Microfluidic vortex-assisted electroporation system and method

Patent metadata

Field	Value
Publication number	US-9029109-B2
Application number	US-201313961084-A
Country	US
Kind code	B2
Filing date	Aug 7, 2013
Priority date	Aug 7, 2013
Publication date	May 12, 2015
Grant date	May 12, 2015

Abstract

A system and method include delivering cells of interest to multiple traps via a channel connecting the traps, maintaining a vortex flow in the traps to trap the cells of interest in the traps, providing first molecules of interest to the traps, and providing an electric field across the traps to perform electroporation of the first molecules of interest into the cells of interest in the traps.

First claim

The invention claimed is:

1. A method comprising:
delivering cells of interest to multiple traps via fluid flowing along an axis through a channel connecting the traps;
maintaining a vortex flow in the traps to trap the cells of interest in the traps, wherein the vortex comprises a rotational flow around an axis perpendicular to axis of flow through the channel;
providing first molecules of interest to the traps; and
providing an electric field across the traps to perform electroporation of the first molecules of interest into the cells of interest in the traps.

2. The method of claim 1 wherein delivering cells of interest is performed by transporting a first fluid solution containing the cells of interest via the channel at a speed such that the fluid has a Reynolds number of greater than 100 to create the vortex flow in the traps.

3. The method of claim 2 wherein providing first molecules of interest to the traps comprises using a second fluid solution containing the molecules of interest while maintaining the vortex flow in the traps and removing the first solution.

4. The method of claim 3 wherein the electric field across the traps is substantially uniform.

5. The method of claim 3 and further comprising using a third fluid solution containing further molecules of interest while maintaining the vortex flow in the traps, the further molecules of interest being provided following electroporation of the first molecules of interest.

6. The method of claim 5 and further comprising providing an electric field across the traps to perform electroporation of the second molecules of interest into the cells of interest in the traps, wherein the electric field is adapted to enhance delivery of the second molecules of interest to the cells.

7. The method of claim 6 and further comprising replacing the second fluid with additional fluids containing additional molecules of interest while maintaining the vortex flow in the traps.

8. The method of claim 6 wherein the vortex flow is maintained during the entire method.

9. The method of claim 2 wherein delivering cells of interest in solution to multiple traps via the channel connecting the traps includes providing the first solution to the channel via an inertial focusing region to cause the cells of interest to move close to the sides of the channel via fluidic forces.

10. The method of claim 9 wherein the channel breaks into multiple channels, each having opposing pairs of traps disposed along a length of the channels.

11. A system comprising:
a first channel having an inertial focusing region to move cells in a solution travelling through the first channel towards sides of the channel;
a plurality of serial opposed pairs of traps disposed along a length of the first channel downstream of the inertial focusing region, the size of each trap adapted to promote vortex flow within the traps while the solution is flowing through the first channel to trap the cells in the traps;
an outlet of the channel disposed downstream from the plurality of serial opposed pairs of traps; and
electrodes coupled to ends of the opposed traps to apply an electric field across the traps suitable for electroporation of molecules into the cells.

12. The system of claim 11 wherein the channel has a width such that cells traveling through the channel are approximately 30 percent or greater than the width of the channel.

13. The system of claim 11 wherein the inertial focusing region of the channel is approximately 0.7 cm or longer.

14. The system of claim 11 wherein a second channel configured the same as the first channel is in parallel with the first channel.

15. The system of claim 14 and further comprising multiple inlets to the first and second channel inertial focusing regions, the inlets adapted to receive solutions from multiple sources including a cell solution and a molecule solution.

16. The system of claim 15 wherein the inlets are adapted to receive further solutions for incubation and flushing.

17. The system of claim 14 wherein the first and second channels each include five opposing pairs of traps serially disposed downstream from the inertial focusing region of each channel.

18. The system of claim 14 wherein each trap is approximately square in shape with each opposed pair of traps forming a rectangle having a channel entrance and exit bisecting the opposed pair of traps.

19. The system of claim 18 wherein the traps have sides of between approximately 1 mm to 500 μm.

20. The system of claim 14 wherein the electrodes comprise multiple electrode tips adapted to create a substantially uniform electric field, and wherein traps from the first channel adjacent traps from the second channel share an electrode of one polarity.

21. A system comprising:
a first channel having an inertial focusing region to move cells in a solution travelling through the first channel towards sides of the channel;
a plurality of traps disposed along a length of the first channel downstream of the inertial focusing region, the size of each trap adapted to promote vortex flow within the traps while the solution is flowing through the first channel to trap the cells in the traps;
an outlet of the channel disposed downstream from the plurality of traps; and
electrodes coupled to ends of the traps to apply an electric field across the traps suitable for electroporation of molecules into the cells.

Assignees

• Harvard College

Inventors

• Hur Soojung Claire
• Yun Hoyoung

Classifications

• C12N13/00 Primary

  Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves · CPC title

• C12M35/02 Primary

  Electrical or electromagnetic means, e.g. for electroporation or for cell fusion · CPC title

• C12N15/87

  Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation · CPC title

• B01L3/502715

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

• B01L3/502761

  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