Abstract:Fresh generative cells were isolated from mature pollen grains by means of a squash method in 7 species belonging to 3 families of angiosperms. Nomarski differential interference contrast, fluorescence, and video-enhanced microscopical studies revealed that the isolated generative cells appeared structurally intact and showed clear image of the membrane, the cytoplasm and the nucleus with 1–2 nucleoli, and the absence of a typical cell wall. It was the first time to obtain a scanning electron-microscopical image of a generative cell which became possible only after its isolation. Immunofluorescence of tubulin showed the distribution of long, mainly axial strands of the cortical microtubule. Morphologically, the isolated cells varied considerably from spindle to spherical shape, which were found to be dependent on osmolarity of the medium and treatment with the microtubule stabilizer. Fluorescein diacetate test confirmed the viability of the freshly isolated generative cells. The advantages and prospects of the isolation of generative cells are discussed.