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

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. The growth of vacuoles in plant cell increases inner pressure (turgor pressure) which makes injection difficult through diffusion process. 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 [1, 2]. This work is a demonstration of the photoinjection of large objects (fluorescent nanoparticles, diameter = 80 nm) into cytoplasm of an intact single plant cell. A femtosecond laser amplifier was employed for the photoinjection under moderate enzyme treatment conditions.