Systems and methods for high-speed image scanning

• US 7,826,977 B2
• Filed: 08/24/2007
• Issued: 11/02/2010
• Est. Priority Date: 08/25/2006
• Status: Expired due to Fees

• Current Assignee: Trustees Of Columbia University In The City Of New York (Columbia University)
• Original Assignee: Trustees Of Columbia University In The City Of New York (Columbia University)
• Inventors: Brenner, David J., Randers-Pehrson, Gerhard, Garty, Guy, Lyulko, Oleksandra V.
• Primary Examiner(s): Wachsman; Hal D
• Assistant Examiner(s): O MALLEY, MARY CATHERINE
• Application Number:
  US11/895,470
• Publication Number:
  US 20080228404A1
• Time in Patent Office: 1,166 Days
• Field of Search: 702/19, 702 21- 22, 702 27- 28, 702/31, 356317-318, 356/417, 250/458.1, 250/459.1, 250/461.1, 250/461.2, 250/491.1, 250/504.R
• US Class Current: 702/19
• CPC Class Codes: C40B 30/10 by measuring physical properties, e.g. mass
• CPC Class Codes: C40B 60/12 for screening libraries

Abstract

Systems and methods for high-speed image scanning are disclosed herein One aspect of the invention is directed to a method for high speed image scanning. The method for high speed image scanning includes adjusting an object using a positioning element; directing a portion of an image of the object toward a sensor by positioning a first mirror relative to the object, and by positioning a second mirror relative to the object and the first mirror; controlling the positioning element, the position of the first mirror and the position of the second mirror using a processor; and detecting the portion of the image of the object using the sensor positioned relative to the first mirror and the second mirror. In accord with this method, the first mirror directs the portion of the image of the object in a first direction and the second mirror directs the portion of the image of the object in a second direction.

Claims

1. A method for high speed image scanning, comprising:

adjusting an object using a positioning element;

directing a portion of an image of the object toward a sensor by positioning a first mirror relative to the object, wherein the first mirror directs the portion of the image of the object in a first direction, and by positioning a second mirror relative to the object and the first mirror, wherein the second mirror directs the portion of the image of the object in a second direction;

controlling the positioning element, the position of the first mirror and the position of the second mirror using a processor; and

detecting the portion of the image of the object using the sensor positioned relative to the first mirror and the second mirror, wherein the sensor is at least one of a CMOS sensor and a charge coupled device.

2. The method of claim 1, wherein the first mirror and the second mirror are galvanometric mirrors.

3. The method of claim 1, wherein the positioning element is a linear actuator.

4. The method of claim 1, wherein the positioning element is a stepper motor.

5. The method of claim 1, wherein the object is a biological sample.

6. The method of claim 5, further comprising:

adding microspheres having a predetermined diameter to the biological sample;

illuminating the microspheres such that they emit a wavelength different than a wavelength emitted by the biological sample;

imaging the microspheres; and

adjusting the position of the imaging element to bring the biological sample into focus.

7. The method of claim 6, wherein the microspheres are detected using a CMOS sensor.

8. The method of claim 6, wherein the microspheres are detected using a charge coupled device.

9. A method for high speed image scanning, comprising:

adjusting an object using a positioning element;

directing a portion of an image of the object toward a sensor by positioning a first mirror relative to the object, wherein the first mirror directs the portion of the image of the object in a first direction, and by positioning a second mirror relative to the object and the first mirror, wherein the second mirror directs the portion of the image of the object in a second direction;

controlling the positioning element, the position of the first mirror and the position of the second mirror using a processor; and

detecting the portion of the image of the object using the sensor positioned relative to the first mirror and the second mirror, wherein the first mirror and the second mirror are located between an objective lens and a tube lens.

10. A method for high speed image scanning, comprising:

adjusting an object using a positioning element;

directing a portion of an image of the object toward a sensor by positioning a first mirror relative to the object, wherein the first mirror directs the portion of the image of the object in a first direction, and by positioning a second mirror relative to the object and the first mirror, wherein the second mirror directs the portion of the image of the object in a second direction;

controlling the positioning element, the position of the first mirror and the position of the second mirror using a processor;

detecting the portion of the image of the object using the sensor positioned relative to the first mirror and the second mirror; and

dividing the object into a plurality of fields of view.

11. The method of claim 10, further comprising dividing each of the plurality of fields of view into a plurality of high magnification views.

12. A method for high speed image scanning, comprising:

adjusting an object using a positioning element;

directing a portion of an image of the object toward a sensor by positioning a first mirror relative to the object, wherein the first mirror directs the portion of the image of the object in a first direction, and by positioning a second mirror relative to the object and the first mirror, wherein the second mirror directs the portion of the image of the object in a second direction;

controlling the positioning element, the position of the first mirror and the position of the second mirror using a processor; and

detecting the portion of the image of the object using the sensor positioned relative to the first mirror and the second mirror, wherein the object is at least one of glued onto a substrate and laminated onto a substrate.

