Side view imaging microwell array

• US 20070178012A1
• Filed: 07/18/2006
• Published: 08/02/2007
• Est. Priority Date: 07/19/2005
• Status: Abandoned Application

• Current Assignee: Physical Sciences Incorporated
• Original Assignee: Physical Sciences Incorporated
• Inventors: Ferguson, R. Daniel, Rosen, David, Ferrante, Anthony
• Application Number:
  US11/488,901
• Publication Number:
  US 20070178012A1
• Time in Patent Office: Days
• US Class Current: 422/82.50
• CPC Class Codes: B01L 2300/0654 Lenses; Optical fibres
• CPC Class Codes: B01L 2300/0829 Multi-well plates; Microtitration plates
• CPC Class Codes: B01L 2300/0851 Bottom walls
• CPC Class Codes: B01L 3/5085 for multiple samples, e.g. microtitration plates
• CPC Class Codes: B01L 9/523 for multisample carriers, e.g. used for microtitration plates
• CPC Class Codes: G01N 2021/0382 Frustoconical, tapered cell
• CPC Class Codes: G01N 21/0303 Optical path conditioning in cuvettes, e.g. windows; adapted optical elements or systems; path modifying or adjustment G01N21/031 - G01N21/15 take precedence
• CPC Class Codes: G01N 21/253 for batch operation, i.e. multisample apparatus analytical automats G01N35/00
• CPC Class Codes: G01N 21/55 Specular reflectivity
• 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 21/6458 Fluorescence microscopy fluorescence microscopes per se G02B21/0076 and G02B21/16
• CPC Class Codes: G01N 2201/0634 Diffuse illumination

Abstract

Methods and apparatus for imaging a sample using a microwell array are provided. The methods and apparatus allow side view imaging of a sample to acquire fluorescence or bright field images.

Claims

1. An apparatus for imaging a sample comprising

a substrate defining a well for holding the sample, the well comprising a bottom and a first wall;

and

a first optical surface adjacent the first wall of the well, the first optical surface directing radiation from the well to an imaging lens located below the substrate.

2. The apparatus of claim 1, wherein at least one wall of the well is adapted to orient a bottom surface of the sample adjacent a bottom plane of the well.

3. The apparatus of claim 1, wherein the first optical surface is adjacent two or more wells.

4. The apparatus of claim 1, further comprising a second optical surface adjacent a second wall of the well, the second optical surface adapted to direct incident radiation through said second wall to the sample.

5. The apparatus of claim 4, wherein the second optical surface is adjacent two or more wells.

6. The apparatus of claim 4, wherein the second optical surface directs the incident radiation to a side surface of the sample, and radiation reflected or emitted by the sample is directed to the imaging lens, thereby forming a side image of the sample.

7. The apparatus of claim 4, wherein at least one of the first and second optical surfaces comprises a prism formed in a lower portion of the substrate.

8. The apparatus of claim 1, wherein the first optical surface directs incident radiation traveling along a first optical path to a second, substantially orthogonal optical path to the sample through the wall of the well.

9. The apparatus of claim 1, further comprising an upper portion defining a tapered hole positionable over the well.

10. The apparatus of claim 9, wherein the upper portion comprises a surface adapted to diffuse an incident beam of radiation.

11. The apparatus of claim 1, wherein the sample is selected from the group consisting of an embryonic amphibian, a larval amphibian, an embryonic fish, a larval fish, and an adult fish,

12. The apparatus of claim 1, wherein the well comprises a rectangular cross-section.

13. The apparatus of claim 1, wherein the substrate defines an array of wells.

14. The apparatus of claim 1, wherein a cross section of the well is tapered.

15. The apparatus of claim 1, wherein the bottom of the well defines an area of about 1.5 millimeters wide and about 6 millimeters long.

16. The apparatus of claim 1, wherein the apparatus comprises 96 wells.

17. The apparatus of claim 1, having the dimensions of a standard 96 well microplate.

18. The apparatus of claim 17, wherein the apparatus is formed as a single piece.

23. A method of imaging a sample comprising depositing a sample into a well of the substrate of claim 1;

directing radiation into the well, and

receiving radiation reflected or emitted by the sample to form an image of the sample.

19. An apparatus for imaging a sample comprising:

a substrate defining a rectangular well for holding the sample, the well comprising a bottom, a first wall, and a second wall;

a first optical surface adjacent the first wall of the well, the first optical surface receiving incident radiation and directing the incident radiation through the first wall of the well;

a second optical surface adjacent the second wall of the well, the second optical surface receiving radiation from the well and direct the radiation from the well to an imaging lens located below the substrate, and

an upper portion of the substrate defining a tapered hole positionable over the well.

20. An apparatus for imaging a sample comprising:

a substrate defining a well for holding the sample;

means for directing incident radiation to the sample through a wall of the well; and

means for receiving radiation reflected or emitted by the sample to form an image of the sample.

21. The apparatus of claim 20, further comprising an upper portion, the upper portion defining a hole positionable over the well.

22. The apparatus of claim 21, wherein the upper portion comprises a surface adapted to diffuse an incident beam of radiation.

24. A method of imaging a sample comprising:

providing a substrate defining a well for holding the sample, the well comprising a bottom, a first wall, and a first optical surface, the first optical surface being adjacent the first wall and adapted to direct radiation from the well to an imaging lens located below the substrate;

directing incident radiation into the well, and

receiving radiation reflected or emitted by the sample into the imaging lens to form an image of the sample.

25. The method of claim 24, further comprising directing the incident radiation into the well through a second wall of the well using a second optical surface adjacent the second wall of the well.

26. The method of claim 24, wherein the first optical surface comprises a prism formed in the substrate.

27. The method of claim 24, further comprising facilitating introduction of the sample to the well using an upper portion of the substrate, the upper portion defining a tapered hole positionable over the well.

28. The method of claim 24, wherein at least one wall is adapted to orient a bottom surface of the sample adjacent the bottom of the well.

29. The method of claim 27, wherein the incident radiation comprises a beam of radiation, further comprising diffusing the incident beam of radiation prior to illuminating the sample.

30. The method of claim 24, further comprising acquiring a bright field image of the sample in the well.

31. The method of claim 24, further comprising acquiring a fluorescence image of the sample in the well.

32. The method of claim 24, wherein the incident radiation comprises fluorescence excitation light delivered to the sample through the top of the well.

33. The method of claim 24, wherein the sample is a zebra fish.

34. A method of imaging a sample comprising:

directing incident radiation to a sample in a well of a substrate;

receiving radiation reflected or emitted by the sample through a wall of the well, and

directing the reflected or emitted radiation via an optical surface formed in the substrate to an imaging lens to form an image of the sample.

35. An apparatus for imaging a sample comprising

a substrate defining a well for holding the sample, the well comprising a bottom and a first wall;

and

a first channel adjacent the first wall of the well, the first channel capable of receiving a turning optic capable of directing radiation from the well to an imaging lens located below the substrate.