# US 8,465,955 B2 · Method for electroporation of biological cells

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- Number: US 8,465,955 B2
- Document ID: US8465955B2
- Status: Granted
- Kind: B2
- Application number: 12/072,016
- Filed: 2008-02-22
- Published: 2008-09-11
- Granted: 2013-06-18
- Earliest claimed: 2002-01-21
- Assignees: Eppendorf AG
- Applicants: Ulrich Zimmermann; Vladimir Sukhorukov; Kurt Lucas
- Inventors: Ulrich Zimmermann; Vladimir Sukhorukov; Kurt Lucas
- Web page: https://patentgrep.com/patent/US8465955B2

## CPC

- C12M 35/02: {Electrical or electromagnetic means, e.g. for electroporation or for cell fusion}
- C12N 13/00: Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
- C12M: APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; {APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS}
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

Described are methods for electrical treatment of biological cells, in particular using electrical field pulses, involving the steps: arrangement of the cells (1) on apertures (2) of a solid planar carrier element (3) which divides a measuring chamber (10) into two compartments (11, 12); and temporary formation of an electrical treatment field which permeates the cells, wherein an alternating-current impedance measurement takes place on the carrier element (3), and from the result of the alternating-current impedance measurement, a degree of coverage of the carrier element and/or healing of the cells after electrical treatment are/is acquired. Also described are devices for implementing the methods.

## Claims

### Claim 1

A method for electrical treatment of biological cells using electrical field pulses comprising the steps of:
arranging the cells on apertures of a solid planar carrier element which divides a measuring chamber into two compartments, said carrier element being covered with said cells with a degree of coverage;
temporarily forming an electrical treatment field which permeates the cells,
measuring an alternating-current impedance on the carrier element, wherein the measuring step is at a predetermined measuring frequency at which frequency the alternating-current impedance measured on the carrier element is increased to its maximum as a result of the arrangement of the cells, and
determining said degree of coverage of the carrier element or a degree of healing of said cells after said electrical treatment, or both from the result of said alternating-current impedance measurement.

### Claim 2 (depends on 1)

The method according to claim 1, wherein the measuring step includes measuring an alternating current impedance kinetic and measuring at least one impedance value.

### Claim 3 (depends on 2)

The method according to claim 2, wherein said degree of coverage of the carrier element is determined by said measuring the alternating-current impedance kinetic during arrangement of the cells on the apertures of the carrier element, or by measuring the at least one impedance value, and calculating the degree of coverage from at least one comparison value.

### Claim 4 (depends on 2)

The method according to claim 2 wherein said degree of healing is determined by said measuring the alternating-current impedance kinetic after electrical treatment of the cells or by measuring the at least one impedance value at at least one predetermined point in time, and comparing with at least one reference value.

### Claim 5 (depends on 1)

The method according to claim 1, wherein said measuring frequency is determined by a preliminary test in which a frequency dependence of the alternating-current impedance is determined from at least one reference value.

### Claim 6 (depends on 1)

The method according to claim 1 further comprising observing optically, with a microscope, the arrangement of the cells and the result of the electrical treatment.

### Claim 7 (depends on 1)

The method according to claim 1 wherein the arranging step comprises adsorbing said cells from a suspension or an adherent growth of said cells.

### Claim 8 (depends on 1)

The method according to claim 1 further comprising cultivating said cells on the carrier element after the forming step.