Chapter 4
3D Nanochannel Array for High-Throughput Cell
Manipulation and Electroporation
Lingqian Chang, Stephen Black, Chandani Chitrakar, and Mehdi Nouri
Abstract
Electroporation has been one of the most commonly used physical methods for gene/drug delivery.
Compared to other nonviral counterparts, electroporation enables optimization of delivery efficiency by
tuning the electric field applied on cells. Commercial electroporation, however, results in stochastic
transfection and significant cellular damage mostly due to its “bulk” environment. In this chapter, we
introduce nanoelectroporation (NEP) which has demonstrated living cell transfection in a highly controllable manner. In NEP, the electric field can be precisely focused on a single cell positioned on nanochannels.
Safe single-cell electroporation as well as “electrophoretic” molecular delivery can be achieved on the same
device. This system achieves significantly higher transfection efficiency and cellular viability than commercial systems. This device is unique in that it can efficiently deliver genetic molecules (e.g., DNAs, RNAs)
that exceed 10 kbp in size. The NEP device based on a 3D nanochannel array prototype was fabricated
using cleanroom techniques. For achieving precise cell to nanochannel pairing, three on-chip highthroughput manipulation technologies were developed, that is, magnetic tweezers (MT), dielectrophoresis
(DEP), and thin-film microfluidics.
Key words Nanoelectroporation, Nanochannel array, Magnetic tweezers, Dielectrophoresis,
Microfluidics
1 Introduction
A wide variety of methods [1] have been developed for introducing
exogenous materials into cells. These include viral, chemical (lipofectamine, polyplex, etc.), and physical (gene gun [2, 3], microinjection [4], laser ablation [5, 6], electroporation [7, 8], etc.)
methods. Transfection using viruses is highly efficient, but safety
is a concern with what else the virus may do [9]. As for nonviral
methods, chemical methods generally transfect cells by tricking the
cells into performing endocytosis. This is where the membrane of
the cell will pinch a bubble of membrane into the cell. These
methods are efficient to deliver drugs in mass quantity to the
body, but current technologies results in a slow, stochastic delivery
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_4,
© Springer Science+Business Media, LLC, part of Springer Nature 2020
29
3D Nanochannel Array for High-Throughput Cell
Manipulation and Electroporation
Lingqian Chang, Stephen Black, Chandani Chitrakar, and Mehdi Nouri
Abstract
Electroporation has been one of the most commonly used physical methods for gene/drug delivery.
Compared to other nonviral counterparts, electroporation enables optimization of delivery efficiency by
tuning the electric field applied on cells. Commercial electroporation, however, results in stochastic
transfection and significant cellular damage mostly due to its “bulk” environment. In this chapter, we
introduce nanoelectroporation (NEP) which has demonstrated living cell transfection in a highly controllable manner. In NEP, the electric field can be precisely focused on a single cell positioned on nanochannels.
Safe single-cell electroporation as well as “electrophoretic” molecular delivery can be achieved on the same
device. This system achieves significantly higher transfection efficiency and cellular viability than commercial systems. This device is unique in that it can efficiently deliver genetic molecules (e.g., DNAs, RNAs)
that exceed 10 kbp in size. The NEP device based on a 3D nanochannel array prototype was fabricated
using cleanroom techniques. For achieving precise cell to nanochannel pairing, three on-chip highthroughput manipulation technologies were developed, that is, magnetic tweezers (MT), dielectrophoresis
(DEP), and thin-film microfluidics.
Key words Nanoelectroporation, Nanochannel array, Magnetic tweezers, Dielectrophoresis,
Microfluidics
1 Introduction
A wide variety of methods [1] have been developed for introducing
exogenous materials into cells. These include viral, chemical (lipofectamine, polyplex, etc.), and physical (gene gun [2, 3], microinjection [4], laser ablation [5, 6], electroporation [7, 8], etc.)
methods. Transfection using viruses is highly efficient, but safety
is a concern with what else the virus may do [9]. As for nonviral
methods, chemical methods generally transfect cells by tricking the
cells into performing endocytosis. This is where the membrane of
the cell will pinch a bubble of membrane into the cell. These
methods are efficient to deliver drugs in mass quantity to the
body, but current technologies results in a slow, stochastic delivery
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_4,
© Springer Science+Business Media, LLC, part of Springer Nature 2020
29