Analysis of cell poration by femtosecond laser for particle insertion by optical manipulation

摘要

The introduction and subsequent expression of external DNA inside single living mammalian cell (transfection) can be achieved by photoporation with femtosecond laser. After photoporation, external DNA can be introduced by trapping and successive insertion of DNA coated nanoparticle in the cell using optical tweezers. To maximize the transfection efficiency, one of the major aspects is that the photoporated cell should not be damaged and cell membrane should heal itself immediately or after sometime while the cells are healed in the COsub2/sub incubator. Furthermore, the size of hole created as a result of photoporation should be more than the size of DNA coated nanoparticle to be inserted inside the cell. In this paper, an analysis has been done on single cell of important breast cancer cell lines named MCF-7 and MDAMB231. Size of holes created in cell membrane after photoporation has been measured and the required optimum energy with sustained cell life were determined. Using this analysis, most favorable conditions for maximum transfection efficiency can be determined.© (2012) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.