Electrosonic cell manipulation device

• US 8,334,133 B2
• Filed: 03/12/2010
• Issued: 12/18/2012
• Est. Priority Date: 03/30/2005
• Status: Active Grant

• Current Assignee: Georgia Tech Research Corporation (University System of Georgia)
• Original Assignee: Georgia Tech Research Corporation (University System of Georgia)
• Inventors: Fedorov, Andrei G., Degertekin, Fahrettin L.
• Primary Examiner(s): Ketter, Jim
• Application Number:
  US12/723,033
• Publication Number:
  US 20100227371A1
• Time in Patent Office: 1,012 Days
• Field of Search: None
• US Class Current: 435/285.2
• CPC Class Codes: C12M 35/02 Electrical or electromagnetic means, e.g. for electroporation or for cell fusion
• CPC Class Codes: C12M 35/04 Mechanical means, e.g. sonic waves, stretching forces, pressure or shear stimuli
• CPC Class Codes: C12M 47/04 Cell isolation or sorting purging biological preparations of unwanted cells C12N5/0081, determining the presence or kind of microorganism C12Q1/04
• CPC Class Codes: C12M 47/06 Hydrolysis; Cell lysis; Extraction of intracellular or cell wall material lysis of microorganisms C12N1/06; extracting or separating nucleic acids from biological samples C12N15/1003
• CPC Class Codes: C12N 13/00 Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves

Abstract

In method of injecting a substance into a living cell having a cell membrane, the substance, the cell and a liquid are placed into a tapering passage. Energy is applied to the cell, thereby inducing poration. To sort cells, a cellular suspension is placed in a tapering passage, including a narrow end that defines an opening that has a dimension corresponding to a cell size. An acoustic wave is applied, thereby forcing cells having a cell size smaller than the selected cell size through the opening, with a portion of the cells having a cell size not smaller than the selected cell size not forced through the opening. To extract material from a cell, an electric field and an acoustic wave are applied, thereby causing the cell membrane to allow the material to pass out of the cell.

Claims

1. An apparatus for manipulating cells, comprising:

a. a substrate, having a first side and an opposite second side, the substrate defining at least one tapering passage passing therethrough, the tapering passage opening to the first side with a wide end and also opening to the second side with a narrow end, the narrow end having a size corresponding to a size of a selected cell;

b. a first poration electrode and a spaced-apart second poration electrode disposed so as to impart a predetermined electrical field on the passage when an electrical potential difference is applied between the first poration electrode and the second poration electrode;

c. a fluid driving structure that drives fluid through the opening, the fluid driving structure including an ultrasonic transducer;

d. an oscillating circuit that applies an oscillating electric potential to the ultrasonic transducer, thereby causing the ultrasonic transducer to generate a acoustic wave in the tapering passage,

wherein the acoustic wave and the electrical field impart energy on at least a portion of the cells so as to cause at least one of cell poration, cell transfection, cell sorting or cell lysis.

2. The apparatus of claim 1, wherein the ultrasonic transducer is spaced apart from the wide end.

3. The apparatus of claim 1, wherein the ultrasonic transducer is disposed adjacent to the substrate.

4. The apparatus of claim 1, wherein the substrate comprises a silicon wafer.

5. The apparatus of claim 1, wherein the tapering passage comprises a pyramidal passage.

6. The apparatus of claim 1, wherein the tapering passage comprises a frusto-conical passage.

7. The apparatus of claim 1, wherein the first poration electrode and the second poration electrode each comprise a conductive substance disposed on an inside surface of the tapering passage.

8. The apparatus of claim 1, wherein the second poration electrode is disposed adjacent to the second side of the substrate.

9. The apparatus of claim 8, further comprising an insulator layer disposed adjacent to the second poration electrode and wherein the first poration electrode is disposed adjacent to the insulator layer.

10. The apparatus of claim 1, wherein the substrate includes a dopant so that the substrate comprises the second poration electrode.

11. The apparatus of claim 1, wherein the ultrasonic transducer comprises a capacitive transducer.

12. The apparatus of claim 11, wherein the capacitive transducer comprises a first transducer electrode and spaced-apart second transducer electrode so that when a potential is applied between the first transducer electrode and the second transducer electrode, the second transducer electrode moves relative to the first transducer electrode.

13. The apparatus of claim 1, wherein the ultrasonic transducer comprises a piezoelectric transducer.

14. The apparatus of claim 13, wherein the piezoelectric transducer comprises a layer of a piezoelectric material disposed between a first transducer electrode and an opposite second transducer electrode and oriented so that when an electric field is applied between the first transducer electrode and the second transducer electrode, a dimensional characteristic of the layer of piezoelectric material changes.

15. The apparatus of claim 1, wherein the substrate defines an array of tapering passages passing therethrough.

16. The apparatus of claim 15, wherein the array of tapering passages includes a first plurality of tapering passages and a spaced-apart second plurality of tapering passages, the apparatus further comprising a wall extending from the substrate so as to separate the first plurality of tapering passages from the second plurality of tapering passages.

17. The apparatus of claim 15, wherein the second poration electrode comprises a plurality of sub-electrodes, wherein each sub-electrode is disposed adjacent to a different set of the tapering passages and wherein each sub-electrode may be driven by a different electrical potential.