Apparatus for Generating Electrical Pulses and Methods of Using Same

A pulse generator for generating electrical pulses comprising:

• a first circuit for generating a first pulse having a long duration and low voltage amplitude;
• a second circuit for generating a second pulse having a short duration and high voltage amplitude;
• and
• a control circuit for controlling the timing of said first circuit and said second circuit to respectively generate said first pulse and said second pulse.

A pulse generator for generating electrical pulses comprising:

• first generator means for generating a first pulse having a long duration and low voltage amplitude;
• second generator means for generating a second pulse having a short duration and high voltage amplitude;
• and
• control means for controlling timing of the pulses generated by said first generator means and said second generator means.

A method of enhancing gene expression using a pulse generator, the method comprising the sequential, the non-sequential, and the sequence independent steps of:

• triggering a first pulse having a long duration and low voltage amplitude from a first circuit of the pulse generator;
• delivering the first pulse to at least one cell to cause electroporation at the plasma membrane of the at least one cell;
• triggering a second pulse having a long duration and low voltage amplitude from a second circuit of the pulse generator;
• and
• delivering the second pulse to the at least one cell to cause electroporation at the nuclear membrane of the at least one cell.

A method of enhancing gene expression in a cell using a multi-pulse generator, the method comprising the sequential, the non-sequential, and the sequence independent steps of:

• charging a capacitor;
• triggering a high voltage, high current transistor to initiate discharge of the charge accumulated in the capacitor into at least one cell to cause electroporation at the plasma membrane of the at least one cell;
• triggering the high voltage, high current transistor to stop the discharge of the capacitor after a predetermined long duration;
• actuating a switch to decouple the capacitor from the at least one cell;
• charging a transmission line;
• triggering a high voltage switch to initiate discharge of the charge accumulated in the transmission line into the at lest one cell to cause electroporation at the nuclear membrane of the at least one cell;
• and
• triggering the high voltage switch to stop discharge of the transmission line after a predetermined short duration.

A multi-pulse generator for causing electroporation at both a cellular plasma membrane and a nuclear membrane, said multi-pulse generator comprising:

• means for triggering a first pulse having a long duration and low voltage amplitude;
• means for delivering said first pulse to at least one cell, said first pulse causing electroporation at the plasma membrane of the at least one cell;
• means for triggering a second pulse having a short duration and high voltage amplitude;
• and
• means for delivering the second pulse to the at least one cell, said second pulse causing electroporation at the nuclear membrane of the at least one cell.

A multi-pulse generator for enhancing gene expression in a cell, said multi-pulse generator comprising:

• means for accumulating a charge;
• means for selectively discharging the accumulated charge into at least one cell to cause electroporation at the plasma membrane of the at least one cell, and terminating discharge of the accumulated charge;
• means for charging a transmission line;
• and
• means for selectively discharging said transmission line into the at least one cell to cause electroporation at the nuclear membrane of the at least one cell, and terminating discharge of the transmission line.

A dual-pulse generator for enhancing gene expression in a cell, said dual-pulse generator comprising:

• a first pulse generator for generating a first pulse having a long duration and low voltage amplitude, said first pulse causing electroporation of the cellular plasma membrane of the cell;
• a second pulse generator for generating a second pulse having a short duration and high voltage amplitude, said second pulse causing electroporation of the nuclear membrane of the cell;
• and
• a control circuit for controlling timing of pulses generated by said first pulse generator and said second pulse generator.

A dual-pulse generator for enhancing gene expression in a cell, the dual-pulse generator comprising:

• first generator means for generating a first pulse having a long duration and low voltage amplitude, said first pulse causing electroporation of the cellular plasma membrane of the cell;
• second generator means for generating a second pulse having a short duration and high voltage amplitude, said second pulse causing electroporation of a nuclear membrane of the cell;
• and
• control means for controlling timing of the pulses generated by said first generator means and said second generator means.

A dual-pulse generator for enhancing gene expression in a cell, the dual-pulse generator comprising:

• a first high voltage power supply;
• a first charging resistor coupled to said first high voltage power supply;
• a capacitor coupled at a first end to said first charging resistor and coupled at a second end to a load;
• a transistor coupled to the second end of the first charging resistor and the first end of the capacitor, said transistor controlling electrical discharge of the capacitor to said load;
• a second high voltage power supply;
• a second charging resistor coupled to said second high voltage power supply;
• a transmission line coupled at a first end to said second charging resistor and coupled at a second end to the load;
• a control circuit for controlling electrical discharge of said capacitor and said transmission line;
• a first switch for selectively coupling and decoupling said capacitor from the load;
• a first trigger unit coupled to said control circuit and said first switch, said first trigger unit actuating said first switch responsive to one or more commands received from said control circuit;
• a second switch for selectively discharging said transmission line;
• a second trigger unit coupled to said control circuit and said second switch, said second trigger unit actuating said first switch responsive to one or more commands received from said control circuit;
• and
• a delivery apparatus for delivering the electrical discharges from said capacitor and said transmission line to the load.

A method of enhancing gene expression using a dual-pulse generator, the method comprising the sequential, the non-sequential, and the sequence independent steps of:

• triggering a first pulse type from the dual-pulse generator, the first pulse type having a long duration and low voltage amplitude;
• delivering the first pulse type to at least one cell to cause causes electroporation at the plasma membrane of the at least one cell;
• waiting a predetermined time interval;
• triggering a second pulse type from the dual-pulse generator, the second pulse type having a short duration and high voltage amplitude;
• and
• delivering the second pulse type to the at least one cell to cause electroporation at the nuclear membrane of the at least one cell.