Chapter 3
Micromachining of Polymeric Microfluidic
Micro/Nanoelectroporation Device
Lei Li
Abstract
Micro/nanochannel electroporation can deliver gene/drug into single cell with precise dosage control and
much higher cell viability compared to traditional bulk electroporation. However, single cell micro/
nanochannel electroporation has the problems of low efficiency and complicated operation. By integrating
microfluidic with micro/nanochannel electroporation, a large number of cells can be processed within a
short time. In this chapter, we provide a detailed protocol of fabrication microfluidic nanochannel electroporation devices. The fabrication of this microfluidic nanochannel electroporation device integrates soft
lithography, DNA combing and imprinting, and micromilling. This device is appropriate for gene/drug
delivery to a batch of cells. It has the advantages of both the single cell nanochannel electroporation and
microfluidic based cell manipulation. The procedures of device fabrication, holder fabrication, cell trapping,
and electroporation are included in this protocol.
Key words Nanochannel electroporation, Micromachining, Soft lithography, DNA combing and
imprinting, Microfluidics, Centrifugal force cell trapping
1 Introduction
Single cell microelectroporation (MEP) has been used to delivery
gene/drug into cells. By placing a cell next to a microaperture or a
microchannel, focused electric fields can create temporary pores in
cell membranes. Furthermore, nanochannel electroporation (NEP)
technology has been developed recently. NEP technology can
directly deliver biomolecules into the cell cytosol across nanochannels with highly concentrated electrical fields and much smaller
generated pores on cell membranes [1]. Micro/nanoelectroporation devices have been reported to have superiorities of lower
poration voltages, better transfection efficiency, and lower cell
mortality compared to conventional bulk electroporation (BEP),
especially for single cell analysis.
MEP/NEP is a highly effective tool to directly delivery gene/
drug into single living cells. However, current MEP/NEP devices
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_3,
© Springer Science+Business Media, LLC, part of Springer Nature 2020
21
Micromachining of Polymeric Microfluidic
Micro/Nanoelectroporation Device
Lei Li
Abstract
Micro/nanochannel electroporation can deliver gene/drug into single cell with precise dosage control and
much higher cell viability compared to traditional bulk electroporation. However, single cell micro/
nanochannel electroporation has the problems of low efficiency and complicated operation. By integrating
microfluidic with micro/nanochannel electroporation, a large number of cells can be processed within a
short time. In this chapter, we provide a detailed protocol of fabrication microfluidic nanochannel electroporation devices. The fabrication of this microfluidic nanochannel electroporation device integrates soft
lithography, DNA combing and imprinting, and micromilling. This device is appropriate for gene/drug
delivery to a batch of cells. It has the advantages of both the single cell nanochannel electroporation and
microfluidic based cell manipulation. The procedures of device fabrication, holder fabrication, cell trapping,
and electroporation are included in this protocol.
Key words Nanochannel electroporation, Micromachining, Soft lithography, DNA combing and
imprinting, Microfluidics, Centrifugal force cell trapping
1 Introduction
Single cell microelectroporation (MEP) has been used to delivery
gene/drug into cells. By placing a cell next to a microaperture or a
microchannel, focused electric fields can create temporary pores in
cell membranes. Furthermore, nanochannel electroporation (NEP)
technology has been developed recently. NEP technology can
directly deliver biomolecules into the cell cytosol across nanochannels with highly concentrated electrical fields and much smaller
generated pores on cell membranes [1]. Micro/nanoelectroporation devices have been reported to have superiorities of lower
poration voltages, better transfection efficiency, and lower cell
mortality compared to conventional bulk electroporation (BEP),
especially for single cell analysis.
MEP/NEP is a highly effective tool to directly delivery gene/
drug into single living cells. However, current MEP/NEP devices
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_3,
© Springer Science+Business Media, LLC, part of Springer Nature 2020
21