Chapter 12
Targeted In Vivo Electroporation Using Nanoengineered
Microelectrodes
Daniel Schwarz and Andreas T. Schaefer
Abstract
Targeted electroporation by using glass microelectrodes is a popular and versatile tool allowing for easy
manipulation of single cells and cell ensembles in living tissue. Because of the highly focal distribution of the
electric field, however, the range of reversible electroporation without causing irreversible damage is tight—
especially when aiming for larger electroporation volumes. In this chapter, we describe the production of
nanoengineered electroporation microelectrodes (NEMs), a practicable way to prepare glass microelectrodes providing a more even distribution around the tip of a pipette by using nanotechnological methods.
Key words Electroporation, Nanoengineering, Microelectrodes, In vivo
1 Introduction
The technique of targeted electroporation using a glass microelectrode was popularized by Haas et al. in 2001 to transfect single
neuronal cells of the tadpole in vivo [1]. They employed fine needle
glass microelectrodes as normally used for patch-clamp recordings
to deliver the electric field. This versatile approach greatly broadened the in vivo applicability of electroporation because it proved
that highly inhomogeneous electric fields with a distant grounding
electrode could effectively be employed for the purpose and that
plate or array electrodes were not necessarily required. In
subsequent years, this approach has increasingly been used especially in the neurosciences and has also been shown to be effective
beyond the single cell level [2–5].
An important limitation, however, comes with the highly focal
electric field distribution of such glass microelectrodes: Because of
the small pipette tip of ~1 μm in diameter, the electric peak potentials centered to the tip rise rapidly with increasing stimulation
intensity of the system [6]. This is a major drawback when the
technique is intended for larger electroporation volumes as
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_12,
© Springer Science+Business Media, LLC, part of Springer Nature 2020
113
Targeted In Vivo Electroporation Using Nanoengineered
Microelectrodes
Daniel Schwarz and Andreas T. Schaefer
Abstract
Targeted electroporation by using glass microelectrodes is a popular and versatile tool allowing for easy
manipulation of single cells and cell ensembles in living tissue. Because of the highly focal distribution of the
electric field, however, the range of reversible electroporation without causing irreversible damage is tight—
especially when aiming for larger electroporation volumes. In this chapter, we describe the production of
nanoengineered electroporation microelectrodes (NEMs), a practicable way to prepare glass microelectrodes providing a more even distribution around the tip of a pipette by using nanotechnological methods.
Key words Electroporation, Nanoengineering, Microelectrodes, In vivo
1 Introduction
The technique of targeted electroporation using a glass microelectrode was popularized by Haas et al. in 2001 to transfect single
neuronal cells of the tadpole in vivo [1]. They employed fine needle
glass microelectrodes as normally used for patch-clamp recordings
to deliver the electric field. This versatile approach greatly broadened the in vivo applicability of electroporation because it proved
that highly inhomogeneous electric fields with a distant grounding
electrode could effectively be employed for the purpose and that
plate or array electrodes were not necessarily required. In
subsequent years, this approach has increasingly been used especially in the neurosciences and has also been shown to be effective
beyond the single cell level [2–5].
An important limitation, however, comes with the highly focal
electric field distribution of such glass microelectrodes: Because of
the small pipette tip of ~1 μm in diameter, the electric peak potentials centered to the tip rise rapidly with increasing stimulation
intensity of the system [6]. This is a major drawback when the
technique is intended for larger electroporation volumes as
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_12,
© Springer Science+Business Media, LLC, part of Springer Nature 2020
113