13. A method for high speed image scanning, comprising:

adjusting an object using a positioning element, wherein the object is a biological sample;

directing a portion of an image of the object toward a sensor by positioning a first mirror relative to the object, wherein the first mirror directs the portion of the image of the object in a first direction, and by positioning a second mirror relative to the object and the first mirror, wherein the second mirror directs the portion of the image of the object in a second direction;

controlling the positioning element, the position of the first mirror and the position of the second mirror using a processor;

detecting the portion of the image of the object using the sensor positioned relative to the first mirror and the second mirror;

adding microspheres having a predetermined diameter to the biological sample;

illuminating the microspheres such that they emit a wavelength different than a wavelength emitted by the biological sample;

imaging the microspheres; and

adjusting the position of an imaging element to bring the biological sample into focus, wherein the microspheres have a diameter greater than at least one of about 0.6 micrometers and the diameter of a filter substrate located adjacent to the biological sample.

14. The method of claim 13, wherein the microspheres have a diameter of about 10 micrometers.

15. The method of claim 13, wherein the microspheres are fluorescent and wherein the fluorescence of the microspheres is different than the fluorescence used to image the sample.

16. The method of claim 15, wherein the color of the microspheres have an excitation wavelength of about 625 nm and an emission spectrum of about 645 nm.

17. The method of claim 13, wherein the microspheres are illuminated using a lamp.

18. The method of claim 13, wherein the microspheres are illuminated using a laser.

19. The method of claim 13, further comprising positioning a dichroic mirror in the path of the image of the sample to allow focusing on the microspheres or imaging of the sample.

20. The method of claim 13, wherein the microspheres are fluorescent.

21. A method for high speed image scanning, comprising:

adjusting an object using a positioning element;

directing a portion of an image of the object toward a sensor by positioning a first mirror relative to the object, wherein the first mirror directs the portion of the image of the object in a first direction, and by positioning a second mirror relative to the object and the first mirror, wherein the second mirror directs the portion of the image of the object in a second direction;

controlling the positioning element, the position of the first mirror and the position of the second mirror using a processor;

detecting the portion of the image of the object using the sensor positioned relative to the first mirror and the second mirror; and

positioning at least a portion of an imaging element using a piezo nano-positioner.

22. The method of claim 21, wherein the imaging element is a magnifying objective lens.

23. The method of claim 21, wherein the imaging element is a magnifying objective lens and a tube lens.

24. A method for high speed image scanning, comprising:

adjusting an object using a positioning element;

directing a portion of an image of the object toward a sensor by positioning a first mirror relative to the object, wherein the first mirror directs the portion of the image of the object in a first direction, and by positioning a second mirror relative to the object and the first mirror, wherein the second mirror directs the portion of the image of the object in a second direction;

controlling the positioning element, the position of the first mirror and the position of the second mirror using a processor; and

detecting the portion of the image of the object using the sensor positioned relative to the first mirror and the second mirror, wherein the sensor is a first sensor and further comprising directing the image of the object toward at least one of the first sensor and a second sensor.

25. The method of claim 24, wherein the image of the object is directed along a first optical path to the first sensor and along a second optical path to the second sensor.

26. The method of claim 25, further comprising magnifying a first image along the first optical path to a first level of magnification and magnifying a second image along the second optical path to a second level of magnification.

27. The method of claim 24, wherein the image of the object is directed toward at least one of the first sensor and the second sensor using a dichroic mirror.

28. The method of claim 24, further comprising directing a plurality of images of the object to the first sensor.

29. The method of claim 28, further comprising analyzing the plurality of images.

30. The method of claim 29, wherein at least one of the plurality of images is analyzed using cluster analysis.

31. The method of claim 30, further comprising:

analyzing the results of the cluster analysis;

comparing the results of the cluster analysis to a threshold; and

directing an image of the object to the second sensor when the results of the cluster analysis exceed the threshold.

32. The method of claim 24, wherein the second sensor is a 3 CMOS color sensor array.

33. The method of claim 24, further comprising illuminating the object using a light source.

34. The method of claim 33, wherein the light source is an argon ion laser.

35. The method of claim 33, wherein the light source is an LED.

36. The method of claim 33, wherein the light source is a lamp.

37. The method of claim 24, further comprising illuminating the object for a first instance using a first light source and illuminating the object for a second instance using a second light source.

