Method and apparatus for delivery of submicroliter volumes onto a substrate

• US 20060024841A1
• Filed: 10/04/2005
• Published: 02/02/2006
• Est. Priority Date: 10/30/2000
• Status: Abandoned Application

• Current Assignee: Sequenom Incorporated (Laboratory Corporation Of America Holdings)
• Original Assignee: Sequenom Incorporated (Laboratory Corporation Of America Holdings)
• Inventors: Lin, Chao, Becker, Thomas, Heaney, Paul, Hanson, Aaron, Yao, Xian-Wei, Willis, Michael C.
• Application Number:
  US11/243,113
• Publication Number:
  US 20060024841A1
• Time in Patent Office: Days
• US Class Current: 436/180
• 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 2219/00387 Applications using probes
• CPC Class Codes: B01J 2219/00527 Sheets
• CPC Class Codes: B01J 2219/00529 DNA chips
• CPC Class Codes: B01J 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/0059 Sequential 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 2219/00608 DNA chips
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00619 using hydrophilic or hydrophobic regions
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00677 Ex-situ synthesis followed by deposition on the substrate
• CPC Class Codes: B01J 2219/00686 Automatic
• CPC Class Codes: B01J 2219/00689 using computers
• CPC Class Codes: B01J 2219/00691 using robots
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01J 2219/00725 Peptides
• CPC Class Codes: B01L 2200/0657 Pipetting powder
• CPC Class Codes: B01L 3/0244 using pins
• CPC Class Codes: B01L 3/0248 : Prongs, quill pen type dispenser
• CPC Class Codes: B82Y 30/00 : Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C40B 40/06 : Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: C40B 40/10 : Libraries containing peptides or polypeptides, or derivatives thereof
• CPC Class Codes: C40B 60/14 : for creating libraries
• CPC Class Codes: G01N 1/10 : in the liquid or fluent state burettes, pipettes B01L3/02; sampling of ground water E02D1/06; metering by volume of fluids or fluent solid material G01F11/00, G01F13/00
• CPC Class Codes: G01N 2035/00158 : Elements containing microarrays, i.e. "biochip"
• CPC Class Codes: G01N 2035/1037 : Using surface tension, e.g. pins or wires
• CPC Class Codes: G01N 35/0099 : comprising robots or similar manipulators robots per se B25J
• CPC Class Codes: G01N 35/028 : having reaction cells in the form of microtitration plates
• CPC Class Codes: G01N 35/10 : Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices G01N35/0099 takes precedence
• CPC Class Codes: G01N 35/1016 : Control of the volume dispensed or introduced
• CPC Class Codes: G01N 35/1065 : Multiple transfer devices
• CPC Class Codes: Y10T 436/11 : Automated chemical analysis
• CPC Class Codes: Y10T 436/112499 : with sample on test slide
• CPC Class Codes: Y10T 436/119163 : with aspirator of claimed structure
• CPC Class Codes: Y10T 436/24 : Nuclear magnetic resonance, electron spin resonance or other spin effects or mass spectrometry
• CPC Class Codes: Y10T 436/2575 : Volumetric liquid transfer

Abstract

A slotted pin tool, a delivery system containing the pin tool, a substrate for use in the system and methods using the pin tool and system are provided. The slotted pin tool contains a plurality of pins having slotted ends designed to fit around each loci of material deposited on a surface, such as a microarray, without contacting any of the deposited material. Sample is delivered by contacting the pin tool with the surface; the amount delivered is proportional to the velocity of the pin tool as it contacts the surface or the velocity of the liquid when movement of the pin is halted.

Claims

1. A method of delivering liquid samples to a substrate, the method comprising:

dipping a slotted pin tool having an open tip into a sample reservoir containing a liquid sample to be delivered onto the substrate, thereby drawing a volume of liquid sample up into the pin tool;

moving the slotted pin tool from the sample reservoir to an elevated position above the substrate such that the pin tool is above a target location that is to receive the liquid sample; and

lowering the slotted pin tool toward the substrate at a predetermined speed and then halting the movement of the pin tool toward the substrate, thereby expelling the liquid sample from the slotted pin tool onto the target location of the substrate.

2. A method of claim 1, wherein halting the movement of the slotted pin tool toward the substrate occurs substantially when the open tip of the slotted pin tool contacts the substrate.

3. A method of claim 2, wherein the open tip of the slotted pin tool is adapted to fit around material deposited at the target location on the substrate without making contact with any portion of the material.

4. A method of claim 1, wherein the slotted pin tool has a substantially cylindrical tip having a lateral slot forming a cavity that fits around a material at the target location on the substrate.

