bioMarkeMagazine16.

qxp 14/11/2006 9:57 Page 14

Electroporation
Electroporation is the process of Electroporation as a method of gene transfer has a number of advantages over conventional
applying electrical field (pulses) to methods of cell permeablisation. It is a non-invasive, non-chemical method, and does not alter target
cell structure or function. This method is fast and easy to perform and unlike other chemical or
a living cell for a brief duration of biological methods it is relatively non-toxic. The electroporation technique can be applied to a much
time. This electrical field causes a wider selection of cell types because it is a physical method.
transient reversible breakdown of
the cellular membrane. This results
in the formation of pores which
allow exogenous molecules, such
as DNA, siRNA, proteins or drugs
to enter the cell. This process can
be used for transient and/or stable
transfections of mammalian cells.

Introduce molecules to Apply the pulse, pores After the pulse, the pores
the cells. form in the cells and reseal and the molecules
the molecules enter. remain in the cell.

This process involves several variables. The first two are field strength and pulse length which can be
controlled for efficient gene transfer. Field strength is simply the measured voltage applied across the
electrode gap and is expressed as kV/cm. In general, field strength is the voltage required to induce
membrane breakdown, which can be adjusted by varying the voltage applied or by changing the
distance of the gap between electrodes. The pulse length in general, is the duration of time the
sample is exposed to the electrical field and is measured in microsecond to milliseconds. The pulse
length is related to the size and how long the pores are open during the pulse and can be varied in
order to increase efficiency. The third variable is pulse shape, which falls into two forms, exponential
or square wave, which are generated by partial or complete discharge of a capacitor. The exponential
decay wave is used for both mammalian and bacterial electroporation; it provides high
transformation efficiencies but decreased viability of mammalian cell transfections (~50% cell
death). The square wave pulse provides high mammalian transfection efficiencies and is widely used
for in vivo and in ovo applications. The square wave generates a gentler poration pulse and may not
be as efficient for bacterial transformations as the exponential decay system. The pulse shape can be
altered but is dependent upon the type pulse generator used.

Before Pulse During Pulse After Pulse

Electroporation is a versatile technique that effectively transfects or transforms a large variety of cells
with high efficiencies. The amount of DNA required for electroporation is much lower then for
chemical transfections, resulting in significant savings to the researcher. Our BTX ECM 830 system is
widely used successfully for in vivo, in ovo, ex vivo, and in vitro gene or drug delivery applications.
The BTX ECM 630 system is used most often for transformations of bacterial, yeast, and intact plant

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For more information on these products contact your local VWR sales office,
send an e-mail to vwrbiomarke@eu.vwr.com
Electroporation or visit our website www.vwr.com

Footswitch: Hands-free pulsing. Safety Stand & disposable cuvettes.

Genetrodes: In Vivo gene delivery. Tweezertrodes: In Vivo drug or gene delivery. 2-Needle Array: In Vivo drug or gene
delivery, muscle gene therapy.

with high efficiencies. Both systems can be used with the BTX wide selection of electrodes for in
vivo, in ovo, ex vivo and our BTX HT 96 well systems.

Optimisation of electroporation conditions will also depend on the cell stage prior to electroporation
as well as DNA concentration. The growth rate at which a typical electroporation is performed in
mammalian cell lines is between 50-70% sub-confluence. Mammalian cells, bacteria and yeast
generally need to be in log phase growth for optimal integration of DNA for stable transfections or
transformations as well as expression of molecule in the cytosol for transient expression. The DNA
concentration can vary between mammalian cell types but average DNA concentrations range from
5-40 micrograms/ml for transfections and a range of 0.1 - 10 ng/ml of DNA for transformations.

BTX has designed the BTX 96-Well High Throughput system to quickly run multiple experiments
simultaneously to easily determine the optimal cell and plasmid DNA concentrations as well as
electrical parameters for each application; or run multiple samples for high throughput
electroporation experiments. The HT System boosts productivity by running multiple samples in
seconds, increasing yields and viability.

Electroporation is not only used for transfections but has been successfully adapted to electrofusion
applications. The process of electrofusion uses an AC current which employs low intensity pulse. This
pulse causes a dielectrophoretic alignment current (to form a pearl chain). Once the cell membranes
The fusion Process are in contact, a high intensity electrical field pulse (DC pulse) will create the membrane breakdown
(pores) which merges the two cell membranes together. The AC alignment will be reapplied following
the DC pulse to maintain the cell-cell compression during cell recovery. This results in increased
number of fused hybrids.

Electrofusion is an extremely efficient method for the fusion of mammalian cells. Cell to cell fusion is
a technique used for the production of hybridomas, tetraploid embryos, nuclear transfer embryos
and plant protoplast applications. Electrofusion is quick and easy to perform. Fusion rates range
from 40 to 50% higher, as well as post fusion viability when compared to chemical fusion. In
addition electrofusion is applicable to a wide variety of cell types. It does not require long
incubations, cell handling is much easier in a micro slide, and the electrofusion process is not
cytotoxic to the cells unlike other chemical methods.

HT Plate Handler

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