Scanning of biological samples

• US 6,867,851 B2
• Filed: 08/02/2002
• Issued: 03/15/2005
• Est. Priority Date: 11/04/1999
• Status: Expired due to Term

• Current Assignee: Regents of The University of Minnesota (University of Minnesota)
• Original Assignee: Regents of The University of Minnesota (University of Minnesota)
• Inventors: Nelson, Scott A., Lewis, Steven A., Anderson, Kraig, Talghader, Joseph J., Blumenfeld, Martin, Sanders, Mark A.
• Primary Examiner(s): Nguyen, Tu T.
• Application Number:
  US10/211,113
• Publication Number:
  US 20030151735A1
• Time in Patent Office: 956 Days
• Field of Search: 356/73, 356 36- 50, 356300-343, 250/458.1, 436164-169, 436805-811, 436175-180, 422102-106
• US Class Current: 356/73
• CPC Class Codes: G01N 2021/6484 Optical fibres
• 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
• CPC Class Codes: G01N 21/6456 Spatial resolved fluorescence measurements; Imaging
• CPC Class Codes: G02B 21/06 Means for illuminating specimens
• CPC Class Codes: G02B 21/16 adapted for ultraviolet illumination ; Fluorescence microscopes G02B21/0076 takes precedence

Abstract

An image detection apparatus comprises a light source and a holding apparatus adapted to hold a substrate on which a biological sample may be mounted. Light from the light source impinges on the biological sample and causes light that is representative of the sample to be emitted from the sample. A light detector is positioned in the path of the emitted light and is scanned across the emitted light. The emitted light may be focused on the light detector with an optical assembly. The emitted light may be generated by chromophores on probes in the biological sample.

Claims

1. An image detection apparatus for use with a biological sample, the apparatus comprising:

a light source that provides source light along a source light path;

a holding apparatus to hold a substrate, on which a biological sample is mounted, in a position such that at least a portion of the source light impinges on at least a portion of the biological sample and causes light emission representative of the biological sample to be produced in the form of emitted light along an emitted light path;

a light detector array comprising a plurality of detector pixels that is positioned in at least a portion of the emitted light path such that the array senses emitted light from the biological sample; and

a transport mechanism adapted to translate the light detector array to allow the capture of an image of at least a portion of the biological sample as a plurality of sub-images.

2. The image detection apparatus of claim 1, wherein the light source comprises one or more light emitting diodes or laser diodes.

3. The image detection apparatus of claim 2, wherein the emitted light from a single micro-array address is directed to substantially one detector pixel.

4. The image detection apparatus of claim 1, wherein the biological sample is in the form of a micro-array comprising one or more sequences of nucleic acids immobilized to a substrate, each sequence immobilized at a particular micro-array address, wherein the nucleic acid sequences are positioned such that emitted light from a micro-array address is substantially directed onto at least one detector pixel of the light detector array.

5. The image detection apparatus of claim 1, wherein the substrate comprises an opaque material.

6. The image detection apparatus of claim 1, wherein the emitted light path does not pass through any part of the substrate.

7. An image detection system for use with one or more biological samples, the system comprising:

a light source that provides source light along a source light path;

a holding apparatus adapted to hold a substrate supporting a biological sample in at least a portion of the source light path such that the source light impinges on at least a portion of the biological sample, and produces emitted light representative of the biological sample along an emitted light path, wherein the source light path and the emitted light path are both on a first side of the substrate, and

a light detector positioned in at least a portion of the emitted light path, and scannable across an image that is representative of the biological sample.

8. The image detection system of claim 7, wherein the light source comprises a plurality of light emitting diodes or laser diodes.

9. The image detection system of claim 8, wherein the plurality of light emitting diodes comprise a first group of one or more diodes that emit light at a first wavelength range, and a second group of one or more diodes that emit light at a second wavelength range that is substantially different from the first wavelength range.

10. The image detection system of claim 7, further comprising a source light filter positioned in the source light path to reduce interference between the source light and the emitted light.

11. The image detection system of claim 7, further comprising an illumination optics assembly positioned in the source light path, and having a light pipe and a toroidal or semi-toroidal focusing lens.

12. The image detection system of claim 11, wherein the illumination optics assembly comprises an excitation filter.

13. The image detection apparatus of claim 7, further comprising a viewing optics assembly positioned in the emitted light path for focusing the emitted light on the light detector.

14. The image detection apparatus of claim 13, further comprising a fixed focus imaging system.

15. The image detection apparatus of claim 14, wherein the apparatus has a depth of field greater than about 50 microns.

16. The image detection apparatus of claim 13, further comprising an emission filter positioned in the emitted light path between the viewing optics assembly and the detector.

