Template methods and devices for preparing sample arrays

• US 20060046282A1
• Filed: 08/26/2004
• Published: 03/02/2006
• Est. Priority Date: 08/26/2004
• Status: Active Grant

• Current Assignee: United States Department Of Health And Human Services (Government of the United States of America)
• Original Assignee: The United States of America As Represented By The Secretary of Agriculture
• Inventors: Hewitt, Stephen M.
• Granted Patent: US 7,854,899 B2
• Time in Patent Office: Days
• US Class Current: 435/40.500
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01J 2219/00313 the reactor vessels being formed by arrays of wells in blocks
• CPC Class Codes: B01J 2219/00373 Hollow needles
• CPC Class Codes: B01J 2219/0043 for direct application of reagents, e.g. through openings in a shutter
• CPC Class Codes: B01J 2219/0074 Biological products
• CPC Class Codes: B01J 2219/00743 Cells
• CPC Class Codes: C40B 60/14 for creating libraries

Abstract

A method for preparing a microarray that includes placing at least one template over a first surface of the recipient block, wherein the template defines an array of openings and the recipient block has a plurality of receptacle holes, such that the array of openings are aligned with the plurality of receptacle holes. A needle or punch that contains a sample is inserted through the openings of the template. The sample then is inserted into the receptacle hole in the recipient block. A device is also disclosed that includes a platform defining (i) a first surface, and (ii) a first region configured to retain at least one recipient block; and a raised template defining an array of openings, secured to the first surface of the platform and positioned above the first region configured to retain the recipient block.

Claims

1. A method for preparing a microarray, comprising:

placing at least one template over a first surface of the recipient block, wherein the template defines an array of openings and the recipient block has a plurality of receptacle holes, such that the array of openings are aligned with the plurality of receptacle holes;

inserting a first needle or punch through the openings of the template, wherein the first needle or punch contains a sample; and

inserting the sample into the receptacle hole in the recipient block.

2. The method of claim 1, further comprising forming the receptacle holes in the recipient block by placing the template over the first surface of a recipient block;

and

inserting a second needle or punch through at least one opening in the template so as to bore a complementary receptacle hole in the recipient block.

3. The method of claim 1, further comprising securing the template to the recipient block.

4. The method of claim 3, wherein securing the template to the recipient block comprises adhesively adhering the template to the recipient block via a transparent adhesive tape.

5. The method of claim 2, wherein the boring of the receptacle holes in the recipient block results in an array of receptacle holes in the recipient block.

6. The method of claim 2, wherein the receptacle holes are cylindrical and are all of substantially the same depth.

7. The method of claim 2, wherein the first needle or punch is the same as the second needle or punch.

8. The method of claim 2, wherein the second needle or punch has at least one protrusion that has a dimension greater than the widest dimension of the opening of the template.

9. The method of claim 8, wherein the protrusion on the second needle or punch contacts the template so as to prevent the second needle or punch from being inserted any further into the recipient block.

10. The method of claim 1, wherein the first needle or punch has at least one protrusion that has a dimension greater than the widest dimension of the opening of the template.

11. The method of claim 10, wherein the protrusion on the first needle or punch contacts the template so as to prevent the first needle or punch from penetrating a plane of the first surface of the recipient block.

12. The method of claim 2, wherein the punch or needle withdraws a core from the recipient block to form the receptacle hole.

13. The method of claim 1, further comprising inserting the first needle or punch into a tissue donor block to obtain a tissue sample, introducing an end of the first needle or punch containing the tissue sample to one of the openings of the template, and then disgorging the tissue sample from the first needle or punch so as to introduce the tissue sample into the receptacle hole in the recipient block.

14. The method of claim 1, wherein the openings in the template are spaced substantially equally apart.

15. The method of claim 1, wherein the samples are all aligned substantially vertically within the receptacle holes, and the samples all occupy substantially the same depth in the receptacle holes.

