|
|||||
|
|
Gene delivery process in a single animal cell after femtosecond laser microinjection |
|
| Authors: | Yoichiroh Hosokawa Seriya Iguchi Yuji Hiraki Chisa Shukunami |
| Institution: | a Graduate School of Materials Science, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara, Japan b Department of Applied Physics, Osaka University, 2-1 Yamada-oka, Suita, Osaka, Japan c Institute for Frontier Medical Sciences, Kyoto University, 53 Kawahara-cho Shogoin, Sakyo-ku, Kyoto, Japan d CREST, Japan Science and Technology Agency, 4-1-8 Honcho, Kawaguchi, Saitama, Japan e Department of Applied Chemistry and Institute of Molecular Science, National Chiao Tung University, 1001 Ta-Hsueh Road, Hsinchu 30010, Taiwan |
| Abstract: | Microinjection of extracellular molecules into a single animal cell was performed by an amplified femtosecond laser irradiation. When a single-shot laser pulse was focused on the plasma membrane of a single fibroblast from the mouse cell line NIH3T3 with a high-numerical aperture objective lens, a transient hole with a diameter of 1 μm was formed. The delivery process of extracellular molecules immediately after the hole formation was monitored by a fluorescence staining with fluoresceinisothiocyanate-dextran (FITC-dextran). Then the gene expression was confirmed using a DNA plasmid of an enhanced green fluorescent protein (EGFP). The gene expression was observed when the laser pulse was focused first on the cellular membrane and then on the nuclear membrane, while the gene was not expressed when the laser was focused only on the cellular membrane. On the basis of these results, the efficiency of gene delivery by the femtosecond laser microinjection and the subsequent gene expression were clarified. |
| Keywords: | 42 62 Be (biological and medical applications) 52 38 Mf (laser ablation) 82 39 Pj (nucleic acids DNA and RNA bases) 87 17 Uv (biotechnology of cell processes) |
| 本文献已被 ScienceDirect 等数据库收录！ | |