Bead dispensing system

• US 6,887,431 B1
• Filed: 02/15/2000
• Issued: 05/03/2005
• Est. Priority Date: 02/16/1999
• Status: Expired due to Fees

• Current Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Original Assignee: Applera Corporation (Thermo Fisher Scientific Incorporated), Applied Biosystems Group (Thermo Fisher Scientific Incorporated)
• Inventors: Lehto, Dennis, Vann, Charles S.
• Primary Examiner(s): Warden, Jill
• Assistant Examiner(s): Gordon, Brian R.
• Application Number:
  US09/506,870
• Time in Patent Office: 1,904 Days
• Field of Search: 422/68.1, 422 63- 65, 422/81, 422 99-100, 422/102, 422/131, 430/528, 430531-532, 430/43, 430/49, 430/54, 430177-178, 430/801, 4352572-2573, 4352884-2887, 221/264, 221/95
• US Class Current: 506/43
• 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 2219/00283 Reactor vessels with top opening
• CPC Class Codes: B01J 2219/00308 interchangeably mounted in racks or blocks
• 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 2219/00369 in multiple or parallel arrangements
• CPC Class Codes: B01J 2219/00468 by manipulation of individual beads
• CPC Class Codes: B01J 2219/005 Beads
• CPC Class Codes: B01J 2219/00545 Colours
• CPC Class Codes: B01J 2219/00549 2-dimensional
• CPC Class Codes: B01J 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00648 by the use of solid beads
• CPC Class Codes: B01J 2219/0065 by the use of liquid beads
• 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 2219/00689 using computers
• CPC Class Codes: B01J 2219/00691 using robots
• CPC Class Codes: B01J 2219/00693 Means for quality control
• CPC Class Codes: B01J 2219/00702 Processes involving means for analysing and characterising the products
• CPC Class Codes: B01J 2219/00722 : Nucleotides
• CPC Class Codes: B01L 2200/021 : Adjust spacings in an array of wells, pipettes or holders, format transfer between arrays of different size or geometry
• CPC Class Codes: B01L 2200/0647 : Handling flowable solids, e.g. microscopic beads, cells, particles
• CPC Class Codes: B01L 2200/0657 : Pipetting powder
• CPC Class Codes: B01L 2200/143 : Quality control, feedback systems
• CPC Class Codes: B01L 2300/0654 : Lenses; Optical fibres
• CPC Class Codes: B01L 2300/0864 : comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2400/0415 : electrical forces, e.g. electrokinetic
• CPC Class Codes: B01L 2400/043 : magnetic forces
• CPC Class Codes: B01L 2400/0487 : fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/049 : vacuum
• CPC Class Codes: B01L 3/021 : Pipettes, i.e. with only one conduit for withdrawing and redistributing liquids
• CPC Class Codes: B01L 3/0289 : Apparatus for withdrawing or distributing predetermined quantities of fluid B01L3/02 takes precedence; sample taking G01N1/00; sample taking within automatic analysers G01N35/00; volume measuring in general G01F
• CPC Class Codes: B01L 3/563 : Joints or fittings in general F16L; Separable fluid transfer means to transfer fluids between at least two containers, e.g. connectors
• 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 60/14 : for creating libraries
• CPC Class Codes: C40B 70/00 : Tags or labels specially adapted for combinatorial chemistry or libraries, e.g. fluorescent tags or bar codes
• CPC Class Codes: G01N 2035/00574 : Means for distributing beads
• 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 35/1065 : Multiple transfer devices
• CPC Class Codes: Y10T 436/11 : Automated chemical analysis
• CPC Class Codes: Y10T 436/255 : including use of a solid sorbent, semipermeable membrane, or liquid extraction
• CPC Class Codes: Y10T 436/2575 : Volumetric liquid transfer

Abstract

A bead dispensing system is provided for delivering small amounts of substances onto substrates. The system can include, for example, a movable support structure having an array of spaced-apart projections depending from its lower side. An attraction source, such as a vacuum, magnetic, and/or electrostatic force, is operable at each projection end region to attract and retain one bead. The projection array can be aligned with an array of bead-receiving regions of a substrate, e.g., an array of spaced-apart wells of a micro-plate or card. In one embodiment, a plurality of reagent-carrying beads are picked up, retained at respective projection end regions, and moved to a location over a multi-well plate. The beads are then released in a fashion permitting each bead to land in a respective well. The system of the invention is particularly useful for fabricating arrays of reagents.