38. The method of claim 37, wherein the first light source is an LED and the second light source is an argon ion laser.

39. An apparatus for high speed image scanning, comprising:

an imaging element;

a positioning element controllable in an x direction and in a y direction, coupled to an object, wherein the positioning element adjusts the position of the object;

a first mirror positioned relative to the object;

a second mirror positioned relative to the object and the first mirror, wherein the first mirror directs at least one image of the object in the x direction toward the second mirror and wherein the second mirror directs the at least one image of the object in the y direction toward a sensor; and

a processor coupled to the positioning element, the first mirror and the second mirror, wherein the processor controls the positioning element, the position of the first mirror and the position of the second mirror, wherein the sensor is at least one of a CMOS sensor and a charge coupled device.

40. The apparatus of claim 39, wherein the first mirror and the second mirror are galvanometric mirrors.

41. The apparatus of claim 39, wherein the positioning element is a linear actuator.

42. The apparatus of claim 39, wherein the positioning element is a stepper motor.

43. The apparatus of claim 39, wherein the first mirror and the second mirror are adjusted to provide images of the object from a plurality of fields of view.

44. The apparatus of claim 43, wherein the first mirror and the second mirror are adjusted to provide a plurality of high magnification images of the object from each of the plurality of fields of view.

45. The apparatus of claim 39, wherein the object is a sample in a well in an array.

46. The apparatus of claim 45, further comprising:

a plurality of microspheres having a predetermined diameter;

wherein the microspheres are added to the sample;

a light source, wherein the light source illuminates the microspheres such that the microspheres emit a wavelength different than a wavelength emitted by the sample; and

wherein the sensor detects images of the microspheres and the positioning element adjusts the position of the imaging element to bring the sample into focus.

47. The apparatus of claim 45, wherein the imaging element is a magnifying objective lens.

48. The apparatus of claim 45, further comprising a dichroic mirror, wherein the dichroic mirror is positioned in a path of the image of the sample to allow imaging of the sample.

49. An apparatus for high speed image scanning, comprising:

an imaging element;

a positioning element controllable in an x direction and in a y direction, coupled to an object, wherein the positioning element adjusts the position of the object;

a first mirror positioned relative to the object;

a second mirror positioned relative to the object and the first mirror, wherein the first mirror directs at least one image of the object in the x direction toward the second mirror and wherein the second mirror directs the at least one image of the object in the y direction toward a sensor;

a processor coupled to the positioning element, the first mirror and the second mirror, wherein the processor controls the positioning element, the position of the first mirror and the position of the second mirror; and

an objective lens and a tube lens, wherein the first mirror and the second mirror are located between the objective lens and the tube lens.

50. An apparatus for high speed image scanning, comprising:

an imaging element;

a positioning element controllable in an x direction and in a y direction, coupled to an object, wherein the positioning element adjusts the position of the object;

a first mirror positioned relative to the object;

a second mirror positioned relative to the object and the first mirror, wherein the first mirror directs at least one image of the object in the x direction toward the second mirror and wherein the second mirror directs the at least one image of the object in the y direction toward a sensor; and

a processor coupled to the positioning element, the first mirror and the second mirror, wherein the processor controls the positioning element, the position of the first mirror and the position of the second mirror, wherein a portion of the imaging element is a magnifying objective lens.

51. An apparatus for high speed image scanning, comprising:

an imaging element;

a positioning element controllable in an x direction and in a y direction, coupled to an object, wherein the positioning element adjusts the position of the object, wherein the object is a sample in a well in an array, and wherein the positioning element adjusts the position of the imaging element to bring the sample into focus;

a first mirror positioned relative to the object;

a second mirror positioned relative to the object and the first mirror, wherein the first mirror directs at least one image of the object in the x direction toward the second mirror and wherein the second mirror directs the at least one image of the object in the y direction toward a sensor;

a processor coupled to the positioning element, the first mirror and the second mirror, wherein the processor controls the positioning element, the position of the first mirror and the position of the second mirror;

a plurality of microspheres having a predetermined diameter, wherein the microspheres are added to the sample; and

a light source configured to illuminate the microspheres such that the microspheres emit a wavelength different than a wavelength emitted by the sample,

wherein the sensor detects images of the microspheres and wherein the microspheres have a diameter greater than at least one of about 0.6 micrometers and the diameter of a filter substrate included in the well.

52. The apparatus of claim 51, wherein the microspheres have a diameter of about 10 micrometers.

53. The apparatus of claim 51, wherein the microspheres further have a fluorescence and wherein the fluorescence is different than the fluorescent used to image the sample.

54. The apparatus of claim 53, wherein the microspheres further have an excitation wavelength of about 625 nm and an emission spectrum of about 645 nm.

55. The apparatus of claim 51, wherein the microspheres are illuminated using a lamp.

56. The apparatus of claim 51, wherein the microspheres are illuminated using a laser.

57. The apparatus of claim 51, wherein the microspheres are illuminated using a diode.

58. The apparatus of claim 51, further comprising a dichroic mirror positioned in a path of the image of the sample to allow focusing on the microspheres.

