Systems and methods for preparing low volume analyte array elements

• US 6,024,925 A
• Filed: 01/23/1997
• Issued: 02/15/2000
• Est. Priority Date: 01/23/1997
• Status: Expired due to Term

• Current Assignee: Agena Bioscience Inc. (Mesa Laboratories Inc.)
• Original Assignee: Sequenom Incorporated (Laboratory Corporation Of America Holdings)
• Inventors: Little, Daniel P., Koster, Hubert
• Primary Examiner(s): McKane, Elizabeth
• Assistant Examiner(s): Moazzam, Fariborz
• Application Number:
  US08/787,639
• Time in Patent Office: 1,118 Days
• Field of Search: 422/99, 422/100, 422/65, 222/394, 73/863.32, 73/864.15, 73/864.22, 73/864.24, 73/864.25
• US Class Current: 422/507
• 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 2219/00315 Microtiter plates
• CPC Class Codes: B01J 2219/00317 Microwell devices, i.e. having large numbers of wells
• CPC Class Codes: B01J 2219/00387 Applications using probes
• CPC Class Codes: B01J 2219/00468 by manipulation of individual beads
• CPC Class Codes: B01J 2219/00497 Features relating to the solid phase supports
• CPC Class Codes: B01J 2219/005 Beads
• CPC Class Codes: B01J 2219/00504 Pins
• CPC Class Codes: B01J 2219/00511 Walls of reactor vessels
• CPC Class Codes: B01J 2219/0052 in the shape of elongated tubes
• CPC Class Codes: B01J 2219/00527 Sheets
• 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 2219/0061 The surface being organic
• 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 2219/00648 by the use of solid beads
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/0072 Organic compounds
• CPC Class Codes: B01J 2219/00722 : Nucleotides
• CPC Class Codes: B01J 2219/00725 : Peptides
• CPC Class Codes: B01L 3/0262 : using touch-off at substrate or container
• CPC Class Codes: B01L 3/0268 : using pulse dispensing or spraying, eg. inkjet type, piezo actuated ejection of droplets from capillaries
• CPC Class Codes: C07B 2200/11 : Compounds covalently bound to a solid support
• CPC Class Codes: C07F 9/2408 : of hydroxyalkyl compounds
• CPC Class Codes: C07F 9/2429 : of arylalkanols
• CPC Class Codes: C07H 21/00 : Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• 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 2035/00237 : Handling microquantities of analyte, e.g. microvalves, capillary networks
• CPC Class Codes: G01N 2035/1037 : Using surface tension, e.g. pins or wires
• CPC Class Codes: G01N 2035/1069 : by adjusting the spacing between multiple probes of a single transferring head
• CPC Class Codes: G01N 35/1067 : for transfer to or from containers having different spacing
• CPC Class Codes: G01N 35/1074 : arranged in a two-dimensional array
• CPC Class Codes: Y10T 436/25 : including sample preparation
• CPC Class Codes: Y10T 436/25125 : Digestion or removing interfering materials
• CPC Class Codes: Y10T 436/2575 : Volumetric liquid transfer

Abstract

The invention provides, in one aspect, serial and parallel dispensing tools that can deliver defined and controlled volumes of fluid to generate multi-element arrays of sample material on a substrate surface. The substrates surfaces can be flat or geometrically altered to include wells of receiving material. In one embodiment, the invention provides a tool that allows the parallel development of a sample array. To this end, the tool can be understood as an assembly of vesicle elements, or pins, wherein each of the pins can include a narrow interior chamber suitable for holding manual liter volumes of fluid. Each of the pins can fit inside a housing that forms an interior chamber. The interior chamber can be connected to a pressure source that will control the pressure within the interior chamber to regulate the flow of fluid within the interior chamber of the pins. In a further embodiment, the invention then passes the prepared sample arrays to a plate assembly that disposes the sample arrays for analysis by mass spectrometry.

Claims

1. A dispensing apparatus for dispensing nanoliter volumes of fluid in chemical or biological procedures onto the surface of a substrate, comprising:

a housing having a plurality of sides and a bottom portion having formed therein a plurality of apertures, wherein the sides and bottom portion of said housing define an interior volume,

one or more fluid transmitting vesicles, mounted within said apertures, and having a fluid holding chamber of a size for holding and dispensing nanoliter volumes of fluid, said fluid holding chamber being disposed in fluid communication with said interior volume of said housing, and

dispensing means in communication with said interior volume of said housing for selectively dispensing nanoliter volumes of fluid from said nanoliter volume sized fluid transmitting vesicles when fluid holding chambers of said vesicles are fully loaded, whereby said dispensing means dispenses nanoliter volumes of the fluid onto the surface of the substrate when the apparatus is disposed over and in registration with the surface of the substrate.