Claims

1. A system for picking up a plurality of submillimeter beads from a bead supply and transferring said beads to a desired location, comprising:

a plurality of projections depending from a support at spaced-apart locations defining an array;

a cavity formed at a lower end region of each of said projections, each of said cavities defined by (i) a lower opening, (ii) an upper ceiling region, and (iii) a sidewall extending between said lower opening and said upper ceiling region;

wherein said upper ceiling region defines a surface extending inwardly from said sidewall;

wherein each cavity has a substantially constant diameter along a region extending between its lower opening and its upper ceiling region, such that lines extending longitudinally along confronting inner surfaces of each sidewall are substantially parallel to one another; and

an attraction source, operable at said projection end regions, effective to draw beads from said supply into said cavities and to releasably retain said beads therein.

2. The system of claim 1, wherein at least a lower portion of said sidewall of each projection comprises a resiliently flexible material, such that said flexible lower portion of said sidewall can bend while maintaining the shape of said cavity so that said lower opening can bend to face said beads in said bead supply.

3. The system of claim 1, wherein the diameter of said lower opening is between about 100-1,250 micrometers and the longitudinal length of said sidewall, from said lower opening and to said upper ceiling, is between about 0.50-1.25 times the diameter of said lower opening.

4. The system of claim 1, wherein the diameter of said lower opening is between about 250-750 micrometers and the longitudinal length of said sidewall, from said lower opening and to said upper ceiling, is between about 0.75-1.10 times the diameter of said lower opening.

5. The system of claim 1, wherein said cavity is dimensioned to receive at least half of one submillimeter bead, and to preclude entry therein of a substantial portion of a second such bead.

6. The system of claim 2, wherein said resiliently flexible sidewall has a generally cylindrical shape, with both an inner diameter and a longitudinal depth of between about 100-1,250 micrometers.

7. The system of claim 6, wherein said inner diameter and longitudinal depth are between about 350-425 micrometers.

8. The system of claim 1, wherein said cavity is formed by a resiliently flexible, tubular sleeve fit over the end of a respective projection, said sleeve having an overhang region extending below a terminal end of said projection defining said sidewall;

and wherein said terminal end of said projection, facing said cavity, defines said upper ceiling region of said cavity.

9. A system for picking up a plurality of submillimeter beads from a bead supply and transferring said beads to a desired location, comprising:

a plurality of projections depending from a support at spaced-apart locations defining an array;

a cavity formed at a lower end region of each of said projections, each of said cavities defined by (i) a lower opening, (ii) an upper ceiling region, and (iii) a sidewall extending between said lower opening and said upper ceiling region;

wherein said upper ceiling region defines a surface extending inwardly from said sidewall;

an attraction source, operable at said projection end regions, effective to draw beads from said supply into said cavities and to releasably retain said beads therein;

a plurality of ampules for containing said bead supply, said ampules disposed in an array alignable with said projection array; and

a plurality of covers, each cover configured to extend over an upper opening of one of said ampules.

10. The system of claim 9, wherein each of said covers is a frangible membrane.

11. The system of claim 9, wherein said support is held by a frame that is

(i) adapted to pivot about a generally vertical axis, rendering movable said projection array along a generally arcuate or circular pathway, and

(ii) adapted for reciprocal linear motion along a generally vertical pathway;

such movement permitting said projections to be aligned with said ampule array and lowered so that each projection can enter a respective one of said ampules.

12. The system of claim 9, wherein one of said ampules holds a plurality of submillimeter beads that carry a first set of analyte-specific reagents, and another of said ampules holds a plurality of submillimeter beads that carry a second set of analyte-specific reagents;

said first and second reagent sets differing from each other by at least one analyte-specific component.

