Light detection device

• US 6,071,748 A
• Filed: 04/17/1998
• Issued: 06/06/2000
• Est. Priority Date: 07/16/1997
• Status: Expired due to Term

• Current Assignee: Molecular Devices
• Original Assignee: LJL Biosystems, Inc. (Danaher Corporation)
• Inventors: Modlin, Douglas N., Edwards, Glenn R., Marquiss, Samuel A., Taylor, Michael T.
• Primary Examiner(s): Le, Long V.
• Application Number:
  US09/062,472
• Time in Patent Office: 781 Days
• Field of Search: 636/43, 636/68, 636/50, 636/164, 636/174, 636/805, 636/807, 636/809, 422/63, 422/65, 422/67, 422/82.05, 422/82.08, 422/52, 422/102, 422/104, 356/317, 356/318, 356/417, 250/458.1, 250/459.1, 250/461.1, 250/461.2, 250/428, 250/432 R, 250/491.1
• US Class Current: 436/174
• CPC Class Codes: B01J 2219/00315 Microtiter plates
• CPC Class Codes: B01J 2219/00317 Microwell devices, i.e. having large numbers of wells
• CPC Class Codes: B01J 2219/00605 the compounds being directly bound or immobilised to solid supports
• CPC Class Codes: B01J 2219/0061 The surface being organic
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00686 Automatic
• CPC Class Codes: B01J 2219/00691 using robots
• CPC Class Codes: B01J 2219/00707 separated from the reactor apparatus
• CPC Class Codes: B01L 2300/0829 Multi-well plates; Microtitration plates
• CPC Class Codes: B01L 9/523 for multisample carriers, e.g. used for microtitration plates
• CPC Class Codes: B82Y 15/00 Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors
• CPC Class Codes: C40B 60/14 for creating libraries
• CPC Class Codes: G01N 21/76 Chemiluminescence; Bioluminescence
• CPC Class Codes: Y10S 436/807 Apparatus included in process claim, e.g. physical support structures
• CPC Class Codes: Y10S 436/809 Multifield plates or multicontainer arrays
• CPC Class Codes: Y10T 436/11 Automated chemical analysis
• CPC Class Codes: Y10T 436/113332 with conveyance of sample along a test line in a container or rack
• CPC Class Codes: Y10T 436/114165 with step of insertion or removal from test line
• CPC Class Codes: Y10T 436/115831 Condition or time responsive
• CPC Class Codes: Y10T 436/25 : including sample preparation

Abstract

A high-throughput light detection instrument and method are described. Confocal optics structure enables exclusive light detection from a sensed volume in a composition.

Claims

1. An apparatus for detecting light transmitted from a composition, the apparatus comprising:

a stage for supporting a composition in an examination site, the composition being contained in a spatial volume lying between boundary interfaces located at different points along a Z-axis, wherein the Z-axis is substantially perpendicular to tie stage;

an automated registration device that automatically brings successive compositions and the examination site into register for successive analysis of the compositions;

a light source positioned to deliver light to the composition in the examination site;

a detector positioned to receive light transmitted from the composition in the examination site;

an optical relay structure located between the light source and the detector, the optical relay structure being capable of transmitting light substantially exclusively from a sensed volume of the composition, wherein the sensed volume is spaced substantially away from at least one of the boundary interfaces of the composition; and

a Z-axis optical adjustment device that automatically adjusts positions of the sensed volume along the Z-axis between and spaced away from the boundary interfaces.

2. The apparatus of claim 1, wherein the optical relay structure includes confocal optics elements substantially contained within an optics head positioned above or below the stage, the Z-axis optical adjustment device including a drive mechanism that moves the optics head relative to the Z-axis.

3. The apparatus of claim 1, wherein X, Y-axes are defined perpendicular to each other, perpendicular to the Z-axis and parallel to the stage, further comprising an X, Y-axes adjustment device that automatically adjusts the relative positions of the sensed volume and the composition along the X and Y axes.

