(516b) Optimizing Membrane Sandwich Electroporation Through Microfabrication And High Speed Spin Disk Confocal Imaging

Electroporation is a common technology for delivering genes and small or large molecules into cells. Due to its physical nature electroporation does not incur immune response or toxicity like virus and artificial virus vectors. Ideally 100% transfection efficiency and cell viability are desired, however a user has to make trade offs between these results with bulk electroporation. Recently research labs have developed local electroporation. This process enhances electroporation results by using holes in a dielectric membrane to focus the electric field and to localize the site where a cell is electroporated. Local electroporation has shown significant enhancements over bulk electroporation.

We have developed a new generation of electroporation technology called membrane sandwich electroporation (MSE). MSE uses two dielectric porous membranes to enhance electroporation. Pores in the membranes focus the electric field to create a site of electroporation on a cell and concentrate DNA near the site of electroporation. Results with NIH-3T3 cells and pEGFP or pSEAP have shown improvements over bulk and local electroporation.

We are currently studying the mechanisms of DNA concentration and entry as well as parameters involved in MSE to optimize this process. We have fabricated a micro device and an array of micro-pores in a membrane to accomplish this goal. This new array membrane will replace the current membrane that has random pore locations. A well defined array will ensure each cell experiences the same conditions during MSE while parameters are optimized. In addition to the new membrane we will be visualizing the electroporation process using a high speed spin-disk confocal microscope to investigate mechanisms involved in MSE.