5. A method of claim 1, wherein the slotted pin tool has a substantially cylindrical tip having a lateral slot forming a cavity with a width of greater than approximately 75 μ

m.

6. A method of claim 5, wherein the cavity of the cylindrical tip has a width between about 100 μ

m up to approximately 300 μ

m.

7. A method of claim 5, wherein the cavity of the cylindrical tip has a width between about 30 μ

m up to approximately 1000 μ

m.

8. A method of claim 5, wherein the cavity of the cylindrical tip has a height greater than approximately 100 μ

m

9. A method of claim 1, wherein dipping the pin tool comprises dipping the tip into the reservoir, whereby liquid is drawn into the slot.

10. A method of claim 1, wherein dipping the pin tool comprises dipping the tip into the reservoir such that the sample fluid level has a depth at least equal to the height of the slot in the tip, and the dipping halts when the pin tool has been dipped to a depth equal to the pin tool slot.

11. A method of claim 1, wherein positioning of the slotted pin tool is effected by pattern recognition to determine correct positioning above the substrate and, then moving the slotted pin tool to the determined position.

12. A method of claim 1, wherein moving the slotted pin tool comprises identifying an orientation mark on the substrate that indicates an appropriate location of the substrate.

13. A method of claim 1, wherein lowering the slotted pin tool comprises moving the slotted pin tool at a predetermined speed of lowing.

14. A method of claim 1, further comprising changing the volume of liquid sample delivered onto the substrate by changing the speed of lowering.

15. A method of claim 14, wherein the speed of lowering is changed in response to known composition of the liquid sample.

16. A method of claim 1, further comprising:

cleaning the slotted pin tool in a liquid bath after a first liquid sample has been delivered; and

drying slotted pin tool prior to dipping the pin tool in the liquid sample.

17. A method of claim 16, wherein cleaning and drying occur after the first liquid sample has been delivered and before a second liquid sample is delivered.

18. A method of claim 16, wherein drying comprises moving an air flow air over the slotted pin tool.

19. A method of claim 16, wherein the liquid bath comprises an ultrasonic bath.

20. The method of claim 1, wherein the sample comprises matrix material for mass spectrometric analyses.

21. A method of claim 5, wherein the cavity of the cylindrical tip has a width between about 100 μ

m up to approximately 500 μ

m.

22. The method of claim 12, wherein the orientation mark comprises a symbology, or an electronic tag or a physical deformation of the substrate for identification of orientation and/or identification of the substrate from among a plurality thereof.

23. A system for delivery of liquid samples from one or more pin tools to target locations on a substrate, the system comprising:

a plurality of processing stations, each of which performs a procedure on the pin tool, wherein the pin tool includes a lower tip having a slot into which a liquid sample may be drawn;

a transport system that transports the slotted pin tool from processing station to processing station; and

a control system that loads the slotted pin tool by moving it to a loading station at which the slotted pin tool is loaded with the liquid sample, then moves the slotted pin tool to an elevated position above the substrate such that the pin tool is above a target location that is to receive the liquid sample, and then dispenses the liquid sample by lowering the slotted pin tool toward the substrate at a predetermined speed and then halts the movement of the pin tool toward the substrate, thereby expelling the liquid sample from the slotted pin tool onto the target location of the substrate.

24. A system of claim 23, wherein the control system halts the movement of the slotted pin tool toward the substrate substantially when the open tip of the slotted pin tool contacts the substrate.

25. A system of claim 24, wherein the open tip of the slotted pin tool is adapted to fit around a material at the target location on the substrate without making contact with any portion of the material.

26. A system of claim 23, wherein the slotted pin tool has a substantially cylindrical tip having a lateral slot forming a cavity that fits around a material at the target location on the substrate.

27. A system of claim 23, wherein the slotted pin tool has a substantially cylindrical tip having a lateral slot forming a cavity with a width of greater than approximately 100 μ

m.

28. A system of claim 27, wherein the cavity of the cylindrical tip has a width of approximately 300 μ

m.

29. A system of claim 27, wherein the cavity of the cylindrical tip has a height greater than approximately 100 μ

m.

30. A system of claim 23, wherein the control system loads the slotted pin tool by dipping the tip of the pin tool into a sample-containing reservoir at the loading station;

and the control system halts the dipping substantially when the pin tool has been dipped to a depth equal to the pin tool slot.

31. A system of claim 23, wherein the control system moves the slotted pin tool to the elevated position using pattern recognition techniques to determine correct positioning above the substrate.

32. A system of claim 23, wherein the control system moves the slotted pin tool to the elevated position by identifying an orientation mark on the substrate that indicates an appropriate location of the substrate.

33. A system of claim 23, wherein the control system lowers the slotted pin tool by moving the slotted pin tool at a predetermined speed of lowering.

