Method and apparatus for parallel dispensing of defined volumes of solid particles

• US 20030124735A1
• Filed: 10/25/2002
• Published: 07/03/2003
• Est. Priority Date: 10/26/2001
• Status: Active Grant

• Current Assignee: Agena Bioscience Inc. (Mesa Laboratories Inc.), Grohmann Engineering GmbH (Tesla Inc.)
• Original Assignee: Sequenom Incorporated (Laboratory Corporation Of America Holdings)
• Inventors: Backes, Josef, Nanthakumar, Elizabeth, Unger, Siegfried
• Granted Patent: US 7,159,740 B2
• Time in Patent Office: Days
• US Class Current: 436/180
• CPC Class Codes: B01J 2219/00274 Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
• CPC Class Codes: B01J 2219/00315 Microtiter plates
• CPC Class Codes: B01J 2219/00364 Pipettes
• CPC Class Codes: B01J 2219/00369 in multiple or parallel arrangements
• CPC Class Codes: B01J 2219/00466 in a slurry
• CPC Class Codes: B01J 2219/005 Beads
• CPC Class Codes: B01L 2200/022 Variable spacings
• CPC Class Codes: B01L 2200/0657 Pipetting powder
• CPC Class Codes: B01L 2200/143 Quality control, feedback systems
• CPC Class Codes: B01L 2300/0829 Multi-well plates; Microtitration plates
• CPC Class Codes: B01L 2300/185 using a liquid as fluid
• CPC Class Codes: B01L 2400/086 using baffles or other fixed flow obstructions
• CPC Class Codes: B01L 3/0217 of the plunger pump type medical syringes A61M
• CPC Class Codes: B01L 3/0241 Drop counters; Drop formers making arrays for combinatorial libraries B01J19/0046; automation of dispensing for analysis G01N35/10
• CPC Class Codes: B01L 3/0244 using pins
• CPC Class Codes: B01L 7/02 Water baths; Sand baths; Air baths
• CPC Class Codes: B01L 7/52 with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples amplification or hybridisation processes per se C12Q1/68; controlling sequential reactions for synthesis B01J19/0046
• CPC Class Codes: B01L 9/56 Means for indicating position of a recipient or sample in an array
• CPC Class Codes: C40B 60/14 for creating libraries
• CPC Class Codes: G01N 2035/00168 Manufacturing or preparing test elements
• CPC Class Codes: G01N 2035/00207 : Handling bulk quantities of analyte
• CPC Class Codes: G01N 2035/00326 : Analysers with modular structure
• CPC Class Codes: G01N 2035/00346 : Heating or cooling arrangements
• CPC Class Codes: G01N 2035/00574 : Means for distributing beads
• CPC Class Codes: G01N 35/028 : having reaction cells in the form of microtitration plates
• CPC Class Codes: G01N 35/1011 : Control of the position or alignment of the transfer device
• CPC Class Codes: G01N 35/1065 : Multiple transfer devices
• CPC Class Codes: G01N 35/1074 : arranged in a two-dimensional array
• CPC Class Codes: Y10T 436/2575 : Volumetric liquid transfer

Abstract

A sample preparation system provides an automated process that dispenses appropriate amounts of solid material into wells of a microtiter plate for mixing into a suspension. The system utilizes an array of hollow tubes that are lowered into a particle reservoir such that particle material is forced up into the tubes. During loading, solid plungers in the tubes are positioned a known distance from the open end of the tubes. When the particle material is to be dispensed, the plungers are lowered to push the particle material out of the tubes and into receiving wells of the microtiter plate. The plunger stroke and internal diameter of the tubes defines the dispensed volume of particle material.

Claims

1. A method of operating a computer-controlled process line that dispenses particle material into locations of a solid support having multiple locations, the method comprising:

positioning an array of hollow tubes above a reservoir containing a bed of particle material;

engaging the tube array with the reservoir such that the lower ends of the tubes are pressed into the particle bed and a volume of particle material is forced into the tubes;

disengaging the tube array from the reservoir such that the volume of particle material is retained within each of the tubes, wherein the retained volume is defined by the tube from the lower end of each tube to a plunger within each tube;

positioning the array above the solid support such that the lower ends of the tubes are located above corresponding locations of the solid support; and

using the plungers to push the particle material retained within each tube out into the corresponding locations of the solid support.

2. A method as defined in claim 1, wherein the number of hollow tubes in the array is equal to the number of locations in the solid support.

3. A method as defined in claim 1, wherein the array of hollow tubes comprises 96 hollow tubes.

4. A method as defined in claim 3, wherein the hollow tubes in the array of hollow tubes are arranged in a grid of eight by twelve.

5. A method as defined in claim 1, wherein the array of hollow tubes comprises 384 hollow tubes.

6. A method as defined in claim 5, wherein the hollow tubes in the array of hollow tubes are arranged in a grid of sixteen by twenty-four.

