ムラマツ シュキ    MURAMATSU Shuki
   村松 朱喜
   所属
食健康科学部 健康デザイン学科
 
女性健康科学研究所 所属教員
   職種
専任講師
言語種別 英語
発行・発表の年月 2016/11
形態種別 学術雑誌
査読 査読あり
標題 A modified single-cell electroporation method for molecule delivery into a motile protest, Euglena gracilis.
執筆形態 共著
掲載誌名 Journal of Microbiological Methods
掲載区分国外
巻・号・頁 130,pp.106-111
著者・共著者 ◎Ohmachi M., Fujiwara Y., Muramatsu S., Yamada K., Iwata O., Suzuki K., and Wang O.
概要 Single-cell transfection is a powerful technique for delivering chemicals, drugs, or probes into arbitrary, specific single cells. This technique is especially important when the analysis of molecular function and cellular behavior in individual microscopic organisms such as protists requires the precise identification of the target cell, as fluorescence labeling of bulk populations makes tracking of individualmotile protists virtually impossible. Herein,we havemodified current single-cell electroporation techniques for delivering fluorescentmarkers into single Euglena gracilis, a motile photosynthetic microalga. Single-cell electroporation introduced molecules into individual living E. gracilis cells after a negative pressure was applied through a syringe connected to the micropipette to the target cell. The new method achieves high transfection efficiency and viability after electroporation. With the new technique, we successfully introduced a variety of molecules such as GFP, Alexa Fluor 488, and exciton-controlled hybridization-sensitive fluorescent oligonucleotide (ECHO) RNA probes into individual motile E.
gracilis cells. We demonstrate imaging of endogenous mRNA in living E. gracilis without interfering with their physiological functions, such as swimming or division, over an extended period of time. Thus the modified single-
cell electroporation technique is suitable for delivering versatile functional molecules into individual motile protists.