13. A system for picking up a plurality of submillimeter beads from a bead supply and transferring said beads to a desired location, comprising:

a plurality of projections depending from a support at spaced-apart locations defining an array;

a cavity formed at a lower end region of each of said projections, each of said cavities defined by (i) a lower opening, (ii) an upper ceiling region, and (iii) a sidewall extending between said lower opening and said upper ceiling region;

wherein said upper ceiling region defines a surface extending inwardly from said sidewall; and

an attraction source, operable at said projection end regions, effective to draw beads from said supply into said cavities and to releasably retain said beads therein;

wherein each of said projections is a capillary tube having an axial lumen extending therethrough, each lumen having (i) a first end that opens into a respective one of said cavities through said ceiling region, and (ii) a second end disposed in fluid communication with a pressure-control assembly;

wherein said pressure-control assembly includes (i) a vacuum pump operable to establish a reduced pressure within each of said lumens, said reduced pressure defining said attraction source; and

(ii) a pump operable to establish an increased pressure within each of said lumens, said increased pressure effective to displace any beads retained in said cavities.

14. The system of claim 13, wherein each of said lumens has an inner diameter at said first end that is smaller than the diameter of a respective cavity at a location directly adjacent said ceiling region.

15. The system of claim 14, wherein each of said cavities has an inner diameter at a location directly adjacent said ceiling region of greater than 275 micrometers, and each of said lumens has an inner diameter at said first end of between 100-275 micrometers.

16. A system for picking up a plurality of submillimeter beads from a bead supply and transferring said beads to a desired location, comprising:

a plurality of projections depending from a support at spaced-apart locations defining an array;

a cavity formed at a lower end region of each of said projections, each of said cavities defined by (i) a lower opening, (ii) an upper ceiling region, and (iii) a sidewall extending between said lower opening and said upper ceiling region;

wherein said upper ceiling region defines a surface extending inwardly from said sidewall;

an attraction source, operable at said projection end regions, effective to draw beads from said supply into said cavities and to releasably retain said beads therein; and

a detection system having a field of view extending along each of said projection end regions, and adapted to sense the presence or absence of a bead retained in said cavities;

wherein said detection system includes a plurality of elongated light-conductive fibers, each fiber having one end that extends along one of said projections and faces said cavity, and a second end disposed in optical communication with a camera device.

18. The system of claim 16, wherein the center-to-center pitch of the second-opening array is reduced by a factor of at least about 3, as compared to that of the first-opening array.

19. The system of claim 16, further comprising a substrate, wherein said substrate is a micro-card having a plurality of wells disposed in an array alignable under said second-opening array.

20. The system of claim 18, further comprising

a detection system having a field of view extending into each of said conduits, and adapted to sense the presence or absence of a bead in each well of said microcard.

22. The system of claim 20, wherein said detection system includes a pair of image capture devices.

23. The system of claim 20, wherein the captured and displayed image further provides an indication of whether more than one bead is retained in the lower end region cavity of any projection.

17. A system for picking up a plurality of submillimeter beads from a bead supply and transferring said beads to a desired location, comprising:

a plurality of projections depending from a support at spaced-apart locations defining an array;

a cavity formed at a lower end region of each of said projections, each of said cavities defined by (i) a lower opening, (ii) an upper ceiling region, and (iii) a sidewall extending between said lower opening and said upper ceiling region;

wherein said upper ceiling region defines a surface extending inwardly from said sidewall;

an attraction source, operable at said projection end regions, effective to draw beads from said supply into said cavities and to releasably retain said beads therein; and

a conduit assembly having a plurality of conduits for separately channeling a plurality of submillimeter beads released from said cavities to desired locations on a substrate, said conduits having (i) first openings at their upper ends disposed in an array having a center-to-center pitch like that of the projection array such that the first openings are substantially alienable under the projections, and (ii) second openings at their lower ends disposed in an array having a center-to-center pitch like that of the substrate array such that the second openings are substantially aligned thereover;

wherein said second openings are disposed in an array having a center-to-center pitch substantially smaller than that of the first-opening array.

21. A system for picking up a plurality of submillimeter beads from a bead supply and transferring said beads to a desired location, comprising:

a plurality of projections depending from a support at spaced-apart locations defining an array;

a cavity formed at a lower end region of each of said projections, each of said cavities defined by (i) a lower opening, (ii) an upper ceiling region, and (iii) a sidewall extending between said lower opening and said upper ceiling region;

wherein said upper ceiling region defines a surface extending inwardly from said sidewall;

an attraction source, operable at said projection end regions, effective to draw beads from said supply into said cavities and to releasably retain said beads therein; and

a detection system having a field of view extending along each of said projection end regions, and adapted to sense the presence or absence of a bead retained in said cavities;

wherein said detection system includes at least one image capture device positioned to capture and display an image that includes the lower end region of each of said projections to provide an indication of the presence or absence of a bead retained in the lower end region cavity of each projection.

