Quantified Femtosecond Laser Based Opto-perforation of Living Cells

Opto-perforation is an interesting alternative to conventional techniques for gene transfer into living cells. The cell membrane is perforated by femtosecond (fs) laser pulses, in order to induce an uptake of macromolecules. In this study, we successfully transfected cells with GFP vector. Transfected cells were not showing any signs of apoptosis or necrosis after 48 h. Based on simultaneously measured membrane potential changes, we calculated and experimentally verified that the relative volume exchanged is 0.4 times the total cell volume. Thus, for the first time quantitative predication of the amount of uptaken molecules and therefore a quantification of transfection is possible.