Description:

A dielectrophoresis based system has been developed to separate individual cells of interest, such as the cells over expressing a protein of interest that is tagged to the green fluorescent protein(GFP). Dielectrophoresis technologies allow us to manipulate cells in small sample volumes with a good amount of control and accuracy. Here we propose the use of micro capillary based electrodes for the purpose of separating the cells which have incorporated the GFP constructs from the non-expressing cells. This electrode design generates a highly non-uniform electric field around the tip, thereby providing a method to relocate a single cell. Electric field simulations have been used to optimize the electrode geometry and also to determine a suitable working voltage. These electrodes are fitted on to micromanipulator stages, which allow us to precisely control the placement of cells. The fluorescence based excitation of the GFP marker, will be used to identify the cells to be picked up. These cells can then be deposted into a microwell and the selected cells can be extracted from the well, leaving us with an enriched population of clonal cells.

This method allows us to bypass the need for a large cell population to be sorted via conventional FACS analysis. This technology will help us achieve early isolation of certain gene specific cells and it also offers a more cost effective alternative to conventional sorting methods. Thus we propose a new method of achieving highly targetted isolation of transfected cell lines.