US 5,869,326 A · Electroporation employing user-configured pulsing scheme

Abstract

An electroporation method and apparatus generating and applying an electric field according to a user-specified pulsing scheme. Advantageously, one such pulse includes a low voltage pulse of a first duration, immediately followed by a high voltage of a second duration, immediately followed by a low voltage of a third duration. The low voltage electroporation field accumulates molecules at the surface of a cell, the appropriately high voltage field creates an opening in the cell, and the final low voltage field moves the molecule into the cell. The molecules may be DNA, portions of DNA, chemical agents, the receiving cells may be eggs, platelets, human cells, red blood cells, mammalian cells, plant protoplasts, plant pollen, liposomes, bacteria, fungi, yeast, sperm, or other suitable cells. The molecules are placed in close proximity to the cells, either in the interstitial space in tissue surrounding the cells or in a fluid medium containing the cells.

Claims21

Dependent claims

An apparatus for generating an electroporation pulse shape, said apparatus comprising:

a first power supply to provide a first output voltage;
a second power supply to provide a second output voltage, wherein the first and second power supplies are separate sources of electrical energy;
a transformer having electromagnetically coupled primary and secondary windings, said secondary winding being interposed between a pair of output terminals;
a first switch coupled to the first power supply and the primary winding and being responsive to a first gating signal to apply the first output voltage from the first power supply to the primary winding;
a second switch coupled to the second power supply and the secondary winding and responsive to a second gating signal to apply the second output voltage from the second power supply to the secondary winding;
and
a controller to receive user specification of an output pulse shape and provide the first and second gating signals to generate the specified output pulse shape at the output terminals.

4 more elements

An electroporation pulse generating apparatus, comprising:

a transformer including a primary winding and a secondary winding;
a first electrical energy source coupled to the primary winding to place a first predetermined voltage across primary winding in response to a first gating signal;
a second electrical energy source coupled to the secondary winding to place a second predetermined voltage across second winding in response to a second gating signal wherein the first and second electrical energy sources are separate sources of electrical energy;
a controller, electrically coupled to the first and second energy sources, and programmed to perform method steps comprising:
(a) to generate a first voltage at the secondary winding terminals by applying a first voltage to the secondary winding terminals of the transformer;
(b) a predetermined delay after initiating step (a), to maintain application of the first voltage at the secondary winding terminals while concurrently applying a second voltage to the primary winding terminals;
and
(c) a predetermined delay after initiating step (b), to cease application of the second voltage to the primary winding terminals while maintaining application of the first voltage at the secondary winding terminals for a predetermined duration.

5 more elements

Dependencies are inferred from the claim text.

Electroporation employing user-configured pulsing scheme

Life of this patent

Electroporation employing user-configured pulsing scheme

Life of this patent

Abstract

Claims21