IP Library Granted Patent US 10,233,419
Granted Patent B2
US 10,233,419 · App. 15/199,549 · Granted Mar 19, 2019

Apparatuses and methods for electroporation

Inventors: John Cesarek (Redwood City, CA); Travis Lee (Burlingame, CA); Richard Hansen (San Carlos, CA)
Assignee: Zymergen Inc.
C12M35/02C12N15/87
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Quick Facts
Patent No.
US 10,233,419
App. No.
15/199,549
Granted
Mar 19, 2019
Kind
B2
Abstract

Disclosed are apparatuses, systems, and methods for performing electroporation.

Claims (60)

1. An apparatus comprising:

a plurality of cuvette holders, wherein each cuvette holder is configured to hold a plurality of cuvettes and includes a bottom having a thermal conductivity greater than 150 W/(m K), a sidewall having a thermal conductivity greater than 150 W/(m K), a plurality of cuvette positioning features, a first electrode, and a plurality of second electrodes that are electrically isolated from each other and from the first electrode, wherein:

the sidewall and the bottom are in thermal connection with each other and at least partially define an internal volume of the cuvette holder,

each cuvette positioning feature is configured to restrain a movement of a cuvette that is inserted into the internal volume such that a portion of each cuvette that is inserted into the internal volume is in thermal connection with the sidewall,

the first electrode and the plurality of second electrodes are configured such that insertion of a cuvette into the internal volume causes the cuvette to be electrically coupled to the first electrode and to one of the plurality of second electrodes; and

a plate having a thermal conductivity greater than 150 W/(m K) and including a first surface and a plurality alignment features, wherein:

each alignment feature is configured to restrain a movement of a cuvette holder that is positioned on the first surface with respect to the first surface, and

the bottom of each cuvette holder that is placed on the first surface and restrained by at least one alignment feature is in thermal connection with the first surface; and

a fluid flowpath in thermal connection with the first surface.

2. The apparatus of claim 1 , wherein:

each cuvette holder further includes a second sidewall having a thermal conductivity greater than 150 W/(m K),

the second sidewall is in thermal connection with the bottom, and

each cuvette positioning feature is further configured to restrain a movement of a cuvette that is inserted into the internal volume such that a second portion of each cuvette that is inserted into the internal volume is in thermal connection with the second sidewall.

3. The apparatus of claim 1 , wherein each cuvette holder includes a mounting feature configured to interface with an alignment feature of the plurality of alignment features to restrain a movement of the cuvette holder.

4. The apparatus of claim 1 , wherein each cuvette holder is configured to hold exactly eight cuvettes.

5. The apparatus of claim 1 , wherein the plate is configured to accommodate twelve cuvette holders on the first surface.

6. The apparatus of claim 1 , wherein each cuvette positioning feature is configured to restrain a movement of a cuvette that is inserted into the internal volume such that the portion of each cuvette that is inserted into the internal volume is in direct thermal contact with the sidewall.

7. The apparatus of claim 1 , wherein a surface of the fluid flowpath is defined, at least in part, by the plate.

8. The apparatus of claim 7 , wherein a surface of the fluid flowpath is further defined, at least in part, by a cover over a second surface of the plate.

9. The apparatus of claim 1 , wherein the fluid flowpath follows a serpentine path.

10. The apparatus of claim 1 , wherein the fluid flowpath is configured to cause at least a section of the first surface on which the cuvette holders are disposed to have a substantially uniform temperature when a heat transfer fluid flows through the fluid flowpath.

11. The apparatus of claim 10 , wherein a heat transfer fluid flows through the fluid flowpath and causes at least the section of the first surface to have a substantially uniform temperature.

12. The apparatus of claim 1 , further comprising a pump, wherein the pump is configured to flow the heat transfer fluid through the fluid flowpath.

13. The apparatus of claim 1 , wherein:

the cuvette holder is configured to hold N cuvettes and the plurality of second electrodes includes N second electrodes, and

each cuvette positioning feature is further configured to restrain a movement of a cuvette that is inserted into the internal volume such that:

a third portion of each cuvette that is inserted into the internal volume is electrically coupled to the first electrode, and

a fourth portion of each cuvette that is inserted into the internal volume is electrically coupled to one corresponding Nth second electrode.

14. The apparatus of claim 13 , wherein the sidewall is electrically conductive and is the first electrode.

15. The apparatus of claim 13 , wherein:

the third portion of each cuvette inserted into the internal volume is the portion of that cuvette, and

the fourth portion of each cuvette inserted into the internal volume is in thermal connection with the second sidewall.