7. A method as defined in claim 1, wherein the array of hollow tubes comprises 1536 hollow tubes.

8. A method as defined in claim 7, wherein the hollow tubes in the array of hollow tubes are arranged in grid of thirty-two by forty-eight.

9. A method as defined in claim 1, further comprising the step of packing the solid material within the reservoir by engaging the bed of particle material with a compacting cover.

10. A method as defined in claim 9, further comprising shaking the reservoir to flatten the bed of particle material.

11. A method as defined in claim 1, wherein the solid material comprises resin.

12. A method as defined in claim 9, wherein the step of packing the solid reagent comprises lifting the reservoir into the compacting cover until the bed of particle material is squeezed against an inside surface of the cover.

13. A method as defined in claim 1, wherein the plungers are connected to a plunger plate, and wherein the plungers are moved downward simultaneously using the plunger plate.

14. A method as defined in claim 1, wherein each of the hollow tubes in the array of hollow tubes is simultaneously cleaned by a skimming plate positioned between the array of hollow tubes and the reservoir, said skimming plate comprising a number of holes, each of said holes sized to slideably insert the hollow tubes therethrough.

15. A method as defined in claim 1, wherein the solid support comprises a microtiter plate and the locations comprise wells within said microtiter plate.

16. A method as defined in claim 1, wherein engaging the tube with the reservoir is performed by lowering the tube array until the lower ends of the tubes are pressed into the particle bed.

17. A method as defined in claim 1, wherein disengaging the tube array from the reservoir is performed by raising the tube array out of the reservoir.

18. A solid reagent dispensing station for filling locations of one or more solid supports with particles of solid reagent, said dispensing station comprising:

a dispenser assembly comprising;

a first plate;

a second plate;

an array of hollow tubes coupled to the first plate; and

an array of plungers coupled to the second plate, each of said plungers slideably inserted into the hollow tubes, wherein the second plate is configured to slide the plungers simultaneously up and down within the hollow tubes;

a reservoir configured to contain a bed of solid reagent particles, said reservoir sized to receive the array of hollow tubes;

a conveyor for transporting said one or more solid supports through the dispensing station; and

a transport mechanism connected to the dispenser assembly, the transport mechanism configured to transport the dispenser assembly between the reservoir and the one or more solid supports.

19. The solid reagent dispensing station of claim 18, wherein the number of hollow tubes in the array of hollow tubes equals the number of locations in one of the said one or more solid supports.

20. The solid reagent dispensing station of claim 18, wherein the array of hollow tubes comprises 96 hollow tubes.

21. The solid reagent dispensing station of claim 20, wherein the hollow tubes in the array of hollow tubes are arranged in a grid of eight by twelve.

22. The solid reagent dispensing station of claim 18, wherein the array of hollow tubes comprises 384 hollow tubes.

23. The solid reagent dispensing station of claim 22, wherein the hollow tubes in the array of hollow tubes are arranged in a grid of sixteen by twenty-four.

24. The solid reagent dispensing station of claim 18, wherein the array of hollow tubes comprises 1536 hollow tubes.

25. The solid reagent dispensing station of claim 24, wherein the hollow tubes in the array of hollow tubes are arranged in a grid of thirty-two by forty-eight.

26. The solid reagent dispensing station of claim 18, wherein the solid reagent comprises resin.

27. The solid reagent dispensing station of claim 18, wherein the station is an automated station controlled by a computer.

28. The solid reagent dispensing station of claim 18, wherein each of said hollow tubes has a proximal end with an opening, a distal end with an opening, and an inside surface, wherein each of said plungers comprises a proximal end and a distal end, and wherein a space is formed between the distal end of each of the plungers and the distal end of each of the hollow tubes, said space defining a predetermined volume sufficient for containing a predetermined amount of solid reagent.

29. The solid reagent dispensing station of claim 18, wherein the reservoir comprises:

a collar with an open proximal end, an open distal end, and an inner surface;

a base comprising an outer surface, said base shaped to sealably slide through the distal end of the collar, its outer surface in contact with the inner surface of the collar; and

a compacting lid removably engaged to the collar.

30. The solid reagent dispensing station of claim 29, wherein the compacting lid has a stick-resistant lower surface.

31. The solid reagent dispensing station of claim 18, further comprising a compacting lid having a flat lower surface, said compacting lid configured for lowering onto the reservoir and exerting pressure on the particles of reagent.

32. The solid reagent dispensing station of claim 31, wherein the lower surface of the compacting lid comprises a stick-resistant material.

33. The solid reagent dispensing station of claim 18, wherein the hollow tubes are cylindrical.

34. The solid reagent dispensing station of claim 18, further comprising a skimming plate suspended over the reservoir.

35. The solid reagent dispensing station of claim 34, wherein the skimming plate has a number of holes equal to the number of hollow tubes in the array, each of said holes sized to slideably insert the hollow tubes therethrough.

36. The solid reagent dispensing station of claim 18, wherein the solid support comprises a microtiter plate and the locations comprise wells within said microtiter plate.

