Development of Device for Electroporation of Cells – Niels Bohr Institute - University of Copenhagen

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Development of Device for Electroporation of Cells

Electroporation of biological cells is a mature and widely employed tool which still holds a lot of promise for novel applications in the future. Allowing for both the transfection of impermeant exogenous material through the cell membrane and for the release of intracellular content, the areas of realized and potential application include genetic manipulation, cloning, cell fusion, cytometry, chromosomal mapping, antibody production, drug screening, and cancer immune therapy.

This project describes the design and development of a microfluidic cell electroporation device in which the cell medium itself acts as electrodes, employing a channel with a constriction which serves to enhance the electrical field in the narrow section such that the field there is strong enough for electroporation and the field in the rest of the channel is too weak to have an adverse effect on the viability of cells. This method avoids the high voltages associated with pulse generators, and avoids the difficulties associated with microfabricated electrodes, namely inhomogeneous fields resulting from minor faults in the electrode geometry, and electrode deterioration. In addition, the flow-through system allows for the fast transfer of heat away from the channel constriction, and has potential for integration into microfluidic systems with multiple functions.

The project details a complete production cycle, from lithographically defining the channel structure in photoresist on a silicium wafer in the clean-room at Risø, forming the master for a nikkel mold, mass-producing the two halves of the device in the injection-molder at Risø, lithographically defining the electrodes in conducting polymer, thermal bonding, and finally, characterizing the properties of the device and confirming functionality by transfecting cells from the THP-1 monocyte cell line with a fluorescent dye, taking care to document that cells are alive following electroporation.

Master thesis defence by Kristian Andresen