Microarray scanning
Methods and apparatus for scanning of a microarray to provide an image of the microarray are disclosed. Data related to change in polarization state of a scanning light beam for individual points or lines or other portion of the microarray are collected and processed to image the microarray at high resolution and speed.
1. An apparatus for scanning, comprising:
a light source emitting a polarized light beam;
an optical assembly having a surface adapted to allow placing thereon a specimen array, the light beam from the light source being reflected by the surface to provide an evanescent field over a portion of the specimen array such that the portion of the specimen array in the evanescent field causes a polarization change in the light beam; and
a detector positioned to detect the polarization change in the light beam as the light beam is scanned over the specimen array.
2. The apparatus of claim 1 , wherein the light source comprises a quasi-monochromatic light source of moderate bandwidth.
3. The apparatus of claim 2 , wherein the quasi-monochromatic light source of moderate bandwidth is a light-emitting diode (LED) or a superluminescent diode (SLD).
4. The apparatus of claim 2 , wherein the quasi-monochromatic light source of moderate bandwidth comprises an incandescent source and an optical filter, the light emitted from the incandescent source passing through the optical filter, the optical filter limiting the wavelengths of the light transmitted through the optical filter such as to constitute quasi-monochromatic light of moderate bandwidth.
5. The apparatus of claim 1 , wherein the light source comprises a laser emitting substantially coherent light, and further comprising an optical diffuser mechanically attached to a mechanical actuator, the light emitted from the laser passing through the diffuser, the diffuser being moved with respect to the laser by the actuator, the movement of the diffuser with respect to the laser creating fluctuations in the speckle pattern of light detected by the detector, the fluctuations being adapted to remove speckle effects from the light detected by the detector.
6. The apparatus as in claim 5 , wherein the mechanical actuator is a motor rotating the optical diffuser.
7. The apparatus of claim 1 , wherein the light source comprises a beam forming system, the beam forming system causing the light emerging from the light source to be collimated.
8. The apparatus of claim 1 , wherein the light source comprises an optical polarizer.
9. The apparatus of claim 8 , wherein the optical polarizer is controllably rotated by a motor.
10. The apparatus of claim 1 , wherein the light source comprises an optical retarder, the retarder introducing an optical phase shift between two orthogonal components of light passing through the retarder.
11. The apparatus of claim 10 , wherein the optical retarder is controllably rotated by a motor.
12. The apparatus of claim 10 , wherein the optical retarder changes retardance according to an externally introduced physical parameter.
13. The apparatus of claim 1 , wherein the optical assembly comprises an optical prism.
14. The apparatus of claim 1 , wherein the specimen array comprises a two-dimensional array formed of multiple fields comprising biomolecular substances.
15. The apparatus of claim 14 , wherein the biomolecular substances are proteins, peptides, and/or polynucleotide sequences.
16. The apparatus of claim 1 , wherein the detector comprises a two-dimensional array detector.
17. The apparatus of claim 16 , wherein the detector comprises an optical polarizer, a two-dimensional CCD array, and/or a two-dimensional photodiode array.
18. The apparatus of claim 1 , wherein the light beam is in the form of a line.
19. The apparatus of claim 1 , further comprising a translation stage operably coupled to the optical assembly.
20. The apparatus of claim 1 , further comprising an actuator operably coupled to the light source.
21. The apparatus of claim 1 , further comprising an actuator operably coupled to the detector.
22. The apparatus of claim 1 , further comprising a processor coupled to the detector, the processor processing the signal from the detector to obtain a representation of the optical phase shift occurring in the portion of the specimen array.
23. The apparatus of claim 1 , further comprising a processor coupled to the detector, the processor processing a plurality of signals related to spatially distributed polarization changes as the light beam scans the specimen array to thereby provide an image of the specimen array.
24. The apparatus of claim 23 , wherein the processor provides feedback to reduce or increase resolution over a portion of the microarray.
25. The apparatus of claim 23 , wherein the processor provides feedback to skip scanning over a portion of the microarray.
26. The apparatus of claim 23 , wherein the processor provides feedback to increase the frequency of scanning over a portion of the microarray.
27. An apparatus for scanning, comprising:
a light source emitting a polarized light beam;
an optical assembly having a surface adapted to allow placing thereon a specimen array, the light beam from the light source being reflected by the surface to provide an evanescent field over a portion of the specimen array such that the portion of the specimen array in the evanescent field causes a polarization change in the light beam;
a detector positioned to detect the polarization change in the light beam as the light beam is scanned over the specimen array; and
a processor to process a plurality of signals related to spatially distributed polarization changes to thereby provide feedback for subsequent scanning of the specimen array.
28. The apparatus of claim 27 , wherein the processor provides feedback to reduce or increase resolution over a portion of the microarray.
29. The apparatus of claim 27 , wherein the processor provides feedback to skip scanning over a portion of the microarray.
30. The apparatus of claim 27 , wherein the processor provides feedback to decrease or increase the frequency of scanning over a portion of the microarray.
31. A method of imaging, comprising:
passing a polarized light beam into an optical structure for reflection at a surface of the optical structure to provide an evanescent field over a portion of a specimen array, the portion of the specimen array in the evanescent field causing a polarization change in the light beam;
detecting the polarization change in the light beam as the light beam scans the specimen array; and
processing a plurality of signals related to spatially distributed polarization changes to thereby provide an image of the specimen array.
32. The method of claim 31 , further comprising using the plurality of spatially distributed polarization changes to determine two-dimensionally distributed presence and/or properties of the specimen array constituents.
33. The method of claim 31 , wherein the specimen array is in a micro-titer plate.
34. The method of claim 33 , further comprising:
resolving the plurality of spatially distributed polarization changes for matching positions in the micro-titer plate; and
analyzing the polarization changes to determine desired characteristics in each position.
35. The method of claim 31 , wherein the specimen array is a series of discrete specimen spots.
36. The method of claim 35 , further comprising analyzing the polarization changes to determine the binding characteristics of each discrete specimen spot.
37. The method of claim 31 , wherein a specimen array having no molecular tagging is placed in the evanescent field.
38. A method of scanning, comprising:
passing a polarized light beam into an optical structure for reflection at a surface of the optical structure to provide an evanescent field over a portion of a specimen array, the portion of the specimen array in the evanescent field causing a polarization change in the light beam;
detecting the polarization change in the light beam as the light beam scans the specimen array; and
processing a plurality of signals related to spatially distributed polarization changes to thereby provide feedback for subsequent scanning of the specimen array.
39. The method of claim 38 , wherein the subsequent scanning decreases or increases resolution over a portion of the microarray.
40. The method of claim 38 , wherein the subsequent scanning skips over a portion of the microarray.
41. The method of claim 38 , wherein the subsequent scanning decreases or increases the frequency of scanning over a portion of the microarray.