59. An apparatus for high speed image scanning, comprising:

an imaging element;

a positioning element controllable in an x direction and in a y direction, coupled to an object, wherein the positioning element adjusts the position of the object, wherein the object is a sample in a well in an array, and wherein the positioning element adjusts the position of the imaging element to bring the sample into focus;

a first mirror positioned relative to the object;

a second mirror positioned relative to the object and the first mirror, wherein the first mirror directs at least one image of the object in the x direction toward the second mirror and wherein the second mirror directs the at least one image of the object in the y direction toward a sensor;

a processor coupled to the positioning element, the first mirror and the second mirror, wherein the processor controls the positioning element, the position of the first mirror and the position of the second mirror;

a plurality of microspheres having a predetermined diameter, wherein the microspheres are added to the sample; and

a light source configured to illuminate the microspheres such that the microspheres emit a wavelength different than a wavelength emitted by the sample,

wherein the sensor detects images of the microspheres and wherein the microspheres have a diameter similar to the diameter of the object to be imaged.

60. An apparatus for high speed image scanning, comprising:

an imaging element;

a positioning element controllable in an x direction and in a y direction, coupled to an object, wherein the positioning element adjusts the position of the object, wherein the object is a sample in a well in an array, and wherein the positioning element adjusts the position of the imaging element to bring the sample into focus;

a first mirror positioned relative to the object;

a second mirror positioned relative to the object and the first mirror, wherein the first mirror directs at least one image of the object in the x direction toward the second mirror and wherein the second mirror directs the at least one image of the object in the y direction toward a sensor;

a processor coupled to the positioning element, the first mirror and the second mirror, wherein the processor controls the positioning element, the position of the first mirror and the position of the second mirror;

a plurality of microspheres having a predetermined diameter, wherein the microspheres are added to the sample; and

a light source configured to illuminate the microspheres such that the microspheres emit a wavelength different than a wavelength emitted by the sample,

wherein the sensor detects images of the microspheres and wherein the microspheres are detected using at least one of a CMOS sensor and a charge coupled device.

61. An apparatus for high speed image scanning, comprising:

an imaging element;

a positioning element controllable in an x direction and in a y direction, coupled to an object, wherein the positioning element adjusts the position of the object and wherein the object is a sample in a well in an array;

a first mirror positioned relative to the object;

a second mirror positioned relative to the object and the first mirror, wherein the first mirror directs at least one image of the object in the x direction toward the second mirror and wherein the second mirror directs the at least one image of the object in the y direction toward a sensor; and

a processor coupled to the positioning element, the first mirror and the second mirror, wherein the processor controls the positioning element, the position of the first mirror and the position of the second mirror, wherein the position of at least a portion of the imaging element is adjusted using a piezo nano-positioner.

62. An apparatus for high speed image scanning, comprising:

an imaging element;

a positioning element controllable in an x direction and in a y direction, coupled to an object, wherein the positioning element adjusts the position of the object;

a first mirror positioned relative to the object;

a second mirror positioned relative to the object and the first mirror, wherein the first mirror directs at least one image of the object in the x direction toward the second mirror and wherein the second mirror directs the at least one image of the object in the y direction toward a first sensor;

a second sensor, wherein the first mirror directs an image in the x direction toward the second mirror and wherein the second mirror directs the image in the y direction toward at least one of the first sensor and the second sensor; and

a processor coupled to the positioning element, the first mirror and the second mirror, wherein the processor controls the positioning element, the position of the first mirror and the position of the second mirror.

63. The apparatus of claim 62, wherein the first sensor is positioned along a first optical path and the second sensor is positioned along a second optical path.

64. The apparatus of claim 63, wherein the first optical path includes a first level of magnification and the second optical path includes a second level of magnification.

65. The apparatus of claim 62, wherein at least one of a plurality of images is directed to the first sensor.

66. The apparatus of claim 65, further comprising a software program for analyzing the plurality of images.

67. The apparatus of claim 66, wherein the at least one of the plurality of images is analyzed using cluster analysis.

68. The apparatus of claim 67, wherein the results of the cluster analysis are compared to a threshold, and wherein when the results of the cluster analysis exceed the threshold an image of the object is directed to the second sensor.

69. The apparatus of claim 62, wherein the second sensor is a 3 CMOS color sensor array.

70. The apparatus of claim 62, further comprising a light source, wherein the light source illuminates the object.

71. The apparatus of claim 70, wherein the light source is an argon ion laser.

72. The apparatus of claim 70, wherein the light source is an LED.

73. The apparatus of claim 70, wherein the light source is a lamp.

74. The apparatus of claim 62, further comprising a first light source and a second light source.

75. The apparatus of claim 74, wherein the first light source is an LED and the second light source is an argon ion laser.