Systems, apparatus, and methods for droplet-based microfluidics cell poration

Patent metadata

Field	Value
Publication number	US-2016108433-A1
Application number	US-201514979705-A
Country	US
Kind code	A1
Filing date	Dec 28, 2015
Priority date	Jun 27, 2013
Publication date	Apr 21, 2016
Grant date	—

Abstract

The present disclosure provides systems, apparatus, and methods that include an electrowetting-on-dielectric microfluidic apparatus for effecting poration of cells in a droplet.

First claim

We claim:

1 . An electrowetting-on-dielectric microfluidic apparatus for cell poration comprising:
a first plate; a second plate spaced-apart from the first plate and defining a flow channel therebetween for receiving a droplet having cells therein; an acoustic energy generator in communication with the first plate for providing sonoporation to cells within a droplet received in the flow channel; and a resilient member in communication with the acoustic energy generator and in communication with a droplet received in the flow channel to provide acoustic streaming and sonoporation to cells within a droplet.

2 . The apparatus according to claim 1 , wherein the resilient member is a deformable membrane carried by the first plate and in communication with a droplet received in the flow channel.

3 . The apparatus according to claim 1 , wherein the resilient member is a bubble partially enclosed by a recess defined by the first plate, wherein the bubble is in communication with a droplet received in the flow channel.

4 . The apparatus according to claim 1 , wherein the first plate comprises a hydrophobic layer on a metal film, and wherein the second plate comprises a hydrophobic layer on a dielectric film on a patterned metal electrode.

5 . The apparatus according to claim 1 , wherein the acoustic energy generator is a piezoelectric disk translator.

6 . The apparatus according to claim 1 , further including an electric field generator in electrical field communication with the flow channel for providing electroporation to cells within the droplet received in the flow channel.

7 . The apparatus according to claim 6 , wherein the electric field generator comprises electrodes placed between the first plate and the second plate, and wherein the electrodes are spaced-apart a distance generally corresponding to a width of the resilient member.

8 . The apparatus according to claim 6 , wherein the acoustic energy generator and the resilient member comprise an immersion transducer extending from the first plate into the flow channel to provide acoustic streaming and sonoporation to cells within a stationary droplet received in the flow channel.

9 . The apparatus according to claim 1 , wherein a droplet received in the flow channel is a stationary droplet.

10 . A system comprising:
an electrowetting-on-dielectric microfluidic apparatus for cell poration that includes:
a first plate;
a second plate spaced-apart from the first plate and defining a flow channel therebetween for receiving a droplet having cells therein;
an acoustic energy generator in communication with the first plate; and
a resilient member in communication with the acoustic energy generator and in communication with a droplet received in the flow channel; and
a control module configured for:
controlling the acoustic energy generator to apply energy to the apparatus for providing acoustic streaming and sonoporation to cells within a droplet.

11 . The system according to claim 10 , wherein the resilient member is a deformable membrane that is in communication with a droplet received in the flow channel.

12 . The system according to claim 10 , wherein the resilient member is a bubble partially enclosed by a recess defined by the first plate, wherein the bubble is in communication with a droplet received in the flow channel.

13 . The system according to claim 10 , wherein the acoustic energy generator and the resilient member comprise an immersion transducer extending from the first plate into the flow channel to provide acoustic streaming and sonoporation to cells within a stationary droplet received in the flow channel.

14 . The system according to claim 10 , wherein the first plate comprises a hydrophobic layer on a metal film, and wherein the second plate comprises a hydrophobic layer on a dielectric film on a patterned metal electrode.

15 . The system according to claim 10 , wherein the acoustic energy generator is a piezoelectric disk translator.

16 . The system according to claim 10 , wherein a droplet received in the flow channel is a stationary droplet.

17 . The system according to claim 10 , wherein the apparatus further includes an electric field generator in electrical field communication with the flow channel, and wherein the control module is further configured for controlling the electric field generator to apply an electric field for providing electroporation to cells within the droplet.

18 . The system according to claim 17 , wherein the electric field generator comprises electrodes placed between the first plate and the second plate, and wherein the electrodes are spaced-apart a distance generally corresponding to a width of the resilient member.

19 . The system according to claim 18 , wherein the control module is configured for:
controlling the acoustic energy generator to apply energy to the apparatus for providing acoustic streaming and sonoporation to cells within a droplet over a first time period; and controlling the electric field generator to apply an electric field for providing electroporation to cells within a droplet over a second time period.

20 . The system according to claim 19 , wherein the first time period and the second time period are simultaneous.

21 . The system according to claim 19 , wherein the first time period and the second time period partially overlap.

22 . The system according to claim 19 , wherein the first time period and the second time period are in sequence.

23 . A method comprising:
in a spaced-apart plate arrangement defining a flow channel therebetween, applying an acoustic energy field to a resilient member in communication with a droplet having cells therein to induce an ultrasonic vibration in the resilient member for providing acoustic streaming and sonoporation of the cells.

24 . The method according to claim 23 , wherein the resilient member is a deformable membrane.

25 . The method according to claim 23 , wherein the resilient member is a bubble.

26 . The method according to claim 23 , further comprising applying an electric field to the droplet having cells therein to effect electroporation of the cells.

27 . The method according to claim 26 , wherein the electric field is applied generally perpendicularly to the acoustic energy field and the acoustic streaming focuses the cells within the electric field to provide an electric field thereto.

28 . The method according to claim 23 , wherein a droplet received in the flow channel is a stationary droplet.

Assignees

• Univ Duke

Inventors

• Fair Richard
• Madison Andrew
• Royal Matthew

Classifications

• C12M23/16 Primary

  Microfluidic devices; Capillary tubes (integrated microfluidic structures B01L3/5027; microreactors B01J19/0093) · CPC title

• B01L3/502715

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

• C12N15/87 Primary

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

• B01L2400/0427

  Electrowetting · CPC title

• B01L2300/165

  Specific details about hydrophobic, oleophobic surfaces · CPC title