Delivery of biomolecules into targeted single cells in living vertebrate embryos

Manipulation of gene function in vivo is an indispensable technique for modifying individual functions of targeted cells in living animals. Recently, femtosecond laser has been used for delivery of exogenous genes into cultured cell and this technique is called photoporation. However, it was difficult to achieve the single-cell modification for living animals by photoporation utilizing femtosecond laser oscillator with a low pulse energy and a high reception rate. For the first time, we applied femtosecond laser amplifier with a high pulse energy and a low repetition rate to the photoporation and realized to deliver several kinds of biomolecules, such as dextran, morpholino oligonucleotides, and DNA plasmids, into targeted single cells of zebrafish, chick, shark, and mouse embryos.

<Figure 1>

<Figure 1>Femtosecond laser was focused on Zebrafish embryo mounted in methylcellulose solution including fluorescent bio-molecules. Fluorescence was observed in three single cells (arrowheads), near which the femtosecond laser was irradiated.

Yoichiroh Hosokawa, Haruki Ochi, Takanori Iino, Akihiro Hiraoka, Mikiko Tanaka, "Photoporation of biomolecules into single cells in living vertebrate embryos induced by a femtosecond laser amplifier ," PLoS ONE Vol. 6, No. 11, pp. e27677_1- e27677_7. (2011)