Photoinjection of Fluorescent Nanoparticles into Plant Cells Using Femtosecond Laser Amplifier with Enzyme Treatment

摘要

Femtosecond laser photoinjection has become a popular method to deliver various kinds of molecules such as genes,proteins, and fluorescent dyes into single mammalian cells. However, this method is not easily applied to plant cells. Thegrowth of vacuoles in plant cell increases inner pressure (turgor pressure) which makes injection difficult through diffusionprocess. Furthermore, the plant cell membrane is protected by a thick rigid cell wall. Normally, only substances with diameter< 5 nm can pass through the cell wall. Therefore, it would be an additional barrier for large substances. This work isa demonstration of the photoinjection of large objects (fluorescent nanoparticles, diameter = 80 nm) into cytoplasm of anintact single plant cell. A femtosecond laser amplifier was employed for the photoinjection under moderate enzyme treatmentconditions.