24. A system for picking up a plurality of submillimeter beads from a bead supply and transferring said beads to a desired location, comprising:

a plurality of projections depending from a support at spaced-apart locations defining an array;

a cavity formed at a lower end region of each of said projections, each of said cavities defined by (i) a lower opening, (ii) an upper ceiling region, and (iii) a sidewall extending between said lower opening and said upper ceiling region;

wherein said upper ceiling region defines a surface extending inwardly from said sidewall;

an attraction source, operable at said projection end regions, effective to draw beads from said supply into said cavities and to releasably retain said beads therein; and

a conduit assembly having a plurality of conduits for separately channeling a plurality of submillimeter beads released from said cavities to desired locations on a substrate, said conduits having (i) first openings at their upper ends disposed in an array having a center-to-center pitch like that of the projection array such that the first openings are substantially alignable under the projections, and (ii) second openings at their lower ends disposed in an array having a center-to-center pitch like that of the substrate array such that the second openings are substantially aligned thereover;

wherein each of said conduits is curved along a longitudinal direction from said first opening to said second opening.

26. The system of claim 24, further comprising:

(i) a carousel adapted for rotation about a central axis, said carousel (a) pivotally supporting said parallelogram linkage assembly for movement radially about a central axis of rotation of the carousel, and (b) having a substrate holding area adjacent to said parallelogram linkage assembly; and

(ii) a stationary rail extending along an inner region of said carousel and having a continuous bearing surface in mechanical communication with said parallelogram linkage assembly, said bearing surface having (a) a region disposed a first distance from central axis and at a first vertical height, whereat said conduit array assumes said lowered position over said substrate holding area, and (b) a region disposed a second distance from said central axis, shorter than said first distance, and at a second vertical height, higher than said vertical height, whereat said conduit array assumes said raised position.

25. A system for picking up a plurality of submillimeter beads from a bead supply and transferring said beads to a desired location, comprising:

a plurality of projections depending from a support at spaced-apart locations defining an array;

a cavity formed at a lower end region of each of said projections, each of said cavities defined by (i) a lower opening, (ii) an upper ceiling region, and (iii) a sidewall extending between said lower opening and said upper ceiling region;

wherein said upper ceiling region defines a surface extending inwardly from said sidewall;

an attraction source, operable at said projection end regions, effective to draw beads from said supply into said cavities and to releasably retain said beads therein;

a conduit assembly having a plurality of conduits for separately channeling a plurality of submillimeter beads released from said cavities to desired locations on a substrate, said conduits having (i) first openings at their upper ends disposed in an array having a center-to-center pitch like that of the projection array such that the first openings are substantially alienable under the projections, and (ii) second openings at their lower ends disposed in an array having a center-to-center pitch like that of the substrate array such that the second openings are substantially aligned thereover; and

a parallelogram linkage assembly supporting said conduit assembly for reciprocal arcuate movement between a raised position and a lowered position.

27. A system for picking up a plurality of submillimeter beads from a bead supply and transferring said beads to a desired location, comprising:

a plurality of projections depending from a support at spaced-apart locations defining an array;

a cavity formed at a lower end region of each of said projections, each of said cavities defined by (i) a lower opening, (ii) an upper ceiling region, and (iii) a sidewall extending between said lower opening and said upper ceiling region;

wherein said upper ceiling region defines a surface extending inwardly from said sidewall;

an attraction source, operable at said projection end regions, effective to draw beads from said supply into said cavities and to releasably retain said beads therein; and

a conduit assemble having a plurality of conduits for separately channeling a plurality of submillimeter beads released from said cavities to desired locations on a substrate, said conduits having (i) first openings at their upper ends disposed in an array having a center-to-center center pitch like that of the projection array such that the first openings are substantially alignable under the projections, and (ii) second openings at their lower ends disposed in an array having a center-to-center pitch like that of the substrate array such that the second openings are substantially aligned thereover; and

a substrate, wherein said substrate is provided with a pair of spaced-apart indexing holes, each being aligned with a respective indexing pin depending from a lower side of said conduit array;

whereupon registering said indexing pins in said indexing holes substantially aligns said second-opening array with said well array of said substrate.

