The observation of growth cones with advanced optical methods

用先进的光学方法观察生长锥

• 批准号: 09680812
• 负责人: TATSUMI Hitoshi
• 金额: $ 1.92万
• 依托单位: Nagoya university
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09680812/
• 关键词: Growth Whe; evanescent; near-field; Neuron; imaging; 神経; シナプス; ビーズ; 細胞接着; V-DIC; 微分干渉顕微鏡; イメージング; グルタミン酸

Evanescent light illumination was introduced to a multi-imaging microscope to construct a new type of total internal reflection fluorescence microscope (TIRFM). The microscope combined TIRFM, high resolution video-enhanced differential interference contrast (DIC), epifluorescence, interference reflection (IR) with a state-of-the-art image acquisition system for time-lapse microscopy. Dynamic changes of the cell-substrate contact of the neuronal growth cone of a rat hippocampal neuron were studied with this TIRFM system. Growth cones were stained with membrane labeling fluorescence dyes (DiI or octadecyl rohdamine B) and observed using the multi-imaging capacities of this system. When growth cone regions were stimulated by pressure ejection of a high potassium solution, TIRF intensity at the basal membrane of the growth cone increased, suggesting that basal membrane of growth cone approaches the glass substrate when excited. The approach of the ventral membrane to the substrate during excitatory stimulation was also observed in IR mode. The functional importance of cell/substrate contact in growth cones is discussed.

将倏逝光照明引入多成像显微镜，构建了一种新型全内反射荧光显微镜。该显微镜结合了TIRFM、高分辨率视频增强微分干涉对比（DIC）、表观荧光、干涉反射（IR）和最先进的延时显微镜图像采集系统。用该系统研究了大鼠海马神经元生长锥细胞-基质接触的动态变化。用膜标记荧光染料（DiI或十八烷基罗丹明B）对生长锥进行染色，并利用该系统的多成像能力进行观察。当高钾溶液压力喷射刺激生长锥区域时，生长锥基膜处的TIRF强度增加，表明生长锥基膜在激发时接近玻璃基板。在红外模式下也观察到兴奋刺激时腹侧膜向基底的接近。讨论了生长锥中细胞/底物接触的功能重要性。

项目成果

辰巳仁史: "バイオイメージング(分担)" 共立出版, 340 (1998)

辰巳仁：“生物成像（分享）”Kyoritsu Shuppan，340（1998）

Tatsumi,H.and Y.Katayama.: "Growth cones exhibit enhanced cell-cell adhesion after neurotransmitter release." Neuroscience. accepted. (1999)

Tatsumi,H. 和 Y.Katayama.：“生长锥在神经递质释放后表现出增强的细胞间粘附力。”

辰巳仁史: "光ピンセットによる微小生物試料の操作" 応用物理. 66. 970-973 (1997)

Hitoshi Tatsumi：“使用光镊操作显微生物样品”应用物理学 66. 970-973 (1997)。

Tatsumi, H.and Y.Katayama.: "Growth cones exhibit enhanced cell-cell adhesion after neurotransmitter release" Neuroscince. (accepted). (1999)

Tatsumi, H. 和 Y.Katayama.：“生长锥在神经递质释放后表现出增强的细胞间粘附”神经科学。

Soeda, H.H, Tatsumi.and Y, Katayama.: "Neurotransrnitter release from growth cones of rat dorsal root ganglion neurons in culture." Neuroscience. 77. 1187-1199 (1997)

Soeda, H.H, Tatsumi. 和 Y, Katayama.：“培养物中大鼠背根神经节神经元的生长锥释放神经递质。”