Nucleofection
cells were transfected using the Amaxa nucleofector II device (Lonza).
Cells were re suspended in specific solutions. Briefly, according to
amaxa guidelines ,2.5×\(10^{6}\) RPMI-8226 cells were re suspended in
100 μl of Nucleofection buffer solution containing 5μg DNA, and the
cells were transfected using Program G-016 of the Nucleofector II and
immediately transferred into 6-well plates containing 37 °C pre-warmed
culture medium .
Device Design and FabricationMold was fabricated using conventional photolithography techniques. SU-8
2550 negative photoresist was spin coated on silicon wafer and baked.
PDMS pre-polymer and the cross linker were mixed at the recommended 10:1
w/w ratio. After pouring the mixture onto the SU-8 master and degassing
for 1 h, the mixture was cured at 75 C for 2 h. PDMS moldings were cut
and peeled off the masters. The main components of such devices are: (i)
inlet and outlet (ii) main channel: a serpentine-like channel, 100µm
width and 60 µm height, consisting of turns with length of 12cm.