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2015 年度 実施状況報告書

単細胞・多細胞生物進化の熱力学的解明

研究課題

研究課題/領域番号 15K13617
研究機関埼玉大学

研究代表者

中林 誠一郎  埼玉大学, 理工学研究科, 教授 (70180346)

研究期間 (年度) 2015-04-01 – 2017-03-31
キーワードC-dot / fluorescent probes / bioimaging
研究実績の概要

Highly luminescent ultrasmall carbon nanodots (CDs) have been prepared by one step microwave-assisted pyrolysis and functionalized with fluorescein photosensitizer by diazo-bond. The absorption edge of such prepared Fluorescein;CDs was red shifted in comparison with the bare one. Nevertheless, the emission signal induced by the nanoparticle quantum-sized graphite structure was quenched due to photo-isomerization of the diazo group at photoexcited state. In order to restrict the photoisomerization, i.e. rotation around nitrogen; nitrogen bond, the diazo group was fixed by metal cation to form complex compound or chelate. The obtained metal;complex of Fluorescein;N=N;CDs show absorbance maximum same as bare CDs, but recovered emission signal from nanoparticle moiety, which was bathochromic ; shifted. They exhibit lower quantum yield in comparison with the bare CDs but better photostability toward emission quenching in nutrition cell culture. The formed photosensitizer-conjugates nanoprobes were proposed as multifunctional fluorophores for intracellular in vivo imaging due to their attractive photophysical attributes, tunable and excitation-dependent emission. The bio-application of photosensitizer;conjugated CDs was demonstrated as fluorescent tracers for endocytosis pathways in cultured Tobacco cells. Their successful staining and lower toxicity to the plant cells were compared with conventional quantum dots (CdSe/ZnS core ; shell type, which caused acute toxicological in vivo effect).

現在までの達成度 (区分)
現在までの達成度 (区分)

2: おおむね順調に進展している

理由

We have reported new findings as follows;
(1).Carbon nano-dots (CDs) were conjugated with dye molecule (Fluorescein).(2).The coupling between CDs and dye was through diazo bond.(3).When the N=N- rotation was prohibited by Ruthenium chelation, new electronic state was emerged between CDs and Fluorescein. [Fluorescein;CDs]Ru2 (4). [Fluorescein;CDs]Ru2 is used as bio-imaging marker in BY2 Tobacco plant cells.(5).The carbonbased nontoxic fluorescent nanoprobes was successful in non-breaching staining with tunable visible emission color.

今後の研究の推進方策

BY-2 cells stained with Fluorescein;CDs possessed significantly stronger emission signal than those stained with [Fluorescein;CDs]Ru2+ . This effect was explained with the different quantum yield of the nanoprobes , which effect their emission signal in the fluorescence microscope. In addition, in the staining with [Fluorescein;CDs]Ru2+ we observed some labeled details surrounding cell walls, which do not appear in the staining with Fluorescein;;CDs. We also noticed that after longer incubation, a strong fluorescence was observed at the nucleus. In addition, non homogeneous staining of dividing cells was discovered in the case of staining with Fluorescein;CDs. The investigation of these phenomena is still in progress All these features demonstrate that the photosensitizer-conjugated CDs have good biocompatibility and make them appropriate candidates for cell imaging.

次年度使用額が生じた理由

193円の余りは、消耗品の端数である。

次年度使用額の使用計画

消耗品に使用する。

  • 研究成果

    (2件)

すべて 2016

すべて 雑誌論文 (2件) (うち国際共著 1件、 査読あり 2件)

  • [雑誌論文] Photosensitizer-Conjugated Ultrasmall Carbon Nanodots as Multifunctional Fluorescent Probes for Bioimaging2016

    • 著者名/発表者名
      A.Loukanov, R.Sekiya, M.Yoshikawa, N.Kobayashi,Y. Moriyasu, S. Nakabayashi
    • 雑誌名

      J.Phys.Chem.C

      巻: 120 ページ: 15867-15874

    • DOI

      10.1021/acs.jpcc.5b11721

    • 査読あり / 国際共著
  • [雑誌論文] Quantitative Evaluation of Cancer Cell Adhesion to Self-Assembled Monolayer-Patterned Substrates by Reflection Interference Contrast Microscopy2016

    • 著者名/発表者名
      T.Matsuzaki, I. Kosaku, K. Masuda, E. Kakinuma, R. Sakamoto, K. Iketaki, H.Yamamoto, M. Suganuma, N. Kobayashi, S. Nakabayashi, T. Tanii, HY.Yoshikawa.
    • 雑誌名

      J.Phys.Chem.B.

      巻: 120 ページ: 1221-1227

    • DOI

      10.1021/acs.jpcb.5b11870

    • 査読あり

URL: 

公開日: 2017-01-06  

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