We are first to demonstrate experimentally that endocytotic vesicles are electropermeabilized by nanosecond, high-voltage electric pulses. The method could be used to introduce various substances into living cells. In contrast to some previous publications in the field of nanosecond electropermeabilization, we have shown that the permeabilization of internal cell membranes at given electrical parameters is accompanied by a permeabilization of plasma membrane.
COBISS.SI-ID: 7642708
We are first to demonstrate experimentally that DNA/membrane interaction and gene expression are affected by reversing the polarity and by increasing the repetition rate of electric pulses in gene electrotransfer. The obtained results reveal the existence of metastable and stable DNA/membrane complex. We showed that only DNA belonging to the stable complex leads to effective gene expression and that the formation time of DNA/membrane complex is of the order of a second, which can be used to improve protocols of electromediated gene delivery.
COBISS.SI-ID: 7524436
We developed and tested a Blumlein generator for highvoltage (up to 1000 V) nanosecond pulses with high repetition rate (up to 100 kHz), variable duration (from 30 to 200 ns) and polarity. We developed a microelectrodes that allow us to generate welldefined and relatively homogeneous electric field in a 100 micrometer gap. Both devices together enable us to apply nanosecond pulses and to observe its affects on cells under the microscope. With the developed generator we showed that we can electropermeabilize endocytotic vesicles and inevitably also the plasma membrane.
COBISS.SI-ID: 7346772
We determined experimentally the relations between the amplitude and duration/number of pulses resulting in the same fraction of electroporated cells. Pulse duration was varied from 150 ns to 100 ms, and the number of pulses was varied from 1 to 128. We are first to show that the model of electroporation, based on physical quantities, can describe the measured data on the interval of pulse durations from nano- to miliseconds. Such relations between pulse parameters can be important in planning of electroporation-based treatments, such as electrochemotherapy and nonthermal irreversible electroporation, when recommended values of pulse parameters cannot be set.
COBISS.SI-ID: 8714836
We developed prototype of tip electrode chamber for electropermeabilization. Tip electrode chamber is used for quick and efficient permeabilization of small and controlled volumes of cell suspensions. Tip electrode chamber allow delivery of electric field in different directions thus increasing the membrane area of cell being permeabilized and therefore efficiency of electropermeabilization. We tested the tip electrode chamber for gene transfection and concluded that by changing the electric field direction in tip electrode chamber we can increase the fraction of transfected cells.
COBISS.SI-ID: 8590164