28. A system for picking up a plurality of submillimeter beads from a bead supply and transferring said beads to a desired location, comprising:

a plurality of projections depending from a support at spaced-apart locations defining an array;

a cavity formed at a lower end region of each of said projections, each of said cavities defined by (i) a lower opening, (ii) an upper ceiling region, and (iii) a sidewall extending between said lower opening and said upper ceiling region;

wherein said upper ceiling region defines a surface extending inwardly from said sidewall;

an attraction source, operable at said projection end regions, effective to draw beads from said supply into said cavities and to releasably retain said beads therein;

a conduit assembly having a plurality of conduits for separately channeling a plurality of submillimeter beads released from said cavities to desired locations on a substrate, said conduits having (i) first openings at their upper ends disposed in an array having a center-to-center pitch like that of the projection array such that the first openings are substantially alignable under the projections, and (ii) second openings at their lower ends disposed in an array having a center-to-center pitch like that of the substrate array such that the second openings are substantially aligned thereover;

a detection system having a field of view extending into each of said conduits, and adapted to sense the presence or absence of a bead on said substrate under each of said second openings;

wherein said detection system further comprises;

a radiation source adapted to illuminate said substrate at locations below each of said second openings; and

a plurality of elongated light-conductive fibers, each fiber having (i) one end disposed to receive light traveling up through a respective conduit, and (ii) a second end that communicates with a camera device.

30. The system of claim 28, wherein said capillary-size region of each conduit has an inner diameter of less than about 1 mm.

31. The system of claim 28, wherein capillary-size region of each conduit has inner sidewalls that are hydrophilic.

33. The system of claim 31, wherein said cover system further comprises:

an idler reel for holding said cover material in said supply position;

a driven reel for taking up said cover material in said take-up position;

a movable piston for effecting said linear reciprocal motion, said movable piston having a lower face on which said movable blade is mounted; and

a resiliently compliant, substantially planar surface provided on said lower face along a region between said movable blade for pressing said cover material against an upper surface of said substrate.

34. The system of claim 31, wherein said cover material comprises an optically clear film.

29. A system for picking up a plurality of submillimeter beads from a bead supply and transferring said beads to a desired location, comprising:

a plurality of projections depending from a support at spaced-apart locations defining an array;

a cavity formed at a lower end region of each of said projections, each of said cavities defined by (i) a lower opening, (ii) an upper ceiling region, and (iii) a sidewall extending between said lower opening and said upper ceiling region;

wherein said upper ceiling region defines a surface extending inwardly from said sidewall;

an attraction source, operable at said projection end regions, effective to draw beads from said supply into said cavities and to releasably retain said beads therein;

a conduit assembly having a plurality of conduits for separately channeling a plurality of submillimeter beads released from said cavities to desired locations on a substrate, said conduits having (i) first openings at their upper ends disposed in an array having a center-to-center pitch like that of the projection away such that the first openings are substantially alignable under the projections, and (ii) second openings at their lower ends disposed in an array having a center-to-center pitch like that of the substrate array such that the second openings are substantially aligned thereover; and

a support structure, wherein said first openings are disposed in an array along one side of said support structure, and said second openings are disposed in an array along an opposite side of said support structure, wherein said first-opening array is arranged with a center-to-center pitch larger than that of said second-opening array, and wherein a region of each conduit extending from a respective one of said second openings is of capillary size, such that a liquid placed in contact with said second-opening array can be drawn at least partially into said conduits by way of capillary action.

32. A system for picking up a plurality of submillimeter beads from a bead supply and transferring said beads to a desired location, comprising:

a plurality of projections depending from a support at spaced-apart locations defining an array;

a cavity formed at a lower end region of each of said projections, each of said cavities defined by (i) a lower opening, (ii) an upper ceiling region, and (iii) a sidewall extending between said lower opening and said upper ceiling region;

wherein said upper ceiling region defines a surface extending inwardly from said sidewall;

an attraction source, operable at said projection end regions, effective to draw beads from said supply into said cavities and to releasably retain said beads therein;

a covering system for covering said beads after said beads have been disposed at said desired locations on a substrate, wherein said desired locations comprise an array of wells formed in said substrate, said covering system comprising;

a continuous web of a cover material mounted for movement from a supply position to a take-up position; and

a shearing blade mounted for reciprocal linear motion along a direction substantially normal to said web for cutting a portion of said cover material at a region between said supply position and said take-up position.