Chapter 5
Low-Voltage Flow-Through Electroporation
Membrane and Method
Juliette Experton, Aaron G. Wilson, and Charles R. Martin
Abstract
Electroporation uses high electric field gradients to create pores within the membrane of living cells in order
to deliver a substance, for example a gene, into the cytoplasm. To achieve such gradients, current
electroporation devices deliver voltage pulses in the kV range to the cell medium. We describe here a new
device based on gold-microtube membranes that can accomplish electroporation with voltage pulses that
are orders of magnitude smaller, 4 V. The percentages of electroporated bacteria were found to be more
than an order of magnitude higher than obtained with a commercial electroporator.
Key words Electroporation, Gold-microtube membranes, Escherichia coli, Electric field gradient
1 Introduction
When a living cell is placed in a solution containing a large electric
field gradient, the voltage difference across the cell membrane can
become high enough that pores form in the membrane—electroporation [1–3]. Electroporation is a widely used technology for
introducing exogeneous substances, for example genes and molecular probes, into cells [4–6]. Depending on the application, electroporation may be reversible, meaning the pores close after some
time, or irreversible, which results in cell death.
Currently available electroporation devices use voltage pulses in
the kV range in order to achieve electric field gradients across the
cell membrane high enough to porate the cell, for example
3.65 kV/cm for Escherichia coli [7]. Such large voltage pulses are
undesirable because they require special safety precautions,
complicated experimental procedures, and produce high cell
mortality [8–10].
We have developed a new microscale electroporation device
that uses a commercially available membrane filter with gold
microtubes deposited in the pores [11]. Because the tubes are
Shulin Li et al. (eds.), Electroporation Protocols: Microorganism, Mammalian System, and Nanodevice,
Methods in Molecular Biology, vol. 2050, https://doi.org/10.1007/978-1-4939-9740-4_5,
© Springer Science+Business Media, LLC, part of Springer Nature 2020
43
Low-Voltage Flow-Through Electroporation
Membrane and Method
Juliette Experton, Aaron G. Wilson, and Charles R. Martin
Abstract
Electroporation uses high electric field gradients to create pores within the membrane of living cells in order
to deliver a substance, for example a gene, into the cytoplasm. To achieve such gradients, current
electroporation devices deliver voltage pulses in the kV range to the cell medium. We describe here a new
device based on gold-microtube membranes that can accomplish electroporation with voltage pulses that
are orders of magnitude smaller, 4 V. The percentages of electroporated bacteria were found to be more
than an order of magnitude higher than obtained with a commercial electroporator.
Key words Electroporation, Gold-microtube membranes, Escherichia coli, Electric field gradient
1 Introduction
When a living cell is placed in a solution containing a large electric
field gradient, the voltage difference across the cell membrane can
become high enough that pores form in the membrane—electroporation [1–3]. Electroporation is a widely used technology for
introducing exogeneous substances, for example genes and molecular probes, into cells [4–6]. Depending on the application, electroporation may be reversible, meaning the pores close after some
time, or irreversible, which results in cell death.
Currently available electroporation devices use voltage pulses in
the kV range in order to achieve electric field gradients across the
cell membrane high enough to porate the cell, for example
3.65 kV/cm for Escherichia coli [7]. Such large voltage pulses are
undesirable because they require special safety precautions,
complicated experimental procedures, and produce high cell
mortality [8–10].
We have developed a new microscale electroporation device
that uses a commercially available membrane filter with gold
microtubes deposited in the pores [11]. Because the tubes are
Shulin Li et al. (eds.), Electroporation Protocols: Microorganism, Mammalian System, and Nanodevice,
Methods in Molecular Biology, vol. 2050, https://doi.org/10.1007/978-1-4939-9740-4_5,
© Springer Science+Business Media, LLC, part of Springer Nature 2020
43