Gene delivery process in a single animal cell after femtosecond laser microinjection

Yoichiroh Hosokawa*, Seriya Iguchi, Ryohei Yasukuni, Yuji Hiraki, Chisa Shukunami, Hiroshi Masuhara

*Corresponding author for this work

Research output: Contribution to journalArticle

24 Scopus citations

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.

Original languageEnglish
Pages (from-to)9880-9884
Number of pages5
JournalApplied Surface Science
Volume255
Issue number24
DOIs
StatePublished - 30 Sep 2009

Keywords

  • Cultured animal cell
  • Femtosecond laser
  • Gene expression
  • Laser microinjection

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