CYTOPLASMIC LOADING OF DYES, PROTEIN AND PLASMID DNA USING AN IMPACT-MEDIATED PROCEDURE

We describe a method and apparatus designed to rapidly and reproducibl y produce transient, survivable plasma membrane disruptions-''wounds'' -in order to gain access to the cytoplasm of eukaryotic cells growing in culture. Compressed gas is used to propel glass beads, dispersed as a uniform aerosol, at adherent cells growing on a culture substratum. The impact of beads with the cells creates plasma membrane wounds. Ma cromolecules, such as dyes, proteins and plasmid DNAs, diffuse from th e extracellular environment directly into the cytoplasmic compartment of the cell through these wounds. Resealing of the plasma membrane, ne cessary for cell survival, traps macromolecules within the cytoplasm o f the cell.