17. The image detection system of claim 7, wherein the substrate has a first surface and a second surface opposing the first surface, and the biological sample is in the form of a micro-array comprising one or more sequences of nucleic acids immobilized to the first surface of the substrate, each sequence immobilized at a particular micro-array address, wherein the nucleic acid sequences are positioned on the micro-array such that light emitted from one micro-array address is substantially directed onto one detector pixel of the light detector.

18. The image detection system of claim 17, wherein the biological sample comprises a probe and a light emitting or light absorbing label.

19. The image detection system of claim 17, wherein the light from a single micro-array address is directed to substantially one detector pixel.

20. The image detection system of claim 7, wherein the emitted light comprises light of fluorescence from the biological sample.

21. The image detection system of claim 7, further comprising a lens assembly positioned in the emitted light path to focus the emitted light on the light detector.

22. The image detection system of claim 7, wherein the light detector comprises a linear array that further comprises a plurality of detector pixels.

23. The image detection system of claim 22, wherein the light detector comprises a charge coupled device (CCD).

24. The image detection system of claim 7, wherein the light representative of the biological sample comprises light provided by at least one of chemi-luminescence, fluorescence, chemi-fluorescence, photon excitation, phosphorescence, adsorption, and a quenching thereof.

25. The image detection apparatus of claim 7, wherein the apparatus has a depth of field greater than about 30 microns.

26. The image detection apparatus of claim 25, wherein the apparatus has a depth of field greater than about 50 microns

27. A method for detecting an image relating to a biological material, the method comprising:

providing a biological sample in a sampling position;

providing source light impinging on the biological sample such that, in response to such impinging light, light representative of the biological sample is produced along an emitted light path; and

positioning a light detector array in proximity to the biological sample and intersecting the emitted light path, and scanning the light detector array across a portion of the light representative of the biological sample.

28. The method of claim 27, wherein the source light is generated by a plurality of light emitting diodes or laser diodes.

29. The method of claim 28, further comprising providing source light at a first wavelength range and then providing source light at a second wavelength range that is substantially different from the first wavelength range.

30. The method of claim 27, wherein providing source light comprises providing excitation light that impinges on at least a portion of the biological material such that, in response to the excitation light, fluorescence representative of the biological sample is provided via the emitted light path for detection by the light detector.

31. The method of claim 30, further comprising filtering the source light to reduce interference between the source light and the light representative of the biological sample.

32. The method of claim 27, wherein the light detector array comprises a linear light detector array.

33. The method of claim 27, wherein the light detector array is scanned by translating the biological sample.

34. The method of claim 27, further comprising preventing light having the same or substantially the same frequency as the source light from impinging on the light detector.

35. A method for detecting an image relating to a biological sample, the method comprising:

providing a polynucleic acid chip having a sample of one or more polynucleic acid sequences thereon in a holder at a sampling position;

generating source light with one or more diodes;

impinging the source light on the sample to cause the sample to generate emitted light from at least a portion of the sample along an emitted light path, wherein the emitted light is generated by at least one of chemi-luminescence, fluorescence, chemi-fluorescence, phosphorescence, adsorption and photon excitation;

providing a light detector array in the emitted light path; and

translating the light detector array through at least a portion of the emitted light and detecting a plurality of images representative of the sample.

36. The method of claim 35, further comprising providing source light at a first wavelength range and providing source light at a second wavelength range that is substantially different from the first wavelength range.

37. The method of claim 35, further comprising filtering the source light to reduce interference between the source light and the emitted light generated from the biological sample.

38. The method of claim 35, further comprising preventing light having the same or substantially the same frequency as the source light from impinging on the light detector.

39. The method of claim 35, wherein the light path of the emitted light generated by the biological sample does not include any portion of the polynucleic acid chip.

40. A system for detecting a pattern of polynucleic acid hybridization in a biological sample, the system comprising:

a polynucleic acid chip holder to hold a polynucleic acid chip containing a sample in a sampling position;

a light source comprising one or more light emitting diodes positioned such that source light generated by the light source impinges on the sample and causes the portions of the sample to emit light along an emitted light path by at least one of chemi-luminescence, fluorescence, chemi-fluorescence, and photon excitation; and

a linear light detector array positionable in the emitted light path;

a scanning apparatus that translates the linear light detector across the light path to acquire a plurality of images representative of the sample.

41. The system of claim 40, wherein the light source comprises a first set of one or more diodes that generates light at a first wavelength range, and a second set of one or more diodes that generates light at a second wavelength range that is substantially different from the first wavelength range.

42. The system of claim 40, further comprising a source light filter adapted to reduce interference between the source light and the emitted light generated from the biological sample.

43. The system of claim 40, further comprising a lens assembly in the emitted light path such that at least a portion of the emitted light from the biological sample is focused on the light detector.

44. The system of claim 40, further comprising a chip transport mechanism having a plurality of chip holders and adapted to position a chip in a sampling position.

45. The system of claim 44, wherein the chip transport mechanism is a rotary mechanism.