Systems and methods for high-speed image scanning

• US 20080228404A1
• Filed: 08/24/2007
• Published: 09/18/2008
• Est. Priority Date: 08/25/2006
• Status: Active Grant

• 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.
• Granted Patent: US 7,826,977 B2
• Time in Patent Office: Days
• 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:

(a) adjusting an object using a positioning element;

(b) 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;

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

(d) detecting the portion of the image of the object using the sensor positioned relative to the first mirror and the second mirror.

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 sensor is a CMOS sensor.

4. The method of claim 1, wherein the sensor is a charge coupled device.

5. The method of claim 1, wherein the first mirror and the second mirror are located between an objective lens and a tube lens.

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

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

8. The method of claim 1, further comprising dividing the object into a plurality of fields of view.

9. The method of claim 8, further comprising dividing each of the plurality of fields of view into a plurality of high magnification views.

10. The method of claim 1, wherein the object is glued onto a substrate.

11. The method of claim 1, wherein the object is laminated onto a substrate.

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

13. The method of claim 12, further comprising:

adding microspheres having a predetermined diameter to the biological sample;

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

imaging the microspheres; and

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

14. The method of claim 13, wherein the microspheres have a diameter greater than about 0.6 micrometers.

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

16. The method of claim 13, wherein the microspheres have a diameter greater than the diameter of a filter substrate located adjacent to the biological sample.

17. 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.

18. The method of claim 17, wherein the color of the microspheres have an excitation wavelength of about 625 nm and an emission spectrum of about 645 nm.

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

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

21. The method of claim 13, wherein the microspheres are detected using a CMOS sensor.

22. The method of claim 13, wherein the microspheres are detected using a charge coupled device.

23. The method of claim 1, further comprising positioning at least a portion of an imaging element using a piezo nano-positioner.

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

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

26. 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.

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

28. The method of claim 1, 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.

29. The method of claim 28, 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.

30. The method of claim 28, wherein the image of the object is directed toward at least one of the first sensor and the second sensor using a dichroic mirror.

31. The method of claim 29, 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.

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

33. The method of claim 32, further comprising analyzing the plurality of images.

34. The method of claim 33, wherein at least one of the plurality of images is analyzed using cluster analysis.

35. The method of claim 34, 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.

36. The method of claim 28, wherein the second sensor is a 3 CMOS color sensor array.

37. The method of claim 28, further comprising illuminating the object using a light source.

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

39. The method of claim 37, wherein the light source is an LED.

40. The method of claim 37, wherein the light source is a lamp.

41. The method of claim 28, 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.

42. The method of claim 41, wherein the first light source is an LED and the second light source is an argon ion laser.

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

(a) an imaging element;

(b) 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;

(c) a first mirror positioned relative to the object;

(d) 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

(e) 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.

44. The apparatus of claim 43, wherein the first mirror and the second mirror are galvanometric mirrors.

45. The apparatus of claim 43, wherein the sensor is a CMOS sensor.

46. The apparatus of claim 43, wherein the sensor is a charge coupled device.

47. The apparatus of claim 43, further comprising an objective lens and a tube lens and wherein the first mirror and the second mirror are located between an objective lens and a tube lens.

48. The apparatus of claim 43, further comprising an imaging element.

49. The apparatus of claim 48, wherein a portion of the imaging element is a magnifying objective lens.

50. The apparatus of claim 43, wherein the positioning element is a linear actuator.

51. The apparatus of claim 43, wherein the positioning element is a stepper motor.

54. The apparatus of claim 43, wherein the object is a sample in a well in an array.

55. The apparatus of claim 54, 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 the wavelength emitted by the sample; and

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

56. The apparatus of claim 54, wherein the microspheres have a diameter greater than about 0.6 micrometers.

57. The apparatus of claim 54, wherein the microspheres have a diameter of about 10 micrometers.

58. The apparatus of claim 54, wherein the microspheres have a diameter greater than the diameter of a filter substrate included in the well.

59. The apparatus of claim 54, wherein the microspheres have a diameter similar to the diameter of the object to be imaged.

60. The apparatus of claim 54, wherein the microspheres have a fluorescence and wherein the fluorescence is different than the fluorescent used to image the sample.

61. The apparatus of claim 60, wherein the microspheres have an excitation wavelength of about 625 nm and an emission spectrum of about 645 nm.

62. The apparatus of claim 54, wherein the microspheres are illuminated using a lamp.

63. The apparatus of claim 54, wherein the microspheres are illuminated using a laser.

64. The apparatus of claim 54, wherein the microspheres are illuminated using a diode.

65. The apparatus of claim 54, wherein the microspheres are detected using a CMOS sensor.

66. The apparatus of claim 54, wherein the microspheres are detected using a charge coupled device.

67. The apparatus of claim 54, wherein the position of at least a portion of the imaging element is adjusted using a piezo nano-positioner.

68. The apparatus of claim 54, wherein the imaging element is a magnifying objective lens.

69. The apparatus of claim 54, further comprising a dichroic mirror, wherein the dichroic mirror is positioned in the path of the image of the sample to allow focusing on the microspheres.

70. The apparatus of claim 54, further comprising a dichroic mirror, wherein the dichroic mirror is positioned in the path of the image of the sample to allow imaging of the sample.

71. The apparatus of claim 43, wherein the sensor is a first sensor and further comprising:

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.

72. The apparatus of claim 71, wherein the first sensor is positioned along a first optical path and the second sensor is positioned along a second optical path.

73. The apparatus of claim 72, wherein the first optical path includes a first level of magnification and the second optical path includes a second level of magnification.

74. The apparatus of claim 71, wherein the at least one of the plurality of images is directed to the first sensor.

75. The apparatus of claim 74, further comprising a software program for analyzing the plurality of images.

76. The apparatus of claim 75, wherein the at least one of the plurality of images is analyzed using cluster analysis.

77. The apparatus of claim 76, 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.

78. The apparatus of claim 71, wherein the second sensor is a 3 CMOS color sensor array.

79. The apparatus of claim 71, further comprising a light source, wherein the light source illuminates the object.

80. The apparatus of claim 79, wherein the light source is an argon ion laser.

81. The apparatus of claim 79, wherein the light source is an LED.

82. The apparatus of claim 79, wherein the light source is a lamp.

83. The apparatus of claim 71, further comprising a first light source and a second light source.

84. The apparatus of claim 83, wherein the first light source is an LED and the second light source is an argon ion laser.

52. The apparatus of claim 52, wherein the first mirror and the second mirror are adjusted to provide images of the object from a plurality of fields of view.

53. The apparatus of claim 52, 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.

85. A system for high speed image scanning, comprising:

(a) an imaging means for providing a magnified image of an object;

(b) a positioning means controllable in an x direction and in a y direction, coupled to an object, wherein the positioning means adjusts the position of the object;

(c) a first reflecting means positioned relative to the object;

(d) a second reflecting means positioned relative to the object and the first reflecting means, wherein the first reflecting means directs at least one image in the x direction toward the second reflecting means and wherein the second reflecting means directs the at least one image of the object in the y direction toward a sensing means; and

(e) a processing means coupled to the positioning means, the first reflecting means and the second reflecting means, wherein the processing means controls the positioning means, the position of the first reflecting means and the position of the second reflecting means.