16. The apparatus of claim 1 , further comprising switching circuitry, wherein:

each second electrode is electrically coupled with the switching circuitry, and

the switching circuitry is configured to electrically couple one second electrode to a power supply while the other second electrodes are not electrically coupled to the power supply.

17. The apparatus of claim 16 , further comprising a controller configured to control the switching circuitry, wherein the controller includes control logic for:

causing the switching circuitry to alternatively electrically couple each second electrode to the power supply while the other second electrodes are not electrically coupled to the power supply.

18. The apparatus of claim 17 , wherein the controller further includes control logic for:

measuring the resistance of a solution in a cuvette that has been inserted into the internal volume.

19. The apparatus of claim 18 , wherein the controller further includes control logic for:

adjusting, based on the measurement of the resistance of the solution in the cuvette that has been inserted into the internal volume, one or more of the following: the solution in that cuvette, the current to be applied to that cuvette, and the duration the current is to be applied to that cuvette.

20. The apparatus of claim 1 , further comprising a plurality of cuvettes, wherein:

each cuvette is inserted in the internal volume of the cuvette holder,

a movement of each cuvette is restrained by a corresponding cuvette positioning feature, and

each cuvette is electrically coupled to the first electrode and to one of the plurality of second electrodes.

21. The apparatus of claim 1 , further comprising a plurality of cuvettes, wherein:

each cuvette is inserted in the internal volume of the cuvette holder,

a movement of each cuvette is restrained by a corresponding cuvette positioning feature, and

a portion of each cuvette is in thermal connection with the sidewall.

22. The apparatus of claim 1 , wherein the apparatus is configured to place 96 cuvettes in thermal connection with the first surface of the plate at one time.

23. An apparatus comprising:

a plurality of cuvette holders, wherein each cuvette holder is configured to hold a plurality of cuvettes and includes a bottom and a sidewall that are in thermal connection with each other and at least partially define an internal volume of the cuvette holder, a first electrode, and a plurality of second electrodes that are electrically isolated from each other and from the first electrode;

a plate including a first surface; and

a fluid flowpath in thermal connection with the first surface, wherein:

the first electrode and the plurality of second electrodes are configured such that insertion of a cuvette into the internal volume causes a portion the cuvette to be in thermal connection with the sidewall and causes the cuvette to be electrically coupled to the first electrode and to one of the plurality of second electrodes, and

the bottom of each cuvette holder that is placed on the first surface is in thermal connection with the first surface.

24. The apparatus of claim 23 , wherein:

the cuvette holder is further configured to hold N cuvettes

the cuvette holder further includes N second electrodes, and

the first electrode and the plurality of second electrodes are further configured such that insertion of each cuvette into the internal volume causes each cuvette to be electrically coupled to one corresponding Nth second electrode.

Assignments (4)
RELEASE OF SECURITY INTEREST Recorded Jul 1, 2022
From: PERCEPTIVE CREDIT HOLDINGS II, LP, AS ADMINISTRATIVE AGENT
To: ZYMERGEN INC.
Reel/Frame 060421/0533 →
PATENT SECURITY AGREEMENT Recorded Dec 26, 2019
From: ZYMERGEN INC.
To: PERCEPTIVE CREDIT HOLDINGS II, LP, AS ADMINISTRATIVE AGENT
Reel/Frame 051425/0485 →
CORRECTIVE ASSIGNMENT TO CORRECT THE TYPOGRAPHICAL ERROR IN THE ASSIGNEE'S NAME, "ZYMERGEN INC.", PREVIOUSLY RECORDED ON REEL 046192 FRAME 0078. ASSIGNOR(S) HEREBY CONFIRMS THE THE CORRECT SPELLING OF THE ASSIGNEE'S NAME IS "ZYMERGEN INC." AS INDICATED ON THE ORIGINAL EXECUTED ASSIGNMENT. Recorded Feb 5, 2019
From: CESAREK, JOHN; LEE, TRAVIS; HANSEN, RICHARD
To: ZYMERGEN INC.
Reel/Frame 048261/0975 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jun 25, 2018
From: CESAREK, JOHN; LEE, TRAVIS; HANSEN, RICHARD
To: ZYMERGEN, INC.
Reel/Frame 046192/0078 →
Continuity (1)
Related Publication 20180002652A1 · Jan 4, 2018