Membrane surgery and nanosurgical cell isolation using high-intensity femtosecond laser pulses is reported. We demonstrate the applicability of using ultrashort (femtosecond) laser pulses for performing surgery on live mammalian cells. When sub-10 femtosecond pulses were focused onto the cell, precise sub-micron surgical cuts were made on the biological membrane. Traversing the cells relative to the focused laser spot, we report on localized nanosurgical ablation of focal adhesion adjoining epithelial cells. In each study, the surgery was conducted without morphologically compromising the cells. © 2005 Wiley-Liss, Inc.