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標識金微粒子の円偏光二色性を指標とした生体内分子重合の解析

Research Project

Project/Area Number 62890004
Research Category

Grant-in-Aid for Developmental Scientific Research

Allocation TypeSingle-year Grants
Research Field 広領域
Research InstitutionNagoya University

Principal Investigator

佐藤 英美  名古屋大学, 理学部, 教授 (40109260)

Co-Investigator(Kenkyū-buntansha) 竹中 裕  株式会社ニコン, 光学部, 部長
藤原 敬己 (藤原 敬巳)  国立循環器病センター, 循環形態部, 部長 (10190092)
御橋 広真  名古屋大学, 理学部, 助教授 (30022594)
黒田 英世  名古屋大学, 理学部, 助教授 (50064845)
石川 春律  群馬大学, 医学部, 教授 (90010058)
Project Period (FY) 1987 – 1989
Project Status Completed (Fiscal Year 1989)
Budget Amount *help
¥18,800,000 (Direct Cost: ¥18,800,000)
Fiscal Year 1989: ¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 1988: ¥6,800,000 (Direct Cost: ¥6,800,000)
Fiscal Year 1987: ¥8,400,000 (Direct Cost: ¥8,400,000)
Keywords超顕微鏡 / 金微粒子標識法 / 高検出能 / 画像記録 / 偏光光学系 / 金コロイド / 円偏光二色性 / 高解像画像記録 / 顕微注射
Research Abstract

本年度は試験研究の最終年度である。従って私達は超顕微鏡の調整と、高感度・高忠実画像記録装置を組み合せたシステム化に努力した。
超顕微鏡光源は輝度が不足している。均質な照明をえる為に敢てハロゲン光源とファイバ-グラスを用いたが、超高圧水銀灯と高屈折率の熔融石英ファイバ-を結合させることで高輝度光源がえられることが判った。超顕微鏡の検出能を高める為に、将来この改良光源も取り付けられるよう改造したい。また、60×NA1.40のアポクロマ-トレンズの質が向上したので、これにレクチファイヤ-を装着し、より優れた偏光光学系とするよう予備試験を行った。改良は着実に進行していると考える。
<in>___ー vitroの試行として、アクチン分子を重合条件でスライドグラスにのせ、アクチン繊維の形成と会合を顕微鏡下で追跡した。その結果、殆んど教本のアクチン繊維が次第に束ねられ、所謂ストレスファイバ-となる過程をVTR記録することに成功した。同様に、繊維芽細胞の弱複屈折性ストレスファイバ-の生体内における動態の記録にも成功した。直径5nmの金標識抗体の細胞内顕微注射はまだ試験段階にあり、混合比、注射量の決定、蛋白標識率の測定等をテスト中である。予備実験では、5nm金コロイド粒子は、条件をととのえれば確実に輝点としてトレ-スできるので、当初の意図はほぼ達成されたと信じている。この超顕微鏡の2号機、3号機は既にニコンエンジニアリングで受注している。
高感度、高解像画像記録についてはカタログ通りの性能がえられず、相つぐ試行錯誤の結果、浜松ホトニクスの協力をえて、どうにか及第できる所迄きた。
これ等の試行結果をまとめ、光学の可能性の限界に挑戦した記録としての報告書を作成し、近い将来に解決すべき問題点を具体的に列記して役に立つ参考資料として配布する準備を進めている。

Report

(2 results)
  • 1989 Annual Research Report
  • 1988 Annual Research Report
  • Research Products

    (40 results)

All Other

All Publications (40 results)

  • [Publications] Sato,H.,Kobayashi,A.&Itoh,J.T.: "Molecular basis of physical and chemical,probes for spindle assembly." Cell Struct.and Funct.,. 14. 1-34 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Sumitro,S.B.&Sato,H.: "The isotope effects of D_2O in developing sea urchin eggs." Cell Struct.and Funct.,. 14. 95-112 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Cachon,J.,Sato,H.,Sachon,M.&Sato,Y.: "Analysis by polarizing microscopy of chromosomal structure among dinoflagellates and its phylogenetic involvement." Biol.of the Cell.,. 65. 51-60 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Yokota,E.,Mabuchi,I.,Kobayashi,A.&Sato,H.: "Effects of Acylpeptide K-26 on the motility of sea urchin sperm model.I.Inhibitin of the motility of both live and triton X-100 rxtracted aperm." Cell Struct.and funct.,. 14. 299-310 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Yokota,E.,Mabuchi,T.,Kobayashi,A.&Sato,H.: "Effects of Acylpeptide K-26 on the motility of sea urchin sperm model. II.Mechanism of the inhibitory effect of K-26." Cell Struct.and funct.,. 14. 311-320 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Sumitoro,S.B.Izutsu,K.&Sato,H.: "Effects of D_2O on the movement of chromosomes and the shorteming of Kinetochore spindle fibers in anaphase in dividing spermatocytes of the grasshopper,Monogolotettix japonicus." Cell Strukt.and funct.,. 14. 345-352 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] 佐藤英美・伊藤知彦・小林昭雄: "核分裂抑止剤による分裂細胞中心体の構造変化の誘導。" “蛋白質・核酸・酵素". 34. 1610-1617 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] 佐藤英美・竹中裕・二星俊明: "超顕微鏡の試作。" “生物物理". 29. 42-45 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Shimada,O.&Ishikawa,H.: "Somatostatin-containing neurons in the brain:An immuno-histochemical study and comparison with the rat brain." Arch.Histol.Cytol.52. 201-212 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Katoh,K.&Ishikawa,H.: "The cytoskeletal involvement in cellularization of the Drosophila melanogaster." Protoplasma. 150. 83-95 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] 石川春律: "組織細胞の極性化と細胞骨格。" 蛋白質核酸酵素. 34. 1742-1748 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] K.Mihashi,A.Ooi,N.Suzuki,H.Yoshimura,&K.Kinosita,Jr.: "Dynamic Structure of F-actin and Its Ca-Dependent Regulation under the Influence of Tropomyosin-Troponin System." Life Science Advances,. 7. 22-28 (1988)

