A novel microsurgery method for intact plant tissue at the single cell level using ArF excimer laser microprojection

Shin'ichiro Kajiyama, Takeshi Shoji, Shinya Okuda, Yoshihiro Izumi, Ei Ichiro Fukusaki, Akio Kobayashi

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

A novel microsurgery technique for the partial removal of rigid cell-walls in intact plant tissue is established. Using a size-variable slit, an ArF excimer laser was microprojected on the surface of the targeted cell, and this method enabled the area- and depth-controllable processing of the cortical structure of plant cells including the cuticle and cell wall layer. In epidermal cells of all tested plants, viabilities of more than 90% were retained 24 h after irradiation. Scanning electron microscope (SEM) observation revealed that the cuticle layer of the irradiated region was completely ablated, and the cellulose microfibrils of the secondary cell wall were partially removed; furthermore, 4 days after laser treatment, the regeneration of cell wall fibrils was observed. As a model experiment, the transient expression of synthetic green fluorescent protein (sGFP) was performed by the microinjection of cauliflower mosaic virus (CMV) 35S promoter-derived sGFP gene through an "aperture" in the treated cell surface. Moreover, micron-sized fluorescent beads were successfully introduced by the same method into the onion cells indicating that this method can be used to introduce foreign materials as large as organelles.

Original languageEnglish
Pages (from-to)325-331
Number of pages7
JournalBiotechnology and Bioengineering
Volume93
Issue number2
DOIs
Publication statusPublished - Feb 5 2006
Externally publishedYes

Fingerprint

Excimer Lasers
Microsurgery
Excimer lasers
Cell Wall
Cells
Tissue
Green Fluorescent Proteins
Caulimovirus
Microfibrils
Proteins
Onions
Plant Cells
Microinjections
Viruses
Cellulose
Organelles
Regeneration
Lasers
Electron microscopes
Genes

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

Cite this

A novel microsurgery method for intact plant tissue at the single cell level using ArF excimer laser microprojection. / Kajiyama, Shin'ichiro; Shoji, Takeshi; Okuda, Shinya; Izumi, Yoshihiro; Fukusaki, Ei Ichiro; Kobayashi, Akio.

In: Biotechnology and Bioengineering, Vol. 93, No. 2, 05.02.2006, p. 325-331.

Research output: Contribution to journalArticle

Kajiyama, Shin'ichiro ; Shoji, Takeshi ; Okuda, Shinya ; Izumi, Yoshihiro ; Fukusaki, Ei Ichiro ; Kobayashi, Akio. / A novel microsurgery method for intact plant tissue at the single cell level using ArF excimer laser microprojection. In: Biotechnology and Bioengineering. 2006 ; Vol. 93, No. 2. pp. 325-331.
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