2. The apparatus of claim 1, wherein each said fluid transmitting vesicle has an open proximal end and a distal tip portion that extends beyond said housing bottom portion when mounted within said apertures, said open proximal end disposing said fluid holding chamber in fluid communication with said interior volume when mounted with said apertures.

3. The apparatus of claim 1, wherein said one or more fluid transmitting vesicles are removably and replaceably mounted within said apertures of said housing.

4. The apparatus of claim 1, wherein said one or more fluid transmitting vesicles include a glue seal for fixedly mounting said vesicles within said housing.

5. The apparatus of claim 1, wherein said fluid holding chamber includes a narrow bore dimensionally adapted for being filled with the fluid through capillary action.

6. The apparatus of claim 1, wherein each said fluid holding chamber of said one or more fluid transmitting vesicles are sized to cause said fluid holding chamber to be substantially filled with the fluid through capillary action.

7. The apparatus of claim 1, comprising a plurality of fluid transmitting vesicles, wherein said plurality of fluid transmitting vesicles comprise an array of fluid delivering needles.

8. The apparatus of claim 7, wherein said fluid delivering needles are fabricated from metal.

9. The apparatus of claim 7, wherein said fluid delivering needles are fabricated from glass.

10. The apparatus of claim 7, wherein said fluid delivering needles are fabricated from silica.

11. The apparatus of claim 7, wherein said fluid delivering needles are fabricated from polymeric material.

12. The apparatus of claim 1, wherein the number of said one or more fluid transmitting vesicles is less than or equal to the number of wells of a multi-well substrate.

13. The apparatus of claim 1, comprising a plurality of fluid transmitting vesicles, an further comprising mechanical biasing means of mechanically biasing said plurality of fluid transmitting vesicles into sealing contact with said housing bottom portion, wherein the house further comprises a top portion.

14. The apparatus of claim 13, wherein each of the fluid transmitting vesicles has a proximal end portion that includes a flange, and further comprising a sealer element disposed between the flange and an inner surface of the housing bottom portion for forming a seal between the interior volume and an external environment.

15. The apparatus of claim 14, wherein said mechanical biasing means includes a plurality of spring elements each of which are coupled at one end to said proximal end of each said one or more fluid transmitting vesicles, and at another end to an inner surface of said housing top portion, said spring element applying a mechanical biasing force to said vesicle proximal end to form said seal.

16. The apparatus of claim 1, wherein said housing further includes a top portion, and further comprising securing means for securing said housing top portion to said housing bottom portion.

17. The apparatus of claim 16, wherein said securing means comprises a plurality of fastener-receiving apertures formed within one of said top and bottom portions of said housing, and a plurality of fasteners for mounting within said apertures for securing together said housing top and bottom portions.

18. The apparatus of claim 1, wherein said dispensing means comprises a pressure source fluidly coupled to said interior volume of said housing for disposing said interior volume at a selected pressure condition.

19. The apparatus of claim 18, wherein said fluid transmitting vesicles are filled through capillary action, and wherein said dispensing means further comprises means for varying said pressure source to dispose said interior volume of said housing at varying pressure conditions, said means for varying disposing said interior volume at a selected pressure condition sufficient to offset said capillary action to fill the fluid holding chamber of each vesicle to a predetermined height corresponding to a predetermined fluid amount.

20. The apparatus of claim 19, wherein said means for varying further comprises fluid selection means for selectively discharging a selected nanoliter volume fluid amount from said chamber of each said vesicle.

21. The apparatus of claim 1, wherein said fluid transmitting vesicle has a proximal end that opens onto said interior volume of sid housing, and wherein said fluid holding chamber of said vesicles are sized to cause said fluid holding chamber to be substantially filled with the fluid through capillary action without forming a meniscus at said proximal open end.

22. The apparatus of claim 1, wherein said dispensing means comprises fluid selection means for selectively varying the amount of fluid dispensed from said fluid holding chamber of each vesicle.

23. The apparatus according to claim 1, comprising a plurality of vesicles, wherein a first portion thereof includes fluid holding chambers of a first size and a second portion includes fluid holding chambers of a second size, whereby a plurality of fluid volumes can be dispensed.

