FEMTOSECOND-LASER SETUPS FOR CELL-MEMBRANE PORATION

摘要

Focused femtosecond-laser pulses can create tiny transient holes in cell membranes which temporarily allows foreign genetic material from the outside to reach the cell interior. With suitable laser parameters this all optical "optoporation" allows highly efficient laser-assisted cell transfection by reprogramming the cell's genetic code with very high cell survival rates. Furthermore, the use of viruses or nanoparticle as carriers which may cause serious side effects can be completely omitted. However, the cell positions need to be precisely determined to allow individual focusing of the laser radiation onto the cell membranes which is for large cell numbers quite elaborate and time consuming. We addressed these issues and present optical microscope add-ons for fast and almost hands-off laser-assisted poration of cell membranes with automated determination of the positions of adherent cells in a culture dish or targeting continuously flowing cells in suspension.