tin oxide (ITO) and is small enough to fit inside of a petri dish for
cell culturing. A conductive and transparent lower electrode consisting of a glass slide coated in ITO serves as the foundation of
the chip. A layer of PDMS containing microfluidic channels for
the material to be delivered is bonded to the conductive surface of
the glass slide. The microfluidic channels protect the cells from
fluid flow and the resulting shear stresses which can be damaging
to the cells [12]. A perforated polycarbonate (PC) surface is
placed above the PDMS followed by a second layer of PDMS
which acts as the cell culture chamber. This chapter discusses
delivery of green fluorescent protein (GFP) plasmid into HeLa
or HT1080 cells but the same chip has also been used to transfect
differentiated neural stem cells [9].
2 Materials
2.1 Microdevice
Fabrication and
Assembly
1. Indium-tin oxide (ITO) glass slide, MilliporeSigma, USA.
2. Wafer.
3. Optical microscope.
4. Polydimethylsiloxane (PDMS).
5. Oxygen plasma machine.
6. Oven.
7. Pluronic F-127 (poloxamer).
8. Phosphate buffered saline (PBS).
9. Polycarbonate (PC) membrane, AR Brown, USA (see Note 1).
2.2 Cell Culture
1. HeLa cells (ATCC CCL-2), American Type Culture
Collection, USA.
2. Dulbecco’s
Modified
Eagle’s
medium
(DMEM),
MilliporeSigma, USA.
3. pH indicators: L-glutamine and phenol red.
4. Fetal bovine serum (FBS), MilliporeSigma, USA.
5. 1Â penicillin–streptomycin, MilliporeSigma, USA.
2.3 Plasmid DNA,
Stains, and Dye
1. 2 MDa green fluorescent protein (GFP) plasmid.
2. Hoechst 33342 stain, Invitrogen, USA.
3. Propidium iodide (PI) stain, eBioscience, USA.
4. Calcein acetoxymethylester (AM) dye, Invitrogen, USA.
2.4 Electroporation
1. Function generator: DS345, Stanford Research Systems, USA
(see Note 2).
2. Voltage amplifier: OPA445, Texas Instruments, USA.
Microfluidic Device for Localized Electroporation
93
cell culturing. A conductive and transparent lower electrode consisting of a glass slide coated in ITO serves as the foundation of
the chip. A layer of PDMS containing microfluidic channels for
the material to be delivered is bonded to the conductive surface of
the glass slide. The microfluidic channels protect the cells from
fluid flow and the resulting shear stresses which can be damaging
to the cells [12]. A perforated polycarbonate (PC) surface is
placed above the PDMS followed by a second layer of PDMS
which acts as the cell culture chamber. This chapter discusses
delivery of green fluorescent protein (GFP) plasmid into HeLa
or HT1080 cells but the same chip has also been used to transfect
differentiated neural stem cells [9].
2 Materials
2.1 Microdevice
Fabrication and
Assembly
1. Indium-tin oxide (ITO) glass slide, MilliporeSigma, USA.
2. Wafer.
3. Optical microscope.
4. Polydimethylsiloxane (PDMS).
5. Oxygen plasma machine.
6. Oven.
7. Pluronic F-127 (poloxamer).
8. Phosphate buffered saline (PBS).
9. Polycarbonate (PC) membrane, AR Brown, USA (see Note 1).
2.2 Cell Culture
1. HeLa cells (ATCC CCL-2), American Type Culture
Collection, USA.
2. Dulbecco’s
Modified
Eagle’s
medium
(DMEM),
MilliporeSigma, USA.
3. pH indicators: L-glutamine and phenol red.
4. Fetal bovine serum (FBS), MilliporeSigma, USA.
5. 1Â penicillin–streptomycin, MilliporeSigma, USA.
2.3 Plasmid DNA,
Stains, and Dye
1. 2 MDa green fluorescent protein (GFP) plasmid.
2. Hoechst 33342 stain, Invitrogen, USA.
3. Propidium iodide (PI) stain, eBioscience, USA.
4. Calcein acetoxymethylester (AM) dye, Invitrogen, USA.
2.4 Electroporation
1. Function generator: DS345, Stanford Research Systems, USA
(see Note 2).
2. Voltage amplifier: OPA445, Texas Instruments, USA.
Microfluidic Device for Localized Electroporation
93