Abstract
Cells play significant role in our day to day life. However the interactions of cells to cells responses with their organelles to molecules and intracellular behavior are still not fully understood. To understand better physiological interactions among molecules, organelles, and cells ensemble average measurement for millions of cells together sometimes cannot provide the detail information. For example, the differentiation behavior of stem cells or metastasis process of tumor initiation cells. Thus, single cell level research is becoming a pioneering research area that unveils the interaction details in high temporal and spatial resolution among cells. To analyze the intracellular biochemical function, single cell electroporation can be conducted by employing a miniaturized device, whose dimension should be similar to that of a single cell. Micro/nanofluidic devices can fulfill this requirement for single cell electroporation. This device is not only useful for cell lysis, cell to cell fusion or separation, insertion of drug, DNA and antibodies inside single cell, but also it can control biochemical, electrical and mechanical parameters using electroporation technique. This device provides better performance such as high transfection efficiency, high cell viability, lower Joule heating effect, less sample contamination, lower toxicity during electroporation experiment when compared to bulk measurement. In most recent years, single organelles within a cell can be analyzed selectively by reducing the electrode size and gap at nanoscale level. This advanced technique can deliver (in/out) biomolecules precisely through a small membrane area (micro to nanoscale area) of the single cell, known as localized single cell nano-electroporation (LSCNEP). This thesis will provide an overview of bimolecular delivery by nano-electroporation technique into different single cell such as human cervical cancer (HeLa), human colon cancer (HCT-8), human gastric cancer (AGS) and human lung cancer (Cl1(0) cells. The delivery was performed through multi-nanolocalize single cell membrane regions with their manipulation, transfection, lysis and dynamic analysis. This nano-fluidic chip was fabricated with array of Indium Tin Oxide (ITO) based transparent triangular 40 nm metal nano-electrode tip with 25 nm to 70 nm gap between two nano-electrodes, resulting an intense electric field in-between nano-electrode gap. As results, only a small regions (nanolocalize) on different area of single cell affected to deliver biomolecules form outside to inside of the cell, where reaming other area of the membrane was unaffected. Moreover, we deposited a high dielectric passivation layer on top of the nano-electrode chip to reduce thermal and ionic effect during experiment. This novel device with multi-nanolocalized nano-electroporation technique successfully reduces cell toxicity, power consumption, hydrogen and hydroxyl ion generation, bubble formation during electroporation experiment resulting to increase the high transfection efficiency with high cell viability. The device has spatial, temporal and qualitative dosage control, which potentially applicable for medical therapeutics and biological cell studies.