2.4 Thin Film
Microfluidics
1. Photolithography Aligner (EVG 620).
2. Photoresist (SU8-2015, Microchem).
3. “U-shape” microcap photomask with alignment marker
(TRICR Corporation, CA, USA).
4. 3D NEP chip.
2.5 3D NEP
for Electroporation
1. Bio-Rad Gene X-cell System (Bio-Rad, CA, USA).
2. Copper rod top electrodes (for Subheading 2.2, item 1 and
Subheading 2.2, item 3).
3. ITO glass (for Subheading 2.2, item 2).
4. NEP Chip and assembly (Subheading 2.1, item 2).
2.6 Image
Acquisition and Data
Analysis
1. Fluorescence Microscope: Nikon Eclipse TI.
2. Nikon NIS Elements.
3. Microsoft Excel.
4. Sigma Plot.
3 Methods
3.1 3D NEP System
1. Photoresist SPR200-7 (thickness: 10 μm) with micropore array
is patterned on the other side of silicon chip.
2. The micropore array is etched by DRIE until a deep microwell
array (50 μm in diameter) is formed, connecting with the
nanopore array on the reverse side (Fig. 3a, b).
3. The photoresist SPR950 with an array pattern of nanopores
(650 nm in diameter, pore-to-pore distance: 50 μm), is patterned on a 200 μm thick silicon wafer by projection
photolithography.
4. DRIE is applied to etch the silicon nanopores via SPR950
photoresist (etch depth: 10 μm) (see Note 1).
5. The flat surface with the nanochannel array (Fig. 3c) is
designed for cell loading prior to electroporation.
6. The 3D NEP system consists of four parts, including the silicon
3D NEP chip, a PMMA support platform, PDMS spacers, and
a pair of electrodes (Fig. 1).
7. The substrate, support platform, and two clamps are prefabricated by digital micromilling machine designed by AutoCAD
(see Note 2).
8. A PDMS spacer (thickness: 2 mm) is mounted on the chip,
forming a top chamber where cells are cultured in buffer solution (see Note 2).
3D Nanochannel Array for High-Throughput Cell Manipulation and Electroporation
35
Microfluidics
1. Photolithography Aligner (EVG 620).
2. Photoresist (SU8-2015, Microchem).
3. “U-shape” microcap photomask with alignment marker
(TRICR Corporation, CA, USA).
4. 3D NEP chip.
2.5 3D NEP
for Electroporation
1. Bio-Rad Gene X-cell System (Bio-Rad, CA, USA).
2. Copper rod top electrodes (for Subheading 2.2, item 1 and
Subheading 2.2, item 3).
3. ITO glass (for Subheading 2.2, item 2).
4. NEP Chip and assembly (Subheading 2.1, item 2).
2.6 Image
Acquisition and Data
Analysis
1. Fluorescence Microscope: Nikon Eclipse TI.
2. Nikon NIS Elements.
3. Microsoft Excel.
4. Sigma Plot.
3 Methods
3.1 3D NEP System
1. Photoresist SPR200-7 (thickness: 10 μm) with micropore array
is patterned on the other side of silicon chip.
2. The micropore array is etched by DRIE until a deep microwell
array (50 μm in diameter) is formed, connecting with the
nanopore array on the reverse side (Fig. 3a, b).
3. The photoresist SPR950 with an array pattern of nanopores
(650 nm in diameter, pore-to-pore distance: 50 μm), is patterned on a 200 μm thick silicon wafer by projection
photolithography.
4. DRIE is applied to etch the silicon nanopores via SPR950
photoresist (etch depth: 10 μm) (see Note 1).
5. The flat surface with the nanochannel array (Fig. 3c) is
designed for cell loading prior to electroporation.
6. The 3D NEP system consists of four parts, including the silicon
3D NEP chip, a PMMA support platform, PDMS spacers, and
a pair of electrodes (Fig. 1).
7. The substrate, support platform, and two clamps are prefabricated by digital micromilling machine designed by AutoCAD
(see Note 2).
8. A PDMS spacer (thickness: 2 mm) is mounted on the chip,
forming a top chamber where cells are cultured in buffer solution (see Note 2).
3D Nanochannel Array for High-Throughput Cell Manipulation and Electroporation
35