16. The method of claim 6, wherein the cylindrical receptacle holes have a diameter of about 1.5 mm to about 4 mm, the openings of the template are circular and have a diameter of about 1.6 mm to about 4.1 mm, and the openings of the template are spaced apart from each other by about 1.5 mm to about 4 mm.

17. The method of claim 1, further comprising freezing the sample in the receptacle hole to produce a cryoarray.

18. The method of claim 1, further comprising maintaining the recipient block at or below freezing while inserting the sample into the receptacle hole in the recipient block to produce a cryoarray.

19. The method of claim 1, further comprising:

placing the recipient block on a base plate to produce a base plate/recipient block/template assembly;

placing the base plate/recipient block/template assembly in a cryostat; and

then

inserting the first needle or punch through the openings of the template to insert the sample into the receptacle hole in the recipient block.

20. The method of claim 1, wherein the recipient block comprises a frozen or freezable embedding medium.

21. A method of preparing an array of samples, comprising:

placing a first template over a first surface of a recipient block, wherein the first template defines a first array of first openings;

inserting a first punch or needle through at least one opening in the first template so as to bore a receptacle hole in the recipient block;

placing a second template over the first template, wherein the second template defines a second array of second openings, such that the first openings and the second openings are aligned axially;

inserting a second punch or needle through at least the second opening such that the second needle or punch does not penetrate a plane of the first surface of the recipient block, wherein the second punch or needle contains a sample; and

inserting the sample into the receptacle hole in the recipient block.

22. The method of claim 21, wherein the second punch or needle is also inserted through the first opening of the first template.

23. The method of claim 21, wherein the receptacle holes are cylindrical and are all of substantially the same depth.

24. The method of claim 21, wherein the first punch or needle has at least one protrusion that has a dimension greater than the widest distance of the first opening of the first template.

25. The method of claim 24, wherein the protrusion on the first punch or needle contacts the first template so as to prevent the first punch or needle and from being inserted any further into the recipient block.

26. The method of claim 24, wherein the second punch or needle has at least one protrusion that has a dimension greater than the widest dimension of the second opening of the second template.

27. The method of claim 26, wherein the protrusion on the needle or punch contacts the second template so as to prevent the second needle or punch from penetrating the plane of the first surface of the recipient block.

28. The method of claim 21, wherein the first openings in the first template and the second openings in the second template are each spaced substantially equally apart.

29. The method of claim 21, wherein the samples are all aligned substantially vertically within the receptacle holes, and the tissue samples all occupy substantially the same depth in the receptacle holes.

30. The method of claim 23, wherein the cylindrical receptacle holes have a diameter of about 1.5 mm to about 4 mm, the first openings of the first template and the second openings of the second template are each circular and have a diameter of about 1.6 mm to about 4.1 mm, and the first openings of the first template and the second openings of the second template are spaced apart from each other by about 1.5 mm to about 4.0 mm.

31. The method of claim 21, wherein the recipient block comprises a frozen or freezable embedding medium.

32. The method of claim 21, wherein the first punch or needle is the same as the second punch or needle.

33. A construct for preparing a sample microarray, comprising:

a recipient block defining an array of receptacle holes; and

at least one template attached to a first surface of the recipient block, and defining an array of openings that are complementary with the array of receptacle holes in the recipient block.

34. The construct of claim 33, comprising a first template attached to the first surface of the recipient block and a second template attached to the first template.

35. A device for preparing an array of samples, comprising:

a platform defining (i) a first surface, and (ii) a first region configured to retain at least one recipient block; and

a raised template defining an array of openings, secured to the first surface of the platform and positioned above the first region configured to retain the recipient block.

36. The device of claim 35, wherein the first region configured to retain a recipient block defines a first cavity configured to receive the recipient block.

37. The device of claim 36, wherein the raised template and the first cavity form a slot adapted to receive the recipient block.

38. The device of claim 35, wherein the openings of the template are circular and have a diameter of about 1.5 mm to about 4.0 mm, and the openings of the template are spaced apart from each other by about 1.5 mm to about 4.0 mm.