4. The apparatus of claim 3, wherein the X, Y-axes adjustment device can be programmed to convey successive compositions to the examination site.

5. The apparatus of claim 4, wherein the successive compositions are contained in wells of a microplate.

6. The apparatus of claim 1, wherein the sensed volume is spaced substantially away from all the boundary interfaces.

7. The apparatus of claim 1, wherein the sensed volume has a waist region in a sample plane, and a Z-pick-up, the diameter of the waist region being approximately half the dimension of the Z-pick-up.

8. The apparatus of claim 1, wherein the optical relay structure includes an aperture substantially centered about the Z-axis and contained in an image plane conjugate to a sample plane intersecting the sensed volume.

9. The apparatus of claim 1, wherein the apparatus is capable of detecting light in at least two of the following types of assays:

luminescence intensity, photoluminescence polarization, chemiluminescence, photoluminescence lifetime absorbance, luminescence resonance energy transfer, and luminescence imaging.

10. The apparatus of claim 1, wherein the detector detects light of longer wavelength than the light delivered to the composition from the light source.

11. The apparatus of claim 1, further comprising an automated registration device controller for preprogramming the relative movement into registration of successive compositions and the examination site.

12. The apparatus of claim 1, wherein the light source is a lamp, or a light-emitting diode, or a laser, or a flash lamp, or a particle accelerator.

13. The apparatus of claim 1, wherein the light source is one of a plurality of light sources positioned at a source station, and further comprising a switching mechanism for interchangeably connecting different light sources to the optical relay structure for different applications.

14. The apparatus of claim 13, wherein the switching mechanism includes a shuttle having one side connected to a fiber optic element leading toward the stage, and a second side substantially facing the light sources, the shuttle being movable relative to the light sources, so that different light sources can be selectively transmitted through the fiber optic element to a composition at the examination site.

15. The apparatus of claim 13, further comprising a filter wheel connected to the source station, the filter wheel containing a plurality of filters and being rotatable about a rotational axis so that a given filter can be interchangeably interposed between different light sources and the optical relay structure.

16. The apparatus of claim 1, wherein the detector is a photomultiplier tube, or a photodiode, or a charge-coupled device.

17. The apparatus of claim 1, wherein successive compositions are held in adjacent wells in a microplate.

18. The apparatus of claim 17, wherein the number of wells in the microplate is 96, or 384, or 1536.

19. The apparatus of claim 1, wherein the optical relay structure simultaneously transmits light to the detector from compositions contained in a plurality of microplate wells.

20. The apparatus of claim 1, wherein the optical relay structure delivers light from the light source at an angle relative to the Z-axis exceeding the critical angle for establishing an evanescent field in the composition.

21. The apparatus of claim 1, wherein the optical relay structure delivers light from the light source at an angle relative to the Z-axis below the critical angle for establishing an evanescent field in the composition.

22. The apparatus of claim 1, wherein the sensed volume is diffraction limited.

23. The apparatus of claim 1, wherein the light source produces high-intensity, high-color temperature light.

24. The apparatus of claim 23, wherein the light source is a xenon arc lamp.

25. The apparatus of claim 1, wherein the automated registration device includes a transporter having a mechanism that releasably grips and moves a sample container to and from the examination site.

26. The apparatus of claim 25, wherein the transporter is dimensioned to grip and carry a microplate.

27. The apparatus of claim 25, wherein the transporter is the stage.

28. The apparatus of claim 27, further comprising

a housing, the transporter being movable along a path running between a loading position and the examination site, the loading position being substantially outside the housing, so that compositions can be easily loaded on and unloaded from the transporter when the transporter is in the loading position, and the transporter can deliver compositions to and from the examination site inside the housing before and after analysis.

29. The apparatus of claim 28, further comprising

a stacking unit mounted next to the loading position outside the housing, the stacking unit being configured to hold a plurality of sample containers, and to feed sample containers to the transporter one at a time.

