Array-type multiple cell injector

• US 5,262,128 A
• Filed: 10/23/1989
• Issued: 11/16/1993
• Est. Priority Date: 10/23/1989
• Status: Expired due to Term

• Current Assignee: National Institutes of Health (NIH) (Government of the United States of America)
• Original Assignee: United States Department Of Health And Human Services (Government of the United States of America)
• Inventors: Brownstein, Michael J., Leighton, Stephen B.
• Primary Examiner(s): Housel, James C.
• Assistant Examiner(s): Torres, Ramon
• Application Number:
  US07/425,254
• Time in Patent Office: 1,485 Days
• Field of Search: 422/100, 422/102, 436/180, 436/165, 436/63, 436/809, 435/293, 435/294, 435/301, 435/287, 73/863.31, 73/864.72, 935/53, 935/85
• US Class Current: 422/522
• CPC Class Codes: B01L 2300/0829 Multi-well plates; Microtitration plates
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 3/0244 using pins
• CPC Class Codes: B01L 3/0248 Prongs, quill pen type dispenser
• CPC Class Codes: B01L 3/5085 for multiple samples, e.g. microtitration plates
• CPC Class Codes: C12M 23/12 Well or multiwell plates C12M25/04 takes precedence
• CPC Class Codes: C12M 23/50 Means for positioning or orientating the apparatus C12M41/08 takes precedence
• CPC Class Codes: C12M 29/00 Means for introduction, extraction or recirculation of materials, e.g. pumps pumps per se F04B
• CPC Class Codes: C12M 35/00 Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion machines for extracting juice from animal or plant tissue by electroplasmolysis A23N1/006, processes employing electric or wave energy B01J19/08; treatment of microorganisms or enzymes with electrical or wave energy C12N13/00; methods for cell fusion C12N15/02; introduction of foreign genetic material C12N15/87
• CPC Class Codes: C12N 15/87 Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
• CPC Class Codes: G01N 2035/00237 Handling microquantities of analyte, e.g. microvalves, capillary networks
• CPC Class Codes: Y10S 436/809 Multifield plates or multicontainer arrays
• CPC Class Codes: Y10T 436/2575 Volumetric liquid transfer

Abstract

An injection device injects small amounts of injectate into each of a plurality of cells. A first plate, which may be made from a silicon chip, includes a large number of evenly spaced cell wells, in a regular array. Each well includes a through hole in the chip. A top plate, also formed from a silicon chip, includes a plurality of injection needles, arranged in a regular array corresponding to the position of the cell wells. A suspension of cells is washed over the first plate and individual cells are retained by the individual wells. A vacuum may be applied to the cell wells through a manifold beneath the first plate, thus to retain the cells in place. Excess cells and suspension are removed by washing the plate. The second plate, including the injection needles, is positioned on the first plate so that the needles pierce the plurality of cells retained in the wells, thus simultaneously introducing the injectate into a large number of individual cells.

Claims

1. Apparatus for injection of a medium into a plurality of cells, comprising:

first means having a plurality of cell wells arranged in a regular array for holding a plurality of individual cells in a regular array;

second means having a plurality of injection needles spaced in an array corresponding to said regular array of said cell wells for injecting the medium into the individual cells; and

third means for positioning said first and second means accurately with respect to one another.

2. Apparatus for injection of a medium into a plurality of cells as recited in claim 1, wherein said regular array is a linear array.

5. Apparatus for injection of a medium into a plurality of cells as recited in claim 2, wherein said wells are cylindrical.

6. Apparatus for injection of a medium into a plurality of cells as recited in claim 5, wherein said wells include a counter-sunk conical upper section.

7. Apparatus for injection of a medium into a plurality of cells as recited in claim 1, wherein said first means comprises a chip and said wells comprise through holes having openings on opposing surfaces of said chip.

8. Apparatus for injection of a medium into a plurality of cells as recited in claim 7, further comprising pressure means for providing a pressure differential to retain the cells in said wells.

9. Apparatus for injection of a medium into a plurality of cells as recited in claim 8, wherein said pressure means comprises a manifold chamber in sealing engagement with one surface of said chip and wherein said second means is positioned on the opposite surface of said chip.

10. Apparatus for injection of a medium into a plurality of cells as recited in claim 1, wherein said injection needles are substantially solid.

11. Apparatus for injection of a medium into a plurality of cells as recited in claim 1, wherein said injection needles each comprise a hollow portion.

12. Apparatus for injection of a medium into a plurality of cells as recited in claim 11, wherein said hollow portion of said injection needles includes a cavity at a tip portion thereof contacting one of the cells.

13. Apparatus for injection of a medium into a plurality of cells as recited in claim 11, wherein said second means comprises a chip and said injection needles project from one surface thereof to said first means, each of said needles including a through hole passing therethrough and through said chip.

