Microinjection chip, device, system and uses thereof for unicellular or multicellular organisms

Patent metadata

Field	Value
Publication number	US-2019388892-A1
Application number	US-201916449707-A
Country	US
Kind code	A1
Filing date	Jun 24, 2019
Priority date	Jun 24, 2018
Publication date	Dec 26, 2019
Grant date	—

Abstract

Disclosed herein are microinjection chips, devices, and systems for injection of unicellular or multicellular organisms. The microinjection chip and device disclosed herein include the microfluidic features, inlet port, pre-injection reservoir, injection channel and post injection channel in fluid communication with each other. The inlet port is adapted to sequentially move individual organisms into the injection channel, which is adapted to immobilize the individual organism in fluid. The injection channel features a side wall adapted to receive a microinjection pipette without a microinjection port and to reseal when the microinjection pipette is removed.

First claim

1 . A microinjection chip for injection of unicellular or multicellular organisms, comprising:
a) an inlet port; b) an injection channel, that does not comprise a microinjection port; and, c) a post injection reservoir, wherein the inlet port, the injection channel and the post injection reservoir are in fluid communication with each other; wherein the inlet port is adapted to sequentially move individual organisms into the injection channel, which is adapted by size and applied pressure to immobilize the individual organism in fluid, and the injection channel comprises a side wall adapted to receive a microinjection pipette without a microinjection port and reseal when the microinjection pipette is removed.

2 . The chip of claim 1 , wherein the side wall has a thickness from 10 μm to 40 μm.

3 . The chip of claim 1 , wherein the side wall is formed from a material that allows the microinjection pipette to enter and be removed without forming an opening, and without breaking or clogging the microinjection pipette.

4 . The chip of claim 3 , wherein the side wall comprises a silicone polymer.

5 . (canceled)

6 . The chip of claim 4 , wherein the silicone polymer comprises a polydimethylsiloxane (PDMS) elastomer.

7 . The chip of claim 1 , wherein the inlet port, the injection channel and the post injection reservoir are formed with a polymer material forming a chip comprising the inlet port, the injection channel and the post injection reservoir.

8 . The chip of claim 1 , further comprising a pre-injection reservoir connected in series to, and in fluid communication with, the inlet port and the injection channel.

9 . The chip of claim 1 , wherein the unicellular or multicellular organism is an embryo or oocyte.

10 . (canceled)

11 . (canceled)

12 . (canceled)

13 . A microinjection device for injection of unicellular or multicellular organisms, comprising:
a microinjection chip comprising:
a) an inlet port;
b) an injection channel, that does not comprise a microinjection port; and,
c) a post injection reservoir, wherein the inlet port, the injection channel and post injection reservoir are in fluid communication with each other; wherein the inlet port is adapted to sequentially move individual organisms into the injection channel, which is adapted to immobilize the individual organism in fluid, and the injection channel comprises a side wall adapted to receive a microinjection pipette without a microinjection port and reseal when the microinjection pipette is removed; and,
a backing bonded to the microinjection chip forming a bottom to the inlet port, injection channel and post injection reservoir.

14 . The device of claim 13 , wherein the backing comprises glass or plastic.

15 - 25 . (canceled)

26 . A microfluidic system for microinjection of unicellular or multicellular organisms, comprising:
a microinjection device comprising:
a) an inlet port;
b) an injection channel, that does not comprise a microinjection port; and,
c) a post injection reservoir, wherein the inlet port, the injection channel and the post injection reservoir are in fluid communication with each other; wherein the inlet port is adapted to sequentially move individual organisms into the injection channel, which is adapted to immobilize the individual organism in fluid, and the injection channel comprises a side wall adapted to receive the microinjection pipette without a microinjection port and reseal when the microinjection pipette is removed; and,
d) a backing bonded to the microinjection chip forming a bottom to the inlet port, the injection channel and the post injection reservoir; and,
a microinjection pipette.

27 - 37 . (canceled)

38 . A method for microinjection of a liquid material into a unicellular or multicellular organism, comprising:
a) adding a fluid comprising the unicellular or multicellular organism to the inlet port that is in fluid communication with an injection channel according to the device of claim 13 ; b) placing the device comprising the unicellular or multicellular organism onto a stage of a microscope that comprises a micromanipulator and microinjection pipette wherein the microinjection pipette is positioned in a resting position; c) injecting liquid material into the unicellular or multicellular organism by moving the microinjection pipette comprising the liquid material through the side wall and penetrating the unicellular or multicellular organism that is stabilized in fluid within the injection channel; and, d) moving the microinjection pipette to the resting position and moving the unicellular or multicellular organism to the post injection reservoir.

39 . The method of claim 38 , further comprising use of a syringe attached to the inlet port to move the unicellular or multicellular organism from the inlet port to the injection channel and into the post injection reservoir.

40 . The method of claim 38 , further comprising applying pressure to the inlet port to move the unicellular or multicellular organism from the inlet port to the injection channel and into the post injection reservoir.

41 . (canceled)

42 . The method of claim 38 , wherein the liquid material comprises nucleic acid, chemical compounds or enzymes in an aqueous solution.

43 . (canceled)

44 . The method of claim 38 , wherein the inlet port, the injection channel and the post injection reservoir are in fluid communication with each other and comprise an osmolarity controlled buffer solution.

45 . The method of claim 38 , wherein the unicellular or multicellular organism is an embryo or oocyte.

46 . The method of claim 38 , wherein the multicellular organism is a nematode or fruit fly larva.

47 . (canceled)

48 . (canceled)

49 . The method of claim 38 , wherein the injected multicellular organisms are removed from the post injection reservoir and selected by sorting fluorescently labeled recovered organisms and/or by selecting recovered non-fragmented organisms.

50 . A device for the fabrication of a microinjection chip of claim 1 comprising:
a) a bottom plate comprising a negative imprint of the inlet port, pre-injection reservoir, injection channel and a post-injection reservoir;
b) a top plate comprising:
i) a central opening in the shape of a footprint of the microinjection chip with nose shaped cut out wherein the depth of the central opening determines the height of the microinjection chip and the nose shaped cut out provides injection pipette access to the injection channel; and
ii) a curvilinear channel adjacent and parallel to the microinjection chip opening;

c) one or more screws used to align and connect the top plate to the bottom plate wherein optionally the screws are inserted through the plates in a perpendicular direction or into the edge of the bottom plate fitted with threaded holes;
d) a screw positioned perpendicular to the plates, located in the edge of the top plate and adjacent to the curvilinear channel, wherein the screw controls the distance between the nose-shaped cut-out and the microinjection chip opening and sets the thickness of the side wall.

51 . (canceled)

Assignees

• Nemametrix Inc
• Univ Oregon

Inventors

• Lockery Shawn

Classifications

• C12M35/00

  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 title

• B01L3/502761 Primary

  specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads or physically stretching molecules · CPC title

• B01L3/502715

  characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces · CPC title

• B01L2200/0668

  Trapping microscopic beads · CPC title

• B01L3/50273

  characterised by the means or forces applied to move the fluids · CPC title