Electrosonic cell manipulation device and method of use thereof

• US 7,704,743 B2
• Filed: 03/28/2006
• Issued: 04/27/2010
• 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: Degertekin, Fahrettin L., Fedorov, Andrei G.
• Primary Examiner(s): Ketter; James S
• Application Number:
  US11/277,662
• Publication Number:
  US 20060223185A1
• Time in Patent Office: 1,491 Days
• Field of Search: None
• US Class Current: 435/461
• 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. A method of injecting a substance into a living cell having a cell membrane, comprising the steps of:

a. placing the substance, the cell and a liquid into a tapering passage; and

b. applying an energy to the cell sufficient to induce poration of the cell, wherein the step of applying an energy to the cell, comprises applying an acoustic pressure wave to the liquid, thereby inducing sonoporation in the cell wherein the tapering passage includes a narrow end that defines an opening passing therethrough, wherein the method further comprises the step of forcing the cell through the opening so that as the cell passes through the opening the cell membrane allows the substance to pass therethrough.

2. The method of claim 1, wherein the step of applying an acoustic pressure wave further comprises the step of forming a standing wave within the tapering passage.

3. The method of claim 1, wherein the acoustic wave comprises an ultrasonic wave.

4. The method of claim 1, wherein the step of applying an energy to the cell comprises the step of focusing the energy within the tapering passage, thereby applying a highly localized energy gradient to the cell.

5. The method of claim 1, wherein the substance comprises genetic material.

6. The method of claim 1, wherein the substance comprises a predetermined chemical composition.

7. The method of claim 1, further comprising the step of applying an electric field to the cell, thereby inducing electroporation in the cell.

8. The method of claim 7, wherein the step of applying an electric field to the cells comprises applying an oscillating electric field.

9. A method of injecting a substance into a living cell having a cell membrane, comprising the steps of:

a. placing the substance, the cell and a liquid into a tapering passage; and

b. applying an oscillating electric field to the cell, thereby inducing electroporation in the cell to the cell sufficient to induce poration of the cell; and

c. applying an acoustic pressure wave to the liquid, thereby inducing sonoporation in the cell.

10. The method of claim 9, wherein the step of applying an acoustic pressure wave further comprises the step of forming a standing wave within the tapering passage.

11. The method of claim 9, wherein the acoustic wave comprises an ultrasonic wave.

12. The method of claim 9, wherein the step of applying an energy to the cell comprises the step of focusing the energy within the tapering passage, thereby applying a highly localized energy gradient to the cell.

13. The method of claim 9, wherein the substance comprises genetic material.

14. The method of claim 9, wherein the substance comprises a predetermined chemical composition.