in claim 1, wherein said metallic nanoparticles have at least one dimension of less than 30 nm, and comprise rod-like nanoparticles.

3. The biosensor array as claimed in claim 1, wherein said nanoparticles include nanoparticles forming an optical antenna for said light, wherein said optical antenna comprises an adjacent pair of nanoparticles having a generally rod-like or triangular shape, and having adjacent ends separated by a gap of less than 50 nm, said nanoparticles having physical lengths which are resonant for said plasmon resonance at substantially the same optical wavelength.

4. The biosensor array as claimed in claim 1, further comprising at least one control spot substantially lacking said functionalizing molecules, and wherein said control spot comprises an array of metallic elements configured to produce an interference or fringe pattern when said control spot is viewed.

5. The biosensor array as claimed in claim 1, wherein said transparent substrate is configured as a Dove prism, and includes an edge or grating optical coupling for coupling said light into an edge of said transparent substrate of said array to waveguide said light within a thickness of said substrate.

6. The biosensor array as claimed in claim 1, wherein said metallic nanoparticles comprise gold nanoparticles, and wherein said different functionalizing molecules are attached to said metallic nanoparticles using the same ligand.

7. A method of using the biosensor array of claim 1 for a plasmon resonance-based sensing of a plurality of different biological targets simultaneously, the method comprising:

coupling light of at least one wavelength into said biosensor array such that total internal reflection of said light at said surface generates an evanescent wave field which excites plasmons in said functionalized metallic nanoparticles and scatters said light;

flowing a liquid carrying a plurality of said biological targets for analysis over said array;

imaging said scattered light from said array to generate image data for said biological targets carried by said liquid; and

analyzing said image data to determine levels of said biological targets carried by said liquid.

8. The method as claimed in claim 7, wherein said imaging of said scattered light comprises imaging said array from a side opposite to a side of said array being said assay spots.

9. The method as claimed in claim 7, wherein said imaging is performed in real-time to follow binding kinetics of said biological targets carried by said liquid, and wherein said analyzing comprises determining time variations of binding of said different targets to said different functionalizing molecules;

and further comprising matching said time variations to one or more pathological conditions associated with said biological targets carried by said liquid.

10. The method as claimed in claim 7, wherein said light comprises light of at least two different wavelengths one to either side of said plasmon resonance, and wherein said analyzing includes forming a ratio of signals at said two different wavelengths in said image data.

11. The method as claimed in claim 7, wherein said analyzing comprises comparing a signal from one of said assay spots with a signal from another of said assay spots to compensate for non-specific binding interactions.

12. The method as claimed in claim 7, wherein said analyzing comprises controlling for a bulk refractive index of said liquid using a control spot substantially lacking said functionalizing molecules, wherein said control spot is configured to form a diffraction or interference pattern in said imaged scattered light, wherein said controlling uses said diffraction or interference pattern.

13. The method as claimed in claim 7, wherein said biosensor array has an open top onto which said liquid may be deposited, and one or more built-in semiconductor light sources configured to provide waveguided light within said transparent substrate of said array to excite said plasmons.

14. The method as claimed in claim 7, wherein said imaging comprises imaging dark-field scattered light through a back surface of said biosensor array.

15. The biosensor array as claimed in claim 1, further comprising an apparatus for plasmon resonance-based sensing of a plurality of different biological targets, the apparatus comprising:

means for coupling light of at least one wavelength into said biosensor array such that total internal reflection of said light at said surface generates an evanescent wave field which excites plasmons in said functionalized metallic nanoparticles and scatters said light;

means for flowing a liquid carrying a plurality of said biological targets for analysis over said array;

means for imaging said scattered light from said array to generate image data for said biological targets carried by said liquid; and

means for analyzing said image data to determine levels of said biological, targets carried by said liquid.

16. The biosensor array as claimed in claim 15, wherein said means for imaging comprises means to image dark-field scattered light through a back surface of said array.

17. The biosensor array as claimed in claim 1, further comprising an apparatus for reading the biosensor array for Plasmon resonance-based sensing for plasmon resonance-based sensing of a plurality of different biological targets, the array comprising a transparent substrate having a surface bearing a plurality of assay spots for plasmon resonance sensing, each of said assay spots comprising a discrete metallic island to which is attached functionalizing molecules for binding to said biological target, different said islands bearing different said functionalizing molecules for binding to different ones of said biological targets, and wherein total internal reflection of light at said surface at a wavelength at or near said plasmon resonance results in scattering of said light away from said surface, said scattering being modulated by said binding of said biological targets, the apparatus comprising:

a light source to generate light at two different substantially monochromatic wavelengths;

an optical coupling device to couple light from said light source into said sensing array;

an imaging system to image light from said array at said two different wavelengths scattered by plasmon resonance in said metallic islands, and to generate image date; and

an image analysis system coupled to said imaging system to receive said image data and to analyze said plasmon-resonance scattered light to determine levels of said different biological targets attached to said array.

18. The apparatus as claimed in claim 17, configured to determine said levels of said different biological targets attached to said array using light of two orthogonal polarizations.

19. A method of using the biosensor array of claim 1 for plasmon resonance-based sensing of different biological targets simultaneously, the method comprising:

coupling light of at least one wavelength into said biosensor array such that total internal reflection of said light at said surface generates am evanescent wave field which excites plasmons in said functionalized metallic nanoparticles, said light comprising polarized light with polarization modulation;

detecting an orientation of an elliptical of totally internally reflected modulated polarized light; and

outputting a target sensing signal responsive to binding of said biological target to said functionalizing molecule dependent on detection of a change in said elliptical polarization orientation, wherein s- and p-polarization components of said polarized light do not self-interfere.

20. An apparatus for reading a plasmon resonance sensing array for plasmon resonance-based sensing of a plurality of different biological targets, the array comprising a transparent substrate having a surface being a plurality of assay spots for said plasmon resonance testing, the apparatus comprising:

a light source to generate polarized light;

a polarization modulator to modulate a polarization of said polarized light;

an optical system to illuminate said array with said modulated polarized light to generate plasmons in said assay spots by total internal reflection of said polarized light wherein s- and p-polarization components of said polarized light do not self-interfere;

a detection system to detect an orientation of an elliptical polarisation of said totally internally reflected modulated polarized light; and

an output to output a target sensing signal dependent on binding of said biological target to said functionalizing molecule responsive to detection of a change in said elliptical polarization orientation;

and said plasmon resonance sensing array wherein

each of said assay spots comprising a discrete metallic island, said metallic island comprising a plurality of metallic nanoparticles to which are attached functionalizing molecules for binding to said biological target, different said islands bearing different said functionalizing molecules for binding to different ones of said biological targets;

wherein said metallic islands of said array are spaced apart, arranged at intervals of at least 0.1 mm on a non-conducting substrate and

wherein within said metallic island, said plurality of metallic nanoparticles comprise a discontinuous layer of individual, multi-faceted nanoparticles.

21. The apparatus as claimed in claim 20, wherein said polarization modulator is pixelated for separate interrogation of said assay spots, and wherein said detection system is configured to enable separation of signals from different ones of said assay spots.

22. The apparatus as claimed in claim 20, configured to image dark-field scattered light through a back surface of said array.