24. The apparatus of claim 22, wherein said fluid selection means comprises a pressure source coupled to said housing and in communication with said interior volume for disposing said interior volume at a selected pressure condition, and

adjustment means coupled to said pressure source for varying said pressure within said interior volume of said housing to apply a positive pressure in said fluid chamber of each said fluid transmitting vesicle to vary the amount of fluid dispensed therefrom.

25. The apparatus of claim 1, comprising a plurality of fluid transmitting vesicles.

26. A system, comprising the apparatus of claim 1, and further comprising:

a robotic arm in attached to the housing, wherein the robotic arm is for holding and moving the housing to insert the vesicles into a fluid source for filling the vesicles with a nanoliter volume of fluid and for disposing said housing comprising said vesicles adjacent to the substrate to allow for dispensing fluid from the vesicles onto the surface of a substrate adjacent thereto.

27. The system of claim 26, further comprising a central processing means for processing data and for executing program instructions to provide information for controlling the movement and operation of the robotic arm.

28. The system of claim 26, wherein the substrate is for performing mass spectrometry on samples deposited thereon.

29. A fluid dispensing apparatus for dispensing nanoliter volumes of a fluid in chemical or biological procedures into one or more wells of a multi-well substrate, comprising

a housing having a plurality of sides and a bottom portion having formed therein a plurality of apertures, said sides and bottom portion defining an interior volume,

a plurality of fluid transmitting vesicles, each vesicle for holding a nanoliter volume of a fluid, mounted within said apertures having a fluid holding chamber disposed in communication with said interior volume of said housing,

a fluid volume selection and dispensing means in communication with said interior volume of said housing for variably selecting an amount of the fluid loaded into said fluid holding chambers of said vesicles to be dispensed from a single set of plurality of fluid transmitting vesicles, and

whereby said dispensing means dispenses a selected amount of the fluid into the wells of the multi-well substrate when the apparatus is disposed over and in registration with the substrate, wherein the selected amount of fluid is a nanoliter volume.

30. The fluid dispensing apparatus of claim 29, wherein said fluid volume selection and dispensing means is adapted to select various amounts of fluid to be dispensed from said single set of vesicles.

31. The fluid dispensing apparatus of claim 29, wherein said fluid volume selection and dispensing means comprises a pressure source fluidly coupled to said interior volume of said housing for dispensing said interior volume at a selected pressure condition.

32. The fluid dispensing apparatus of claim 31, further comprising means for varying the pressure within the interior volume of the housing to select the amount of fluid to dispense from said fluid transmitting vesicles.

33. The fluid dispensing apparatus of claim 31, wherein said fluid transmitting vesicles are filled with the fluid through capillary action, and further comprising means for varying said pressure source to dispose said interior volume of said housing at varying pressure conditions, said means for varying dispensing said interior volume at a pressure condition sufficient to offset said capillary action to fill the fluid holding chamber of each vesicle to a predetermined height corresponding to a predetermined fluid amount.

34. The fluid dispensing apparatus of claim 29, wherein said fluid selection means comprises

a pressure source coupled to said housing and in communication with said interior volume for dispensing said interior volume at a selected pressure condition, and

adjustment means coupled to said pressure source for varying said pressure within said interior volume of said housing to apply a positive pressure in said fluid chamber of each said fluid transmitting vesicle to vary the amount of fluid dispensed therefrom.

35. A fluid dispensing apparatus for dispensing fluid in chemical or biological procedures into one or more wells of a multi-well substrate, said apparatus comprising

a housing having a plurality of sides and top and bottom portions, said bottom portion having formed therein a plurality of apertures, said sides and top and bottom portions of said housing defining an interior volume,

a plurality of fluid transmitting vesicles, mounted within said apertures, each vesicle having a fluid holding chamber sized to hold a nanoliter volume of the fluid, said fluid holding chamber being disposed in fluid communication with said volume of said housing and

mechanical biasing means for mechanically biasing said plurality of said transmitting vesicles into sealing contact with said housing bottom portion, wherein the apparatus dispenses nanoliter volumes of fluid.

36. The fluid dispensing apparatus of claim 35, wherein each said fluid transmitting vesicle has a proximal end portion that includes a flange, and further comprising a sealer element disposed between the flange and an inner surface of the housing bottom portion for forming a pressure and fluid seal between the interior volume an external environment.

