Electroporation (EP) is one of the most significant biotechnologies, which can be used for drug delivery, DNA transfection, and cell lysis due to its high efficiency and safety. To date, micro/nanotechnology has already been used for electroporation (EP). The aluminum nano-spike EP (ANS-EP) chip has been successfully fabricated for the delivery of biomolecules into cells at low voltage. However, due to the material limitation, the nano-spike is highly deformable and highly chemically reactive.

This thesis mainly focuses on the systematic design and fabrication of silicon nano-pillar electroporation chips for the delivery of molecules using projection photolithography to improve the mechanical property of EP chips based on the nano-electric field amplification. Numerical simulat...[ Read more ]

Electroporation (EP) is one of the most significant biotechnologies, which can be used for drug delivery, DNA transfection, and cell lysis due to its high efficiency and safety. To date, micro/nanotechnology has already been used for electroporation (EP). The aluminum nano-spike EP (ANS-EP) chip has been successfully fabricated for the delivery of biomolecules into cells at low voltage. However, due to the material limitation, the nano-spike is highly deformable and highly chemically reactive.

This thesis mainly focuses on the systematic design and fabrication of silicon nano-pillar electroporation chips for the delivery of molecules using projection photolithography to improve the mechanical property of EP chips based on the nano-electric field amplification. Numerical simulations of electric field strength indicated that enhanced localized electric field at the tip based on the high-aspect-ratio of nano-pillars and show the feasibility of using nano-pillar in the subsequent chip. Arrays of Au-SiNP-EP chips were fabricated through ASML stepper, DRIE, AOE, and ARC sputter processes.

A serial of biological experiments has been designed to analyze the effect of different electric pulse parameters using Hela cells with digital fluorescent microscopy with Acridine Orange (AO) and Propidium Iodide (PI) dyes. The electric field pulse parameters have been optimized to get electric pulse protocols, which can deliver intracellular molecules with high EP efficiency ƞ_EP (~80%) and cell viabilities ф_cell (~80%) at low pulse amplitudes V_a (4V) and low durations time t_p (2ms) successfully. Critical values for electroporation were determined for different electric pulse parameters and the phase diagram of the different pulse duration time has been accomplished, which can guide the subsequent experiments with other cells.

What is more, the Arduino based mini-Electroporator for my experiment is successfully designed. The principle of Arduino based signal generation-electroporator using the circuit diagram. Besides, the protocol of the electroporator has been optimized to achieve a low pulse range. By changing the code from Arduino, the user can minimize the experiment process. In summary, there are three advantages over traditional electroporators. Firstly, based on the nanopillar, it can introduce the EP under the lower amplitude pulse, which can improve the safety of the equipment. Then, the design of the electroporator is light and portable. Two parts of the shell have been designed using SOLIDWORKS and fabricated by the 3 D printer and laser cut. Users can take it to the experimental laboratory, which can reduce the contamination problems because it is not convenient for bulk electroporator. Last but not least, the protocol is controlled by an Arduino, the user can minimize the experiment process and different protocols can be designed based on the formal study.