Detection device for light transmitted from a sensed volume

• US 6,469,311 B1
• Filed: 07/31/2000
• Issued: 10/22/2002
• Est. Priority Date: 07/16/1997
• Status: Expired due to Term

• Current Assignee: Molecular Devices
• Original Assignee: Molecular Devices
• Inventors: Lemberg, Vladimir G., Modlin, Douglas N.
• Primary Examiner(s): LE, QUE TAN
• Application Number:
  US09/629,599
• Time in Patent Office: 813 Days
• Field of Search: 250/576, 250/573, 250/559.4, 250/461.2, 250/432.R, 250/338.5, 356/343, 356/317, 356/318, 356/417, 422/63, 422/82.08
• US Class Current: 250/576
• CPC Class Codes: G01N 2021/6419 Excitation at two or more wavelengths
• CPC Class Codes: G01N 2021/6421 Measuring at two or more wavelengths
• CPC Class Codes: G01N 21/6408 with measurement of decay time, time resolved fluorescence
• CPC Class Codes: G01N 21/6445 Measuring fluorescence polarisation
• CPC Class Codes: G01N 21/6452 Individual samples arranged in a regular 2D-array, e.g. multiwell plates
• CPC Class Codes: G01N 21/76 Chemiluminescence; Bioluminescence

Abstract

A high-throughput light detection instrument and method are described. In some embodiments, switch mechanisms and optical relay structures permit different light sources and/or detectors to be selected for different applications. In other embodiments, switch mechanisms and optical paths permit top/bottom illumination and/or top/bottom detection, or combinations thereof.

Claims

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

a stage for supporting a sample in a microplate well having a diameter X

a light source positioned to deliver light to the sample,

a detector positioned to receive light transmitted from the sample, and

a confocal optics device that positions a focal spot in the sample, the focal spot having a diameter of at least about 0.3X.

2. An apparatus for detecting light transmitted from a sample, the apparatus comprising

a stage for supporting a sample in a microplate well having a diameter X,

a light source positioned to deliver light to the sample,

a detector positioned to receive light transmitted from the sample, and

an optical relay structure that positions a sensed volume in the sample, the sensed volume being characterized by a waist region having a diameter D and a Z-pick-up having a height of at least about 2D.

3. An apparatus for detecting light transmitted from a sample, the apparatus comprising

a stage for supporting a sample in a microplate well, the sample having boundary interfaces defined by walls of the microplate well and an upper gas interface,

a light source positioned to deliver light to the sample,

a detector positioned to receive light transmitted from the sample,

a sample handling mechanism capable of automatically detecting light serially from microplate wells in a density of at least about 384 wells per plate, and

an optical relay structure that positions a sensed volume in the sample, the sensed volume being spaced away from the boundary interfaces.

4. The apparatus of claim 3, wherein the sample handling mechanism is capable of automatically detecting light serially from microplate wells in a density of at least about 1536 wells per plate.

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

a stage configured to support the composition in an examination site,

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 along an excitation optical axis, and

a detector positioned to receive light transmitted from the composition in the examination site along an emission optical axis,

wherein the excitation optical axis and the emission optical axis are not colinear.

6. The apparatus of claim 5, wherein the excitation optical axis is angled relative to the emission optical axis.

7. The apparatus of claim 6, wherein the excitation optical axis is angled to create total internal reflection in the composition.

8. A light detection system comprising

a stage for supporting a microplate having a plurality of wells, each well having a maximum volume capacity of about 55 microliters or less,

a light source,

a detector,

an optical relay structure that transmits light from the light source to a sample contained in a well in the microplate, and from the sample to the detector, and

a Z-axis adjustment device that alters location of a sensed volume inside the sample to optimize signal-to-background performance of the system.

9. The system of claim 8, wherein the microplate has at least about 384 wells.

10. The system of claim 8, wherein each well in the microplate has a maximum volume capacity of about 20 microliters or less.

11. The system of claim 8, wherein each well in the microplate has a substantially frustoconical shape.

12. The system of claim 8, wherein the optical relay structure includes confocal optics.

13. The system of claim 8, wherein the optical relay structure generates a sensed volume in the sample that substantially avoids boundary interfaces of the sample.

14. The system of claim 8, wherein the optical relay structure generates a sensed volume that extracts maximum signal from the sample.

15. The system of claim 8, wherein the optical relay structure generates a sensed volume, at least a portion of the sensed volume having a shape that is substantially frustoconical.

16. The system of claim 8, wherein the optical relay structure generates a sensed volume, at least a portion of the sensed volume having a shape that substantially corresponds to the shape of the well.