34. A system of claim 33, wherein the control system changes the volume of liquid sample delivered onto the substrate by changing the speed of lowering.

35. A system of claim 33, wherein the control system changes the speed of lowering in accordance with known composition of the liquid sample.

36. A system of claim 23, wherein the control system further moves the slotted pin tool by cleaning the slotted pin tool in a liquid bath after a first liquid sample has been delivered and drying the slotted pin tool prior to dipping the pin tool in the liquid sample.

37. A system of claim 36, wherein the control system performs cleaning and drying after the first liquid sample has been delivered and before it delivers a second liquid sample.

38. A system of claim 36, wherein drying comprises moving an air flow air over the slotted pin tool.

39. A system of claim 36, wherein the liquid bath comprises an ultrasonic bath.

40. A pin tool for use in a sample delivery system, the pin tool comprising one or more slotted pins each having an open tip adapted to fit around a material at a target location on a substrate without making contact with any portion of the material, wherein:

the slotted pin tool is adapted to be dipped into a sample reservoir containing a liquid sample to be delivered onto the substrate, thereby drawing a volume of liquid sample up into the slotted pins in the pin tool.

41. A pin tool of claim 40, wherein a pin in the pin tool has a substantially cylindrical tip having a lateral slot forming a cavity with a width of greater than approximately 75 μ

m.

42. A pin tool of claim 41, wherein the cavity of the cylindrical tip has a width up to approximately 500 μ

m.

43. A pin tool of any of claims 40, wherein the cavity of the cylindrical tip has a height greater than approximately 100 μ

m.

44. A combination of a pin tool of claim 41 and a substrate comprising target loci for deposition of sample material, wherein the array of pins in the pin tool is matched to the array of loci on the substrate.

60. The pin tool of claim 40, wherein a pin in the tool is tapered.

45. A substrate for use in mass spectrometric analyses, comprising target locations defined by application of photoresist materials and photolithographic deposition;

wherein the resulting array of target locations on the substrate are less hydrophobic than the surrounding areas.

46. A substrate of claim 45, wherein a substrate starting surface is comprised of a material that has an available —

OH or primary amine.

47. The substrate of claim 45, wherein:

the substrate comprises two materials such that the one material has a contact angle that differs by at least about 20 degrees from the second material.

48. The substrate of claim 47, wherein the first material is polytetrafluoroethylene or a derivative thereof or is dimethyidichlorosilane (DMDCS);

and the second material is silicon or silicon dioxide, which form the target loci.

49. A combination of a pin tool and the substrate of claim 48, wherein the pin tool comprises at least one pin that is slotted.

55. A method of mass spectrometric analysis, comprising:

depositing matrix material and sample on the target loci of a substrate of claim 45;

introducing the substrate into a mass spectrometer for analysis of the samples; and

analyzing the samples by mass spectrometry.

56. The method of claim 55, wherein the sample comprises nucleic acids or proteins.

50. A combination comprising:

a substrate, comprising an array of target locations on a surfaces, wherein the target locations less hydrophobic than the surrounding areas; and

a pin tool comprising at least one pin having a substantially cylindrical tip with a lateral slot forming a cavity that fits around a material deposited at a target location on the substrate.

51. A combination of claim 50, wherein the pin has a substantially cylindrical tip having a lateral slot forming a cavity with a width of greater than approximately 75 μ

m, preferably greater than 100 μ

m.

52. A combination of claim 50, wherein the cavity of the cylindrical tip has a width between about 100 μ

m up to approximately 300 μ

m.

53. A combination of claim 50, wherein the cavity of the cylindrical tip in a pin has a height greater than approximately 100 μ

m.

54. A combination of claim 50, wherein:

the slotted pin tool is adapted to be dipped into a sample reservoir containing a liquid sample to be delivered onto the substrate, thereby drawing a volume of liquid sample up into the slotted pins in the pin tool, and then is moved to an elevated position above the substrate such that the pin tool is above the target location(s), and then is lowered toward the substrate such that upon halting the movement thereof the liquid sample is expelled from the slotted pin tool onto the target location of the substrate.

57. A method of preparing a substrate, comprising:

coating a surface of a substrate with a photoresist material and solidifying it on the surface;

exposing the surface through a mask to light of a wavelength that permits removal of the photoresist from exposed surface;

washing the photoresist off with a developer leaving an array of photoresist pads having on the surface;

baking the resulting substrate.

58. The method of claim 57, wherein the substrate is baked at between about 190-200°

C. for a period of about 50-70 minutes.

59. The method of claim 57, wherein the photoresist contains a diazonaphthoquinone sensitizer and a phenolic resin.