30. The apparatus of claim 29, wherein the sample containers are croplates.

31. The apparatus of claim 27, wherein the transporter is open in the enter so that when the transporter is in the examination site, light can be transmitted rom below the stage to a composition in the examination site.

32. The apparatus of claim 1, wherein the optical relay structure includes a first aperture and a first lens positioned along a light path between the light source and the examination site or between the detector and the examination site.

33. The apparatus of claim 32, wherein the optical relay structure includes a second aperture and a second lens, the first aperture and the first lens being positioned along a light path between the light source and the examination site, the second aperture and the second lens being positioned along a light path between the detector and the examination site so that light is transmitted substantially exclusively to and from the same sensed volume in a composition at the examination site.

34. The apparatus of claim 32, wherein the optical relay structure cludes at least one fiber optic element, the first aperture being defined by the dimension of an end of the fiber optic element.

35. The apparatus of claim 32, wherein the first aperture is defined in a ask structure at an end of a fiber optic element.

36. The apparatus of claim 32, wherein the optical relay structure includes a second aperture, the first and second apeitures having different dimensions, and an aperture switching mechanism for interchangeably positioning a selected aperture in the light path leading to or from a sensed volume in a composition at the examination site, so that the size of the sensed volume can be altered by switching between the first and second apertures.

37. The apparatus of claim 32, wherein the diameter of the first aperture is adjustable.

38. The apparatus of claim 32, wherein the first aperture and the first lens are contained in a first optics head positioned near the stage.

39. The apparatus of claim 38, wherein the first optics head is positioned above the stage.

40. The apparatus of claim 38, wherein the first optics head is positioned below the stage.

41. The apparatus of claim 38, wherein the first optics head has a light entrance port optically connected to the light source, and a light exit port optically connected to the detector, so that illumination and detection of light transmitted from a composition in the examination site can be carried out fiom the same side of the stage.

42. The apparatus of claim 41, wherein the light entrance port and the light exist port are configured to transmit light in directions that are substantially perpendicular to each other.

43. The apparatus of claim 42, wherein the first optics head includes a planar beam splitter oriented at 45 degree angles to the directions of light transmission.

44. The apparatus of claim 39, wherein the optical relay structure includes a second optics head positioned below the stage, each optics head having a light entrance port optically connected to the light source, and a light exit port optically connected to the detector, and a switch control mechanism capable of interchangeably configuring any one of the following light transmission routes to and from a sensed volume in a composition located at the examination site:

(a) top-illumination and top-detection, (b) top-illumination and bottom-detection, (c) bottom-illumination and top-detection, and (d) bottom-illumination and bottom-detection.

77. The apparatus of claim 44, wherein each optics head is independently adjustable along the Z-axis.

45. An apparatus for detecting light transmitted from a composition, the apparatus comprising:

a stage for supporting a composition in a spatial volume having boundary interfaces located along surfaces in a microplate well;

a light source positioned to deliver light from the light source into the composition;

a detector;

an optical relay structure that is capable of transmitting light substantially exclusively from a sensed volume of the composition to the detector, wherein the sensed volume is spaced substantially away from the boundary interfaces of the composition; and

a Z-axis optical adjustment device, wherein a Z-axis is defined perpendicular to the stage, the Z-axis optical adjustment device that automatically adjusts positions the sensed volume along the Z-axis in the composition between and spaced away from the boundary interfaces.

46. The apparatus of claim 45, wherein X, Y-axes are defined perpendicular to each other, perpendicular to the Z-axis and parallel to the stage, further comprising an X, Y-axes adjustment device that automatically adjusts the relative positions of the sensed volume and the composition along the X and Y-axes.

47. The apparatus of claim 45, wherein a Z-axis is defined substantially perpendicular to the stage of an examination site, the optical relay structure including an aperture substantially centered about the Z-axis and contained in an image plane conjugate to a sample plane intersecting the sensed volume.

