Optical device for surface-generated fluorescence

• US 20050051733A1
• Filed: 06/03/2004
• Published: 03/10/2005
• Est. Priority Date: 06/06/2003
• Status: Active Grant

• Current Assignee: Oerlikon Surface Solutions Ag, PfAffikon (OC Oerlikon Corporation AG)
• Original Assignee: Unaxis Balzers (Unaxis)
• Inventors: Wiki, Max, Edlinger, Johannes
• Granted Patent: US 7,285,789 B2
• Time in Patent Office: Days
• US Class Current: 250/453.110
• CPC Class Codes: G01N 2021/6471 Special filters, filter wheel
• 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 21/6452 Individual samples arranged in a regular 2D-array, e.g. multiwell plates

Abstract

A sample substrate adapted for use with electromagnetic excitation light includes a base and a layer system. The layer system includes a multilayer interference coating with at least two layers wherein the thicknesses of the layers ensure that light emitted by a fluorescent sample material disposed on top of said multilayer interference coating is reflected. Light directed to a fluorescent sample material disposed on the substrate causes light to be emitted from the sample. The layer system includes a multilayer interference coating with at least two layers wherein thicknesses of the layers cause separation of the excitation light from the emitted light.

Claims

1. A sample substrate adapted for use with electromagnetic excitation light, comprising a base and a layer system, the layer system comprising a multilayer interference coating with at least two layers wherein the thicknesses of the layers ensure that light emitted by a fluorescent sample material disposed on top of said multilayer interference coating is reflected.

2. A sample substrate adapted for use with electromagnetic excitation light directed to a fluorescent sample material disposed on the substrate, causing light to be emitted from the sample, comprising a base and a layer system, the layer system comprising a multilayer interference coating with at least two layers wherein thicknesses of the layers cause separation of the excitation light from the emitted light.

3. The substrate according to claim 2 wherein the emitted light is adapted to be received by a photodetector on a front side of the fluorescent sample material opposite the substrate and the thicknesses of the layers cause reflection of substantially all of the emitted light.

4. The substrate according to claim 2 wherein the thicknesses of the layers are chosen in such a way that for s-polarized light impinging on the layer system and having a wavelength corresponding to a wavelength of the emitted light and a medium of incidence of the light having an index of refraction of 1.4, less than 1.5% of light is transmitted for angles of incidence from −

70 degrees to +70 degrees.

5. The substrate according to claim 3 wherein the thicknesses of the layers cause reflection of the emitted light in two different wavelength bands.

6. The substrate according to claim 3, wherein the thicknesses of the layers cause reflection of the emitted light in two different wavelength bands, wherein one wavelength band ranges from 550 nm to 600 nm and another wavelength band ranges from 650 nm to 700 nm.

7. The substrate according to claim 3, wherein the thicknesses of the layers cause reflection of the emitted light in two different wavelength bands and wherein the thicknesses of the layers cause transmission of not absorbed excitation light for two different excitation wavelengths.

8. The substrate according to claim 3, wherein the thicknesses of the layers cause reflection of the emitted light in two different wavelength bands, wherein one wavelength band ranges from 550 nm to 600 nm and another wavelength band ranges from 650 nm to 700 nm and wherein the thicknesses of the layers cause transmission of the excitation light for a first and a second excitation wavelength, wherein said first excitation wavelength is within the wavelength range of 532 nm to 548 nm and said second excitation wavelength is around 633 nm.

9. The substrate according to claim 3 where in the excitation light is waveguided within the layer system.

10. The substrate according to claim 9 wherein the emitted light is not waveguided within the layer system.

11. The substrate according to claim 3 wherein the excitation light is received at a front side of the fluoresecent sample material opposite the substrate and the thicknesses of the layers cause transmission of substantially all of the excitation light which is not absorbed by the fluorescent sample material.

12. The substrate according to claim 11 wherein greater than 90% of the excitation light which is not absorbed by the fluorescent sample material is transmitted at a 0 degree angle of incidence.

13. The substrate according to claim 11 wherein the thicknesses of the layers cause transmission of the excitation light in two different wavelength bands.

14. The substrate according to claim 2 wherein the excitation light is received at a front side of the fluorescent sample material opposite the substrate and the thicknesses of the layers cause reflection of substantially all of the excitation light.

15. The substrate according to claim 14 wherein the thicknesses of the layers cause reflection of the excitation light in two different wavelength bands.

16. The substrate according to claim 14 wherein the emitted light is adapted to be received by a photodetector on a rear side of the sample adjacent the substrate and the thicknesses of the layers cause transmission of substantially all of the emitted light.

17. The substrate according to claim 16 wherein the thicknesses of the layers cause transmission of the emitted light in two different wavelength bands.

18. The substrate according to claim 2 wherein the excitation light is received at a rear side of the fluorescent sample material adjacent the substrate and the thicknesses of the layers cause transmission of substantially all of the excitation light.

19. The substrate according to claim 18 wherein the thicknesses of the layers cause transmission of the excitation light in two different wavelength bands.

20. The substrate according to claim 18 wherein the emitted light is adapted to be received by a photodetector on a rear side of the sample adjacent the substrate and the thicknesses of the layers cause transmission of substantially all of the emitted light.

21. The substrate according to claim 20 wherein the thicknesses of the layers cause transmission of the emitted light in two different wavelength bands.

22. The substrate of claim 2 wherein the multilayer interference coating comprises dielectric layers having alternately relatively higher and lower refractive indices such that an outermost layer is one of the layers having a relatively lower refractive index.

23. The substrate of claim 22 wherein the layers comprise at least one of Nb2O5, SiO2, aluminum oxide, magnesium oxide;

oxides of the groups VIb, Vb, IVb, scandium, yttrium, calcium, strontium, zinc, iron, indium, tin, cerium, or holmium;

oxides of mixtures or alloys of scandium, yttrium, calcium, strontium, zinc, iron, indium, tin, cerium, or holmium; and

oxynitrides of Ti, Ta, Zr, Si, Hf, or Al;

fluorides of magnesium, barium, strontium, calcium, rare earths, and lead.

24. The substrate of claim 22 wherein the layers comprise Nb2O5 and SiO2.

25. The substrate of claim 2 further comprising a linking coating disposed on an outermost layer of the layer system.

26. The substrate of claim 25 wherein the linking coating layer is biologically active.

27. The substrate of claim 2 wherein the base is rigid.

28. The substrate of claim 2 wherein the base comprises one of glass, plastic, metal, or semiconductor material.

31. Use of the substrate according to claim 2 in bioanalysis systems

32. A fluorescence imaging system comprising a sample substrate according to claim 2 and a light source directed to the sample material on said sample substrate, said light including the electromagnetic excitation wavelength and being particular to a specified electromagnetic constituent of said sample material

33. A laser scanner comprising an electromagnetic imaging system according to claim 32.

29. A method of manufacturing a sample substrate adapted to obtain electromagnetic emission from a sample disposed on the substrate when the sample is excited by a light comprising the steps of:

selecting at least two layers of a reflecting multilayer interference coating;

determining thicknesses of the layers to cause separation of the excitation light from the emitted light; and

depositing the layers on a base.

30. The method of claim 29 further comprising the step of depositing a linking coating on the first layer system.