14. Apparatus for injection of a medium into a plurality of cells as recited in claim 13, further comprising a manifold on a surface of said chip opposite to said one surface, said manifold holding the medium to be injected by said needles.

15. Apparatus for injection of a medium into a plurality of cells as recited in claim 14, further comprising pressure means for pressurizing said manifold to inject the medium.

16. Apparatus for injection of a medium into a plurality of cells as recited in claim 1, wherein said first and second means each comprises a substantially planar chip, said planar chips arranged substantially parallel to one another, and said third means comprises means for moving said planar chips towards and away from one another, to provide contact between each injection needle and a cell in each said well.

3. Apparatus for injection of a medium into a plurality of cells, comprising:

first means having a plurality of cell wells arranged in a regular array for holding a plurality of cells in a regular array;

second means having a plurality of injection needles spaced in an array corresponding to said regular array of said cell wells; and

third means for positioning said first and second means accurately with respect to one another, wherein

each of said wells has a predetermined size selected to match a size of a single cell, thereby limiting each well to hold therein only one cell.

4. Apparatus for injection of a medium into a plurality of cells as recited in claim 3, wherein said regular array is a two-dimensional array.

17. Apparatus for injection of a medium into a plurality of cells, comprising:

first means having a plurality of cell wells arranged in a regular array for holding a plurality of individual cells in a regular array;

second means having a plurality of injection needles spaced in an array corresponding to said regular array of said cell wells for injecting the medium into the individual cells;

third means for positioning said first and second means accurately with respect to one another;

said first and second means each comprising a substantially planar chip, said planar chips arranged substantially parallel to one another, and said third means comprising means for moving said planar chips towards and away from one another, to provide contact between each injection needle and a cell in each said well, wherein

said means for moving comprises a flexibly hinged structure having a pair of arms in a tweezer-like arrangement.

18. Apparatus for injection of a medium into a plurality of cells as recited in claim 17, wherein said means for moving comprises electrically controlled motion generating means.

19. Apparatus for injection of a medium into a plurality of cells as recited in claim 18, wherein said electrically controlled motion generating means comprises a stepper motor.

20. Apparatus for injection of a medium into a plurality of cells, comprising:

first means having a plurality of cell wells arranged in a regular array for holding a plurality of individual cells in a regular array;

second means having a plurality of injection needles spaced in an array corresponding to said regular array of said cell wells for injecting the medium into the individual cells; and

third means for positioning said first and second means accurately with respect to one another, wherein

said first and second means each comprises a plate, at least one of said plates having a convex curvature towards the other, thereby to facilitate contact between said needles and cells in said wells by a rolling motion of one plate relative to the other.

21. Apparatus for injection of a medium into a plurality of cells, comprising:

first means having a plurality of cell wells for holding a plurality of cells in a regular array;

second means having an injection needle; and

third means for positioning said first and second means accurately with respect to one another for injecting the medium into each of the cells in the array.

22. Apparatus for injection of a medium into a plurality of cells as recited in claim 21, wherein each of said cell wells has a predetermined size selected to match a size of a single cell, thereby limiting each well to accept only a single cell therein, and said second means comprises a plurality of injection needles corresponding to the plurality of cell wells for individually injecting the medium into each single cell.

26. Apparatus for injection of a medium into a plurality of cells as recited in claim 21, wherein:

each cell well in said first means has a predetermined size selected to match a size of a single cell, thereby limiting each well to accept only a single cell therein,

said second means comprises only a single injection needle and

said third means moves said single injection needle to individual positions respectively corresponding to the positions of individual ones of said plurality of cell wells in said regular array for individually injecting the single cells in said regular array of cell wells.

27. Apparatus for injection of a medium into a plurality of cells as recited in claim 21, wherein:

each cell well in said first means has a predetermined size with a diameter less than approximately 1 mm and selected to match a size of a single cell, thereby limiting each well to accept only a single cell therein, and

said second means comprises a plurality of injection needles arranged in a linear array for injecting the medium into each of a linear array of single cells.

23. Apparatus for injection of a medium into a plurality of cells, comprising:

first means having a plurality of cell wells positioned in a regular array for holding a plurality of individual cells, each cell well having a size selected to match a size of a single cell,

second means having at least one injection needle spaced apart from said regular array of said cell wells for injecting the medium into an individual cell in one of said regular array of cell wells; and

third means for successively positioning said first and second means with respect to one another to position said at least one injection needle in successive positions corresponding to said regular array of cell wells for successively individually injecting each of the cells.

24. Apparatus for injection of a medium into a plurality of cells as recited in claim 23, wherein said second means comprises only a single injection needle and said third means moves said single injection needle to individual positions respectively corresponding to the positions of individual ones of said plurality of cell wells in said regular array for individually injecting the single cells in said regular array of cell wells.

25. Apparatus for injection of a medium into a plurality of cells as recited in claim 23, wherein said second means comprises a plurality of injection needles arranged in a linear array for injecting the medium into each of a linear array of single cells.