48. The apparatus of claim 45, wherein the apparatus is capable of detecting light in at least two of the following types of assays:

luminescence intensity, photoluminescence polarization, cheiniluminescence, photoluminescence lifetime absorbance, luminescence resonance energy transfer, luminescence imaging.

49. The apparatus of claim 45, wherein the detector detects light of longer wavelength than the light delivered to the composition from the light source.

50. The apparatus of claim 45, wherein the light source is a lamp, or a light-emitting diode, or a laser, or a flash lamp, or a particle accelerator.

51. The apparatus of claim 45, wherein the detector is a photomultiplier tube, or a photodiode, or a charge-coupled device.

52. The apparatus of claim 45, wherein the microplate well is contained in a microplate, wherein the number of wells in the microplate is 96, or 384, or 1536.

53. The apparatus of claim 45, wherein the microplate well containing the composition is located at an examination site, the optical relay structure including a first aperture and a first lens positioned along a light path between the detector and the examination site.

54. The apparatus of claim 45, wherein the optical relay structure includes a second aperture and a second lens positioned along a light path running between the light source and the examination site.

55. The apparatus of claim 53, wherein the optical relay structure includes at least one fiber optic element, the first aperture being defined by the dimension of an end of the fiber optic element.

56. The apparatus of claim 53, wherein the first aperture is defined in a mask structure at an end of a fiber optic element.

57. The apparatus of claim 53, wherein the optical relay structure includes a second aperture, the first and second apertures having different dimensions, and an aperture switching mechanism for interchangeably positioning a selected aperture in the light path leading to or from a sensed volume in a composition at the examination site, so that the size of the sensed volume can be altered by switching between the first and second apertures.

58. The apparatus of claim 53, wherein the first aperture and the first lens are contained in a first optics head positioned near the stage.

59. The apparatus of claim 58, wherein the first optics head is positioned above the stage.

60. The apparatus of claim 58, wherein the first optics head is positioned below the stage.

61. The apparatus of claim 58, wherein the first optics head has a light entrance port optically connected to the light source, and a light exit port optically connected to the detector, so that illumination and detection of light transmitted from a composition in the examination site can be carried out fiom the same side of the stage.

62. The apparatus of claim 61, wherein the light entrance port and the light exit port are configured to transmit light in directions that are substantially perpendicular to each other.

63. The apparatus of claim 62, wherein the first optics head includes a planar beamsplitter oriented at 45-degree angles to the directions of light transmission.

64. The apparatus of claim 59, wherein the optical relay structure includes a second optics head positioned below the stage, each optics head having a light entrance port optically connected to the light source, and a light exit port optically connected to the detector, and a switch control mechanism capable of interchangeably configuring any one of the following light transmission routes to and from a sensed volume in a composition located at the examination site:

(a) top-illumination and top-detection, (b) top-illumination and bottom-detection, (c) bottom-illumination and top-detection, and (d) bottom-illumination and bottom-detection.

65. An apparatus for detecting light transmitted from a composition, the apparatus comprising:

a stage for suppoing a composition in a spatial volume having boundary interfaces located along surfaces in a microplate well;

a light source;

an optical relay structure that is capable of delivering light from the light source substantially exclusively to a sensed volume of the composition, wherein the sensed volume is spaced substantially away from the boundary interfaces of the composition;

a detector positioned to receive light transmitted from the sensed volume of the composition; and

a Z-axis optical adjustment device, wherein a Z-axis is defined perpendicular to the stage, the Z-axis optical adjustment device that automatically adjusts positions of the sensed volume along the Z-axis between and spaced away from the boundary interfaces of the composition.