37. A method of operating a computer-controlled process line that dispenses particle material into locations of a solid support having multiple locations, the method comprising:

positioning an array of hollow tubes over a skimming plate, wherein said skimming plate comprises a number of holes equal to the number of hollow tubes in the array, and wherein the hollow tubes and the holes are aligned;

positioning a reservoir comprising a bed of particle material underneath the skimming plate and the array of tubes;

lowering the array of hollow tubes through the holes in the skimming plate and into the reservoir, wherein the hollow tubes are forced into the bed of particle material;

lifting the array of hollow tubes until the tubes have withdrawn from the holes in the skimming plate, wherein particle material is lodged within each of said hollow tubes;

positioning the array of hollow tubes over a solid support, said solid support comprising a number of locations equal to the number of hollow tubes in the array, wherein the hollow tubes are lined up with the locations;

lowering the array of hollow tubes over the locations of the solid support; and

using plungers slideably inserted into each of the hollow tubes to simultaneously push the particle material out into the locations.

38. A method as defined in claim 37, wherein the solid support comprises a microtiter plate and the locations comprise wells within said microtiter plate.

39. A method as defined in claim 37, wherein the array of hollow tubes is coupled to one or more guide rails and is positioned over the skimming plate by sliding the array on said one or more guide rails along an axis Y from a point of origin.

40. A method as defined in claim 39, wherein the reservoir is coupled to one or more tracks and is positioned underneath the skimming plate by sliding the reservoir on said one or more tracks along an axis X, which is perpendicular to the axis Y.

41. A method as defined in claim 40, wherein the array of hollow tubes is positioned over the solid support by sliding the array on the one or more guide rails along the Y axis to said point of origin.

42. An apparatus for dispensing particle material into locations of a solid support having multiple locations, the apparatus comprising:

a first plate having a top surface and a bottom surface, said top surface having an array of holes leading into a corresponding array of hollow chambers;

an array of hollow tubes attached to the bottom surface of said first plate, said array of hollow tubes aligned with the holes and the hollow chambers;

a second plate having a top surface and a bottom surface;

an array of plungers attached to the bottom surface of said second plate, wherein the array of plungers and array of holes is aligned and the plungers of the array are inserted into the holes and extend into the hollow tubes;

one or more springs mounted in between the top surface of the first plate and the bottom surface of the second plate, forcing the two plates apart; and

one or more stop posts connecting the first plate to the second plate.

43. The apparatus of claim 42, wherein the number of hollow tubes is equal to the number of plungers.

44. The apparatus of claim 42, wherein the first and second plates are substantially flat.

45. The apparatus of claim 42, further comprising:

a third plate;

two lateral walls connecting the first flat plate to the third flat plate; and

an adjustment screw threaded through a hole in the third plate, said screw protruding through the bottom of the third plate and resting against the top of the second plate, said adjustment screw setting the second plate a predetermined maximum distance from the first plate.

46. The apparatus of claim 45, wherein the adjustment screw comprises markings representative of a volume of space within the hollow tubes.

47. The apparatus of claim 42, further comprising a computer controlled pneumatic pump in communication with the top surface of the second plate, said pump configured to apply a downward force on the second plate.

48. The apparatus of claim 42, wherein the solid support comprises a microtiter plate and the locations comprise wells within said microtiter plate.

49. A solid reagent dispensing station for filling locations of one or more solid supports with particles of solid reagent, said dispensing station comprising:

a reservoir configured to contain a bed of solid reagent particles;

a skimming plate having an array of holes, said skimming plate positioned above the reservoir; and

dispenser means for withdrawing a plurality of predetermined volumes of the solid reagent particles through the holes of the skimming plate and dispensing them in the locations of the one or more solid supports.

50. The solid reagent dispensing station of claim 49 further comprising a transport mechanism, wherein said transport mechanism comprises:

at least one position sensor for detecting the position of the dispenser means; and

a guide rail that is coupled to the dispenser means.

51. The solid reagent dispensing station of claim 49, wherein the reservoir comprises a hollow compacting lid and a vibrator plate inside the hollow lid.

52. The solid reagent dispensing station of claim 50, wherein the reservoir slideably rests on one or more tracks which are substantially perpendicular to the guide rail.

53. The solid reagent dispensing station of claim 49, wherein the solid support comprises a microtiter plate.

54. The solid reagent dispensing station of claim 49, wherein the dispenser means comprises an array of hollow tubes, each of which receives a plunger slideably inserted therein.

55. The solid reagent dispensing station of claim 54, wherein the number of holes in the array of holes is equal to the number of hollow tubes in the array of hollow tubes, and wherein the hollow tubes in the array of hollow tubes are arranged for complementary fit with the holes in the array of holes.

56. The solid reagent dispensing station of claim 49, wherein the reservoir comprises:

a collar with an open proximal end, an open distal end, and an inner surface;

a base comprising an outer surface, said base shaped to sealably slide through the distal end of the collar, its outer surface in contact with the inner surface of the collar; and

a compacting lid removably engaged to the collar.