Plasmon-resonant nanoparticles attached to cell membranes, under laser treatment can temporarily increase membrane permeability. In this paper, the influence of continuous-wave and pulsed (nanosecond) laser irradiation on living cells incubated with gold nanoparticles was investigated. Gold nanospheres, nanostars, and nanorods with different functionalization were used as plasmonic agents. The dependence between increase of medium temperature on the irradiation time was showed for nanostars and nanorods with different surface properties. Cells samples incubated with gold nanorods showed the highest temperature increase. Feasibility of cell optoporation by the use of gold nanospheres with variable functionalization was demonstrated. The cell membrane permeability was successfully enhanced as shown by the uptake of the fluorescent dye upon nanosecond laser treatment. Toxicity of the nanoparticles was estimated by MTT assay.