66. An apparatus for detecting photoluminescence emitted by a sample, the apparatus comprising:

a stage configured to hold a microplate having an array of sample wells;

a light source located at one end of a first light path leading toward the stage and configured to deliver light to induce photoluminescence in a sample contained within boundary interfaces in at least one of the sample wells along a Z-axis perpendicular to the stage;

a detector located at one end of a second light path leading toward the stage and configured to detect photolumiescence emitted by the sample;

a confocal optics element operatively positioned along the first light path or the second light pat, and configured substantially to prevent light from focusing outside a pre-selected sensed volume inside the sample, or configured substantially to prevent detection of photoluminescence originating outside the sensed volume; and

an optical adjustment device that automatically adjusts positions of the sensed volume along the Z-axis inside the sample between and spaced away from the boundary interfaces.

67. An apparatus for detecting luminescence emitted by a sample, the apparatus comprising:

a stage configured to hold a sample inside boundary interfaces along a Z-axis perpendicular to the stage at an examination site for analysis;

an alignment device configured to adjust the relative positions of the stage and the examination site, so that the sample and the examination site can be brought into alignment before detection of luninescence, and removed from alignment after such detection;

an alignment device controller configured automatically to control the alignment device, so that successive samples can be thought into alignment with the examination site for detection of luminescence;

an optical focusing structure configured sequentially to focus luminescence from the portion of the sample positioned in a sample plane into a series of image planes, wherein the image planes include at least one intermediate image plane and one terminal image plane;

a detector configured to detect luminescence emitted by the sample, wherein the detector is positioned substantially in the terminal image plane;

an emission confocal optics element configured substantially to prevent detection of luminescence originating outside a sensed volume intersecting the sample plane, wherein the emission confocal optics element is positioned between the sample and the detector substantially in an intermediate image plane; and

a sensed volume positioning mechanism that automatically adjusts positions of the sensed volume along the Z-axis inside the sample between and spaced away from the boundary interfaces.

68. The apparatus of claim 67, wherein the optical focusing structure includes at least one lens positioned along a light path leading from the sample plane to the detector.

69. The apparatus of claim 67, wherein the emission confocal optics element defines an aperture.

70. The apparatus of claim 69, wherein the aperture is adjustable.

71. The apparatus of claim 67, further comprising:

a light source configured to deliver light to induce luminescence in the sample; and

an excitation confocal optics element configured substantially to prevent light from the light source from focusing outside the portion of the sample plane contained in the sensed volume;

wherein the excitation confocal optics element is positioned substantially in an intermediate image plane located between the light source and the sample; and

wherein there are at least two intermediate image planes, with the emission confocal optics element and the excitation confocal optics element positioned in different image planes.

72. A method of detecting light transmitted from a composition, the method comprising:

automatically bringing into register a succession of compositions and an examination site for successive analysis of the compositions, wherein each composition is contained between top, bottom, and side boundary interfaces located at different points along a Z-axis;

transmitting light into a composition at the examination site;

autmatically adjusting the position of a sensed volume along the Z-axis between and spaced away from the boundary interfaces; and

detecting light substantially exclusively from the sensed volume in the composition.

75. The method of claim 72 further comprising the step of

spacing the sensed volume away from at least one of the boundary interfaces.

76. The method of claim 72 further comprising the step of

spacing the sensed volume away from both of the top and bottom boundary interfaces.

73. A high-throughput method of detecting light transmitted from a succession of samples, the method comprising:

automatically and sequentially aligning a succession of samples with a Z-axis at an examination site, a sample being contained within boundary interfaces;

for each sample, focusing light originating from the waist region of a sensed volume into a series of image planes, beginning with at least one intermediate image plane and terminating with a terminal image plane;

positioning a detector in the terminal image plane to detect light transmitted by the sample;

positionng an emission confocal optics element in an intermediate image plane, wherein the emission confocal optics element is configured substantially to prevent detection of light originating outside the sensed volume; and

automatically adjusting the position of the sensed volume along the Z-axis inside the sample between and spaced away from the boundary interfaces.

74. The high-throughout method of claim 73, further comprising:

positioning a light source to induce emission of light by the sample; and

positioning an excitation confocal optics element in an intermediate image plane, wherein the excitation confocal optics element is configured substantially to prevent light from the light source from focusing outside the waist region of the sensed volume.