17. The system of claim 8, wherein the microplate has a frame portion that is compatible with robotics that are designed to handle standard microplates having 96 wells per plate.

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

means for supporting the composition in an examination site, the composition being contained in a spatial volume delineated by boundary interfaces,

means for automatically bringing successive compositions and the examination site into register for successive analysis of the compositions,

means for producing light and delivering the light to the composition in the examination site,

means for transmitting light substantially exclusively from a sensed volume of the composition,

means for automatically positioning the sensed volume substantially away from at least one of the boundary interfaces of the composition, and

means for detecting the light transmitted substantially exclusively from the sensed volume of the composition.

19. The apparatus of claim 18, wherein the successive compositions are held in adjacent wells in a microplate.

20. The apparatus of claim 18, wherein the means for producing light includes a light source selected from the group consisting of flash lamps, arc lamps, incandescent lamps, fluorescent lamps, electroluminescence devices, lasers, laser diodes, and light-emitting diodes (LEDs).

21. The apparatus of claim 18, wherein the means for transmitting light substantially exclusively from a sensed volume include a confocal optics element.

22. The apparatus of claim 18, wherein the means for positioning the sensed volume automatically positions the sensed volume between the boundary interfaces according to stored parameters relating to the type of plate and the type of assay being performed.

23. The apparatus of claim 18, wherein the means for positioning the sensed volume automatically positions the sensed volume after sensing the boundary interface.

24. The apparatus of claim 18, wherein the means for positioning the sensed volume automatically positions the sensed volume to maximize the signal detected from the composition.

25. The apparatus of claim 18, wherein the means for positioning the sensed volume adjusts the relative positions of the sensed volume and at least one boundary interface by moving the composition relative to portions of the means for transmitting light from the sensed volume.

26. The apparatus of claim 18, wherein the means for positioning the sensed volume adjusts the relative positions of the sensed volume and at least one boundary interface by moving portions of the means for transmitting light from the sensed volume relative to the composition.

27. The apparatus of claim 18, wherein the means for detecting light includes a detector selected from the group consisting of photomultiplier tubes (PMTs), photodiodes, avalanche photodiodes, charge-coupled devices (CCDs), and intensified CCDs.

28. An apparatus for detecting light transmitted from a sample, the apparatus comprising

a stage for supporting a sample in a microplate well, the sample having boundary interfaces defined by walls of the microplate well and an upper gas interface,

a light source positioned to deliver light to the sample,

a detector positioned to receive light transmitted from the sample, and

an optical relay structure that focuses a non-diffraction-limited spot into the sample without illuminating significantly any of the boundary interfaces of the sample.

29. The apparatus of claim 28, wherein the optical relay structure includes a confocal optics arrangement.

30. The apparatus of claim 28 further comprising a Z-axis optical adjustment device that automatically adjusts positions of the sensed volume relative to the boundary interfaces.

31. The apparatus of claim 30, wherein the Z-axis optical adjustment device automatically positions the sensed volume between the boundary interfaces according to stored parameters relating to the type of plate and the type of assay being performed.

32. The apparatus of claim 30, wherein the Z-axis optical adjustment device automatically positions the sensed volume after sensing the boundary interface.

33. The apparatus of claim 30, wherein the Z-axis optical adjustment device adjusts the relative positions of the sensed volume and at least one boundary interface by moving the composition relative to portions of the optical relay structure.

34. The apparatus of claim 30, wherein the Z-axis optical adjustment device adjusts the relative positions of the sensed volume and at least one boundary interface by moving portions of the optical relay structure relative to the composition.

35. The apparatus of claim 28, wherein the microplate well has a diameter D, the spot having a diameter of at least about 0.3D.

36. The apparatus of claim 28 further comprising

a sample handling mechanism capable of automatically detecting light serially from microplate wells in a density of at least about 384 wells per plate.

37. An apparatus for detecting light transmitted from a sample, the apparatus comprising

a stage for supporting a sample in a microplate well having a shape that is substantially frustoconical,

a light source positioned to deliver light to the sample,

a detector positioned to receive light transmitted from the sample, and

an optical relay structure located between the light source and the detector, the optical relay structure including confocal optics that create a sensed volume in the sample, at least a portion of the sensed volume having a shape conforming substantially to the shape of the well.

38. The apparatus of claim 37, wherein the sensed volume is characterized by a waist region having a diameter D and a Z-pick-up having a height of at least about 2D.

39. The apparatus of claim 37, wherein the microplate well has a diameter X, the sensed volume being character sized by a waist region having a diameter of at least about 0.3X.