Electroporation microchips for continuous gene transfection

Electroporation is a technique with which DNA molecules can be delivered into cells in a chamber using high electric field pulses. Compared to the commercial equipment, the fabricated flow-type electroporation microchip overcomes the limit in the amount of target cells and the potential risk of using high voltage, which are the two drawbacks in current electroporation technology. The flowing mechanism eliminates the temperature-rising effect found in a commercial sterile plastic cuvette during electroporation and thus can improve the survival rate of cells after electroporation. ectroporation microchip, consisting of a micro-channel with gold thin film electrodes on both sides, was fabricated on PMMA material using evaporation, photolithography, lift-off and fusion-bonding methods. The suspension fluid of Huh-7 cell lines mixed with reporter genes flowed through the channel under a series of electrical square pulses. Trypan blue staining was used to assess the viability of the cells treated with pulses. The cells were evaluated under a microscope. mensions of the channel were 5 mm wide, 0.2 mm high and 25 mm long. Cell experiments demonstrated that our microchip could transfect the flowing cells with the expression of reporter genes with lower dissipated heat. The survival rate increased with a lower pulse frequency and a higher flow speed. Our newly designed novel microchip for electroporation of continuous cell flow will have great potential for future gene delivery in hematologic malignancy.