    • Related Report
      1989 Annual Research Report
  • [Publications] N.Suzuki,and K.Mihashi: "Subunit-Flow in F-actin under Steady-State Condition." Biophys.Chem.33. 177-193 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] K.Mihashi,N.Suzuki,and A.Ooi: "Ca-dependent Requlation of the Dynamic Polarity of F-actin under the Influence of Tropomyosin and Troponin." Biophys.Chem.33. 195-204 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] K.Mihashi,A.Ooi and T.Hiratsuka: "Evidence for the Existence of Two Equilibrium Conformations of the ternary Complex of Myosin subfragment-1,ADP and Orthovanadate." J.Biochem.(in press;due in March. 106. (1990)

    • Related Report
      1989 Annual Research Report
  • [Publications] Kuroda,H.,Kuroda,R.and Sakai,T.: "Membrane electrogenesis in plasmodia of Physarum polycephalum:A dominant role for the proton pump." Biochim.Biophys.Acta(Biomembranes). 987. 154-164 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Obata,S.and Kuroda,H.: "Membrane potential change in a sea urchin egg induced by calcium ionophore(A23187)has the same component that is inbolved in the fertilization potential." Cell Struct.Funct.14. 697-706 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Ballarin-Denti,A.,Slyman,C.L.and Kuroda,H.: "Lipid Soluble distribution probes are mismeasures of membrane potential in walled eukaryotic cells." In plant Membrane Transport:Current position (ed.,F.Rasi-Caldogno,E.Demichaelis and E.Marre)Elsevier Science Publishers,Amsterdam,. 79-80 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Ballarin-Denti,A.,Slyman,C.L.And Kuroda,H.: "Complex interactions of lipid soluble ions with wall-bearing eukaryotic cells." To be submitted to Biochim.Biophys.Acta October(in press).(1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Obata,S.and Kuroda,H.: "Voltage-independent component of the fertilization current in sea urchin eggs." Dev.Biol.(in press).

    • Related Report
      1989 Annual Research Report
  • [Publications] Onishi,H.,Maita,T.,Matsuda,G.and Fujiwara,K.: "Evidonce for the association between two myosin heads in rigor acto-smooth muscl heavy meromyosin." 1989.Biochemistry.28. 1898-1904 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Onishi,H.,Maita,T.,Matsuda,G.and Fujiwara,K.: "The carbodiimido-catalized cross-linking sites in the heads of gizzard heavy meromyosin attached Lo F-actin." 1989.Biochemistry.28. 1905-1912 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] 藤原敬己,Fitjgerald,M.L.,神宮司洋一: "高血圧と血管内皮細胞の細胞骨格" 生体の科学. 40. 202-207 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] 藤原敬己: "細胞骨格成分の細胞内分布" 細胞工学. 8. 453-460 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] 藤原敬己: "ストレスファイバ-の形態と機能" 蛋白質核酸酵素. 34. 1513-1523 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] McBeath,E.and Fujiwara,K.: "Coalignment of microtubules,cytokeratin intermediate filaments,and collagen in a collagen secreting cell system." Eur.J.Cell Biol.50. 510-521 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Masuda,M.and Fujiwara,K.: "Cultured endothelial colls respond to fluid shcar stress." In Etiology and Morphogenesis <of>___ー Congenital Heart Disease.

    • Related Report
      1989 Annual Research Report
  • [Publications] Masuda,M.and Fujiwara,K.: "Motility of cultured endothelial cells under fluid shear stress." Ann.N.Y.Acad.Sci.(1990)

    • Related Report
      1989 Annual Research Report
  • [Publications] Onishi,H.and Fujiwara,K.: "The rigor configuration of smooth muscle heavy meromyosin trapped by a zero-length cross-linker." Biochemistry.(1990)

    • Related Report
      1989 Annual Research Report
  • [Publications] McBeath,E.and Fujiwara,K.: "Microtubule detachment from the MTOC as a keyevent in the coplete turnover of microtubules in cells." Eur.J.Cell Biol.(1990)

    • Related Report
      1989 Annual Research Report
  • [Publications] 増田道隆・藤原敬己: "血流と内皮細胞の方向性" 臨床医学(印刷中). (1990)

    • Related Report
      1989 Annual Research Report
  • [Publications] 藤原敬己: "細胞接着とストレス線維" 生体の科学(印刷中). (1990)

    • Related Report
      1989 Annual Research Report
  • [Publications] 佐藤英美: 生体の科学. 38. 467-470 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] Sato,H.;Bryan,J.: Zoological Science.5. 623-638 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] Kuroda,H.;Obata,S.;Takemoto,K.;Ishiguro,M.;Sato,H.: Zoological Science.5. 733-742 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] Amaniya,S.;Uemura,I.;Shinkai,T.;Sato,H.: Exptl.Cell Res.179. 446-453 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] Kobayashi,A.;Hino,T.;Yata,S.;Ito,T.J.;Sato,H.;Kawazu,K.: Agric.Biol.Chem.52. 3119-3123 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] 佐藤英美: 病態生理. 7. 820-822 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] 佐藤英美: "細胞工学6__ー:No.3." 秀潤社, 246-249 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] 佐藤英美: "生体の科学、<39>___ー:別刷"細胞光学特性測定法"" 医学書院, 530-533 (1988)

    • Related Report
      1988 Annual Research Report

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Published: 1987-04-01   Modified: 2016-04-21  

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