Dielectrophoretic Manipulation of Cell Transfection Efficiency during Electroporation Using a Center Needle Electrode

Long duration electric pulses are frequently used to facilitate DNA electrotransfer into cells and tissues, while electroporation pulses can be combined with electrophoresis to maximize the transfection efficiency. In this work, we present the dielectrophoresis (DEP)-assisted methodology for electrotransfer of plasmid DNA (3.5 kbp pmaxGFP) into mammalian cells (CHO-K1). A prototype of an electroporation cuvette with center needle electrode for DEP-assisted transfection is presented resulting in a 1.4-fold of transfection efficiency increase compared to the electroporation-only procedure (1.4 kV/cm x 100 mu s x 8). The efficiency of transfection has been compared between three DEP frequencies of 1, 100, and 1 MHz. Lastly, the effects of exposure time (1, 3, and 5 min) during the DEP application step have been determined. It is concluded that the proposed methodology and exposure setup allow a significant improvement of transfection efficiency and could be used as an alternative to the currently popular electrotransfection techniques.

Automated summary

In this study, a dielectrophoresis-assisted methodology for electrotransfer of plasmid DNA into mammalian cells was developed, resulting in a significant increase in transfection efficiency compared to traditional electroporation-only procedures. The use of a prototype electroporation cuvette with a center needle electrode for DEP-assisted transfection showed a 1.4-fold improvement in efficiency. The effects of different DEP frequencies and exposure times on transfection efficiency were investigated, highlighting the potential of this method as an alternative to current electrotransfection techniques.