The irradiation of cells combined with the immunoconjugate of gold nanoparticles by the short pulse laser can make the plasma membrane be transiently permeabilized, which can be used to transfer exogenous molecules into the cells. We explore this technique as a novel gene transfection method for floating cells. Three different floating cells exposed to the laser are selectively transfected with fluorescein isothiocyanatedextran, antibody, and green fluorescent protein (GFP) coding plasmids, and the viability of cells are determined by propidium iodide. For fluorescein isothiocyanate-dextran, the best transfection efficiency of 65% is obtained; for the antibody, it is 74%; whereas for the green fluorescent protein coding plasmids, a very small transfection efficiency is gained. If the transfection efficiency is improved, gold nanoparticles will be very useful as mediator for gene transfection in living cells.