37. The fluid dispensing apparatus of claim 35, wherein said mechanical biasing means includes a plurality of spring elements each of which are coupled at one end to said proximal end of said fluid transmitting vesicle, and at another end to an inner surface of said housing top portion, said spring elements applying a mechanical biasing force to said vesicle proximal end to form said fluid and pressure seal.

38. The fluid dispensing apparatus of claim 35, further comprising securing means for securing said housing top portion to said housing bottom portion.

39. The fluid dispensing apparatus of claim 38, wherein said securing means comprises a plurality of fastener-receiving apertures formed within one of said top and bottom portions of said housing, and a plurality of fasteners for mounting within said apertures for securing said housing top and bottom portions together.

40. The fluid dispensing apparatus of claim 35, further comprising dispensing means in communication with said interior volume of said housing for selectively dispensing the fluid from said fluid transmitting vesicles when the fluid is loaded within said fluid holding chambers of said vesicles, whereby said dispensing means dispenses the fluid into the wells of the multi-well substrate when the apparatus is disposed over and in registration with the substrate.

41. The fluid dispensing apparatus of claim 40, wherein said dispensing means comprises a pressure source fluidly coupled to said interior volume of said housing for dispensing said interior volume at a selected pressure condition.

42. The fluid dispensing apparatus of claim 35, wherein said plurality of fluid transmitting vesicles are removably and replaceably mounted within said apertures of said housing.

43. The fluid dispensing apparatus of claim 35, wherein said plurality of fluid transmitting vesicles comprise an array of fluid delivering needles.

44. The fluid dispensing apparatus of claim 40, wherein said fluid transmitting vesicles are filled with the fluid through capillary action, and wherein said dispensing means further comprises means for varying said pressure source to dispense said interior volume of said housing at varying pressure conditions, said means for varying disposing said interior volumes at a selected pressure condition sufficient to offset said capillary action to fill the fluid holding chamber of each vesicle to a predetermined height corresponding to a predetermined fluid amount.

45. The fluid dispensing apparatus of claim 40, wherein said dispensing means comprises fluid selection means for selectively varying the amount of fluid dispensed from said fluid holding chamber of each vesicle.

46. The fluid dispensing apparatus of claim 35, further comprising a pressure source coupled to the housing and in communication with the interior volume for dispensing the interior volume at a selected pressure condition, and

adjustment means coupled to the pressure source for varying the pressure within the interior volume of the housing to apply a positive pressure to the fluid chamber of each the fluid transmitting vesicle to vary the amount of fluid dispensed therefrom.

47. A system for dispensing nanoliter volumes of fluid in chemical or biological procedures onto the surface of a substrate, comprising:

a pin assembly having one or more apertures and one or more fluid transmitting pins, mounted within the apertures, wherein the pins comprise a solid shaft of material, wherein one end is for capturing a nanoliter volume of fluid thereon; and

a robotic arm in contact with the pin assembly, wherein the robotic arm is for holding and moving the assembly to insert the end of the pins into a fluid source for capturing fluid on the end of the pins or to dispose the pin assembly adjacent to the surface of a substrate to dispense fluid onto the surface of a substrate by contacting the fluid on the end of the pins with the surface of the substrate.

48. The apparatus of claim 47, further comprising a central processing means for processing data and for executing program instructions to provide information for controlling the movement and operation of the robotic arm.

49. The apparatus of claim 47, wherein the end of the pins for capturing a nanoliter volume of fluid comprises a shape selected from the group consisting of flat, star-shaped, concave, pointed solid, pointed semi-hollow, and angled on one or both sides.

50. The apparatus of claim 47, wherein the pin assembly comprises a plurality of pins in an array.

51. A system for dispensing nanoliter volumes of fluid in chemical or biological procedures onto the surface of a substrate, comprising:

an assembly, comprising a capillary element for containing a volume of fluid to be dispensed through an orifice of the capillary and a transducer element in contact with the capillary for effecting physical deformation of the capillary causing ejectment of a nanoliter volume of fluid from the capillary;

a robotic arm for holding an assembly and for aligning the assembly adjacent to the surface of the substrate to dispense nanoliter volumes of fluid from the capillary to the surface of the substrate aligned therewith; and

mounting means for securing the robotic arm to the assembly.

52. The system of claim 51, further comprising a central processing means for processing data and for executing program instructions to provide information for controlling the movement and operation of the robotic arm.

53. The system claim 51, wherein the transducer element is selected from the group consisting of a piezoelectric, electric, electrorestrictive, magnetorestrictive, and electromechanical transducer.