Method and apparatus for incubation of a liquid reagent and target spots on a microarray substrate

• US 6,485,918 B1
• Filed: 07/02/2001
• Issued: 11/26/2002
• Est. Priority Date: 07/02/2001
• Status: Expired due to Fees

• Current Assignee: PerkinElmer LAS, Inc. (Perkinelmer Incorporated)
• Original Assignee: Packard Bioscience Co. (Massachusetts) (Revvity, Inc.)
• Inventors: St. Cyr, Paul, Regan, Donald, Lombardo, C. Todd, Milkowski, Robert, Schermer, Mack J.
• Primary Examiner(s): Chin, Christopher L.
• Assistant Examiner(s): COUNTS, GARY W
• Application Number:
  US09/897,621
• Time in Patent Office: 512 Days
• Field of Search: 422/58, 422/102, 422/104, 422/940, 422/942, 435/283.1, 435/6, 435/286.7, 435/299.2, 435/303.3, 435/305.3, 435/305.4, 435/7.1, 435/287.1, 435/99, 436/46, 436/174, 436/807, 436/518, 436/524, 366/267
• US Class Current: 506/43
• CPC Class Codes: B01F 31/31 using receptacles with deformable parts, e.g. membranes, to which a motion is imparted
• CPC Class Codes: B01F 33/30 Micromixers
• CPC Class Codes: B01F 33/40 Mixers using gas or liquid agitation, e.g. with air supply tubes
• CPC Class Codes: B01J 2219/00479 Means for mixing reactants or products in the reaction vessels
• CPC Class Codes: B01J 2219/00493 by sparging or bubbling with gases
• CPC Class Codes: B01J 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00605 the compounds being directly bound or immobilised to solid supports
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00621 by physical means, e.g. trenches, raised areas
• CPC Class Codes: B01J 2219/00626 Covalent
• CPC Class Codes: B01J 2219/00637 by coating it with another layer
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00702 Processes involving means for analysing and characterising the products
• CPC Class Codes: B01L 2300/0819 Microarrays; Biochips
• CPC Class Codes: B01L 3/50851 specially adapted for heating or cooling samples laboratory heating apparatus B01L7/00; incubators C12M
• CPC Class Codes: B01L 3/50853 with covers or lids
• CPC Class Codes: C40B 60/14 for creating libraries

Abstract

A method and apparatus for incubating a liquid reagent with target spots on a microarray substrate. A deformable cover is placed over the surface of the microarray substrate having the target spots with the liquid reagent between the microarray substrate and the deformable cover, and a device is used to apply a force to the deformable cover.

Claims

1. A method for incubating a liquid reagent with target spots on a first surface of a microarray substrate, comprising:

confining the liquid reagent in a space between a deformable cover having a gasket and the first surface of the microarray substrate;

applying at least first and second forces to the deformable cover, wherein the first and second forces compress the casket to seal the liquid reagent in the space; and

altering the magnitude of the first and second forces to agitate the liquid reagent while maintaining as substantially constant a value equal to the sum of the first force and the second force so that the gasket maintains the seal of the liquid reagent in the space.

2. The method of claim 1, wherein the deformable cover includes a semi-rigid material.

3. The method of claim 1, wherein the deformable cover includes the gasket and a lid.

4. The method of claim 1, wherein at least one air bubble exists in the space between the deformable cover and the first surface of the substrate.

7. The method of claim 1, further comprising varying a location on the deformable cover on which the first force is applied.

8. The method of claim 7, further comprising varying a location on the deformable cover on which the second force is applied.

9. The method of claim 1, wherein confining the liquid reagent includes:

placing the liquid reagent on the deformable cover; and

placing the first surface of the substrate having the target spots over the deformable cover such that the deformable cover surrounds the target spots.

10. The method of claim 9, wherein placing the liquid reagent on the deformable cover includes manually dispensing the liquid reagent.

11. The method of claim 9, wherein placing the liquid reagent on the deformable cover includes automatically dispensing the liquid reagent.

12. The method of claim 9, wherein placing the first side of the substrate over the deformable cover includes manually placing the substrate.

13. The method of claim 9, wherein placing the first side of the substrate over the deformable cover includes automatically placing the substrate.

14. The method of claim 1, further comprising controlling the temperature of the substrate.

15. The method of claim 14, wherein controlling the temperature includes placing a temperature control block over the substrate.

16. The method of claim 1, further comprising introducing an air bubble within the space.

17. The method of claim 16, wherein altering the magnitude of the first and second forces causes the air bubble to agitate the liquid reagent.

18. The method of claim 1, wherein applying at least first and second forces includes using rollers to apply the forces.

19. The method of claim 18, further comprising rolling the rollers over the deformable cover to alter locations on the deformable cover on which the forces are applied.

20. The method of claim 1, further comprising allowing at least one reaction between one of the target slots on the substrate and the liquid reagent to occur.

21. The method of claim 20, wherein the reaction is selected from the group consisting of hybridization, protein binding, immunoassays, and chemical binding.

22. The method of claim 20, further comprising at least partially removing the deformable cover from the substrate.

23. The method of claim 22, further comprising washing from the substrate the liquid reagent that has not reacted with the target spots.

24. The method of claim 23, further comprising drying the substrate.

5. An apparatus for incubating a liquid reagent with target spots on a first surface of a microarray substrate, comprising:

means for confining the liquid reagent in a space between a deformable cover having a gasket and the first surface of the substrate;

means for applying at least first and second forces to the deformable cover, wherein the first and second forces compress the gasket to seal the liquid reagent in the space; and

means for altering the magnitude of the first and second forces to agitate the liquid reagent while maintaining as substantially constant a value equal to the sum of the first force and the second force so that the gasket maintains the seal of the liquid reagent in the space.

6. The apparatus of claim 5, further comprising:

means for dispensing the liquid reagent in the space.

25. The apparatus of claim 5, further comprising a temperature control block having a surface to engage the substrate.

26. The apparatus of claim 5, further comprising a washer to wash the liquid reagent that has not reacted with the target spots from the substrate.

27. The apparatus of claim 26, further comprising a dryer to dry the substrate.

28. The apparatus of claim 5, wherein the deformable cover comprises a material selected from the group consisting of glass, polypropylene, polyethylene, Teflon silicone rubber, fluorosilcone, fluoroelastomer, or nitrile.