Methods and devices related to a regulated flow electroporation chamber

• US 9,546,350 B2
• Filed: 08/10/2010
• Issued: 01/17/2017
• Est. Priority Date: 05/12/2004
• Status: Active Grant

• Current Assignee: Maxcyte, Inc. (Entremed, Incorporated)
• Original Assignee: Maxcyte, Inc. (Entremed, Incorporated)
• Inventors: Dzekunov, Sergey, Chopas, Nicholas, Li, Linhong
• Primary Examiner(s): Beisner, William H
• Application Number:
  US12/853,772
• Publication Number:
  US 20110065171A1
• Time in Patent Office: 2,352 Days
• Field of Search: 435/173.6, 435/285.2
• US Class Current: 1/1
• CPC Class Codes: B01L 3/502 with fluid transport, e.g. in multi-compartment structures centrifugal-type cuvettes G01N21/07; analysis by separation into components G01N30/00; automatic analysers G01N35/00
• CPC Class Codes: B01L 3/5027 by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: C12M 35/02 Electrical or electromagnetic means, e.g. for electroporation or for cell fusion
• CPC Class Codes: C12M 41/00 Means for regulation, monitoring, measurement or control, e.g. flow regulation controlling or regulating chemical, physical or physicochemical processes B01J19/0006; heating or cooling apparatus for laboratory use B01L7/00; electro optical investigation of individual particles, flow cytometers G01N15/14; automatic analysis G01N35/00; controlling or regulating in general G06N

Abstract

The electroporation chamber and its related devices combine the features of an electroporation chamber that acts as a manifold for regulation of sample flow with those of a flow electroporation device to form a regulated flow electroporation device. The invention further comprises a novel regulated flow electroporation chamber that enables conditions in which a sample is uniformly processed in individual fractions or volumes in a fully closed (sterile) system.

Claims

1. A flow electroporation chamber comprising:

(a) at least two electrodes separated by a spacer, the electrodes and the spacer configured to form a single, continuous chamber within the spacer; and

(b) at least three ports configured to traverse the spacer to communicate through the chamber within the spacer, wherein

(i) a first port is for sample flow into the chamber;

(ii) a second port is for processed sample flow out of the chamber; and

(iii) a third port is for non-sample fluid or gas flow in or out of the chamber,

wherein the non-sample fluid or gas flows out of the chamber when a sample flows into the chamber and the non-sample fluid or gas flows into the chamber when processed sample flows out of the chamber.

2. The chamber of claim 1, wherein a first port is in fluid communication with one or more sample container.

3. The chamber of claim 1, wherein a second port is in fluid communication with one or more processed sample container.

4. The chamber of claim 1, wherein a third port is in fluid communication with a reservoir container, wherein the reservoir container contains all or part of the non-sample fluid or gas volume when the chamber is filled or partially filled with a sample volume.

5. The chamber of claim 1, having at least four ports.

6. The chamber of claim 5, wherein at least a fourth port is in fluid communication with the sample container, the processed sample container, the non-sample gas or fluid reservoir or a reagent container.

7. A flow electroporation device comprising:

a) at least one sample container in fluid communication with an electroporation chamber of claim 1 through a first chamber port, forming a first fluid path; and

b) at least one processed sample container in fluid communication with the electroporation chamber of claim 1 through a second chamber port, forming a second fluid path.

8. The device of claim 7, further comprising at least one non-sample fluid or gas reservoir in fluid communication with the electroporation chamber through a third chamber port, forming a third fluid path.

9. The device of claim 8, wherein at least one fluid path is a closed fluid path.

10. The device of claim 9, wherein the first, second and third fluid paths are closed fluid paths.

11. The device of claim 8, wherein the second fluid path is in fluid communication with the sample container of the first fluid path and the non-sample reservoir of the third fluid path distal to the position of the electroporation chamber in relation to the processed sample container in the second fluid path.

12. The device of claim 8, further comprising at least one pump operatively coupled to at least one fluid path.

13. The device of claim 12, wherein the pump is a peristaltic pump.

14. The device of claim 13, wherein the peristaltic pump is a full compression peristaltic pump.

15. The device of claim 8, wherein the sample container is collapsible, expandable or fixed volume.

16. The device of claim 8, wherein the processed sample container is collapsible, expandable or fixed volume.

17. The device of claim 8, wherein fluid flow is modulated by further comprising one or more valves.

18. The device of claim 7, wherein the at least one sample container, the at least one processed sample container, or both the at least one sample container and the at least one processed sample container further comprises at least one non-sample gas or fluid reservoir.