Biological testing with sawtooth-shaped prisms
In one embodiment, an apparatus is provided for testing for the presence of analytes in a sample. The apparatus comprises a source of light directed at the bottom surface of a substrate to achieve total internal reflection and to generate an evanescent field. An array of capture elements is immobilized on the top surface of the substrate. The bottom surface of the substrate is configured as a sawtooth (in cross section), the “teeth” aligned with the rows or the columns in the array. The outer faces of the sawtooth “prisms” are non-parallel to the substrate top surface, and specific requirements are imposed on the prism light-entrance face, and the substrate thickness and refractive index.
1. An apparatus for determining the presence of analytes in a sample, the apparatus comprising:
a transparent substrate with top and bottom surfaces spaced apart a distance d and formed of a material having an index of refraction n, wherein the substrate bottom surface includes a sawtooth prism having a cross-sectional sawtooth configuration comprising a light input portion and a light output portion which are mirror images of each other about a center line of the sample, and wherein the sawtooth prism has a height H, a pitch P, and an angle θ between an entry face and the plane of the substrate top surface according to the equation H=P sin θ cos θ.
2. The apparatus as in claim 1 , wherein the substrate comprises a microscope slide.
3. The apparatus as in claim 1 , further comprising an array of capture elements immobilized on the substrate top surface.
4. The apparatus as in claim 3 , further comprising a plurality of arrays in spaced apart positions on the top surface of the substrate.
5. The apparatus as in claim 3 , further comprising first and second coatings relating to different sensitivities to molecular attachment and wherein the array partially overlaps each of the coatings.
6. The apparatus as in claim 3 , further comprising a fluid channel in communication with the array and a sample input port connected to the channel, the channel being dimensioned to move a sample by capillary action across the array.
7. The apparatus as in claim 1 , further comprising a well-forming member integral with the substrate top surface, the member having a plurality of holes therein for forming sample wells, each of the wells in registry with an array.
8. The apparatus as in claim 5 , further comprising a well forming member integral with the substrate top surface, the member having a plurality of holes therein for forming sample wells, each of the wells in registry with the array.
9. The apparatus as in claim 7 , further comprising an optical system for directing light at an input face of at least one tooth of the sawtooth prism in a manner to obtain total internal reflection and an evanescent field in the plane of the top surface.
10. The apparatus as in claim 9 , further comprising an imaging system for imaging the pattern of light reflected from the array and for determining patterns of changes from an initial stored light pattern.
11. The apparatus as in claim 1 , wherein P >40 microns.
12. The apparatus as in claim 1 , wherein the cross-sectional sawtooth configuration further comprises in each tooth two sides of unequal lengths, and wherein X=P cos θ where X is the length of the shorter one of the two sides of the cross-sectional sawtooth configuration.
13. The apparatus as in claim 12 , wherein the substrate comprises glass at least 1 mm thick and the sawtooth prism comprises a film of two layers, including a COC planar film and a UV-cured acrylic ester.
14. The apparatus as in claim 5 , wherein the first and second coatings are taken from a group consisting of silicon dioxide, magnesium fluoride, tantalum pentoxide, titanium dioxide, and other metal oxides.
15. The apparatus as in claim 1 , wherein the sawtooth prism comprises a separate film attached to the substrate bottom surface.
16. An apparatus comprising:
a substrate having a top surface for placement of a sample and a bottom surface, wherein the bottom surface has a consecutive tooth-shaped geometry with teeth arranged in rows and columns, with consecutive teeth in a column abutting one another to form a sawtooth configuration comprising a light input portion and a light output portion which are minor images of each other about a center line of the sample and with adjacent teeth in a row being in alignment with one another.
17. The apparatus as in claim 16 , further comprising a well-defining member with an array of apertures there through attached to the top surface of the substrate to define an array of wells.
18. The apparatus as in claim 17 , wherein adjacent teeth in a row form an elongated wedge.
19. The apparatus as in claim 18 , wherein consecutive wedges comprise adjacent faces non-parallel to the plane of the top surface and form a continuous sawtooth configuration.
20. The apparatus as in claim 16 , wherein the bottom surface has a sawtooth geometry formed by an attached thin film with an embossed or preformed sawtooth geometry.
21. The apparatus as in claim 16 , further comprising a source of light of wavelength λ directed at an entrance face of at least one tooth of a sawtooth geometry wherein the area of the entrance face, X, is at least
λ
sin
φ
where φ is a divergence half-angle of the incident beam.
22. The apparatus as in claim 21 , wherein a divergence of light due to diffraction in the substrate is less than or equal to 0.5°, and the pitch P from tooth to tooth is P ≧100μm sin θ tan θ, where θ is the angle of incidence of the incoming light with respect to the plane of the top surface.
23. The apparatus as in claim 17 , further comprising an array of capture elements in each well.
24. The apparatus as in claim 17 , further comprising first and second coatings in each well wherein the coatings are selected to correspond to different levels of molecular attachment.
25. The apparatus as in claim 24 , wherein the capture elements in each well partially overlap each of the first and second coatings corresponding to different levels of molecular attachment.