milling machine. The centrifugal holder is made by a PMMA plate
of ½
00 thickness. The centrifugal holder is designed as axial symmetrical to balance the centrifugal force.
3.3 Centrifugal Cell
Trapping and
Electroporation
Biochips and cover PDMS is first assembled and placed inside the
PMMA case. 200–300 cells in 5 μL suspension are injected into the
cell side inlet of the biochip by using a pipette. The assembled and
cell loaded biochip is placed in the centrifugal holder and placed in a
centrifuge. After spinning for 3 min at 45 Â g, the PMMA case is
removed from the centrifugal holder. Plasmid or nanoparticle
encapsulated plasmid suspension is then loaded into the DNA
side inlet slowly to avoid pushing away the trapped cells. Two
palladium wires are then inserted into the cell side inlet and DNA
side inlet, respectively. A Bio-Rad Gene Pulser Xcell electroporation
system is then connected to the two wires. A voltage pulse sequence
is then used depending on the dimension of the nanochannel and
the microchannel, and types of plasmids. A typical voltage pulse
sequence is 250 V, 10 ms pulse length, and five pulses.
References
1. Chang L et al (2016) MicroÀ/nanoscale electroporation. Lab Chip 16(21):4047–4062
2. Chang L et al (2015) Magnetic tweezers-based
3D microchannel electroporation for highthroughput gene transfection in living cells.
Small 11(15):1818–1828
3. Boukany PE et al (2011) Nanochannel electroporation delivers precise amounts of biomolecules into living cells. Nat Nanotechnol 6
(11):747–754
4. Andresen KØ et al (2010) Injection molded
chips with integrated conducting polymer electrodes for electroporation of cells. J Micromech
Microeng 20(5):055010
5. Zhao X et al (2017) Laser micromachining of
reusable glass devices dedicated to the targeted
electroporation of cell assemblies. In: 2017 19th
international conference on solid-state sensors,
actuators and microsystems (TRANSDUCERS),
Kaohsiung, Taiwan, pp 1608–1611
6. Yuan H, Cambron SD, Crain MM, Keynton RS
(2016) Fabrication of a micro/nanofluidic platform via three-axis robotic dispensing system.
J Micro Nano-Manuf 4(4):041005
7. Gao K et al (2014) Design of a microchannelnanochannel-microchannel array based nanoelectroporation system for precise gene transfection. Small 10(5):1015–1023
8. SU-8 2000 Data Sheet. MicroChem
Micromachining of Polymeric Microfluidic Micro/Nanoelectroporation Device
27
of ½
00 thickness. The centrifugal holder is designed as axial symmetrical to balance the centrifugal force.
3.3 Centrifugal Cell
Trapping and
Electroporation
Biochips and cover PDMS is first assembled and placed inside the
PMMA case. 200–300 cells in 5 μL suspension are injected into the
cell side inlet of the biochip by using a pipette. The assembled and
cell loaded biochip is placed in the centrifugal holder and placed in a
centrifuge. After spinning for 3 min at 45 Â g, the PMMA case is
removed from the centrifugal holder. Plasmid or nanoparticle
encapsulated plasmid suspension is then loaded into the DNA
side inlet slowly to avoid pushing away the trapped cells. Two
palladium wires are then inserted into the cell side inlet and DNA
side inlet, respectively. A Bio-Rad Gene Pulser Xcell electroporation
system is then connected to the two wires. A voltage pulse sequence
is then used depending on the dimension of the nanochannel and
the microchannel, and types of plasmids. A typical voltage pulse
sequence is 250 V, 10 ms pulse length, and five pulses.
References
1. Chang L et al (2016) MicroÀ/nanoscale electroporation. Lab Chip 16(21):4047–4062
2. Chang L et al (2015) Magnetic tweezers-based
3D microchannel electroporation for highthroughput gene transfection in living cells.
Small 11(15):1818–1828
3. Boukany PE et al (2011) Nanochannel electroporation delivers precise amounts of biomolecules into living cells. Nat Nanotechnol 6
(11):747–754
4. Andresen KØ et al (2010) Injection molded
chips with integrated conducting polymer electrodes for electroporation of cells. J Micromech
Microeng 20(5):055010
5. Zhao X et al (2017) Laser micromachining of
reusable glass devices dedicated to the targeted
electroporation of cell assemblies. In: 2017 19th
international conference on solid-state sensors,
actuators and microsystems (TRANSDUCERS),
Kaohsiung, Taiwan, pp 1608–1611
6. Yuan H, Cambron SD, Crain MM, Keynton RS
(2016) Fabrication of a micro/nanofluidic platform via three-axis robotic dispensing system.
J Micro Nano-Manuf 4(4):041005
7. Gao K et al (2014) Design of a microchannelnanochannel-microchannel array based nanoelectroporation system for precise gene transfection. Small 10(5):1015–1023
8. SU-8 2000 Data Sheet. MicroChem
Micromachining of Polymeric Microfluidic Micro/Nanoelectroporation Device
27