Functional Analysis of Muscle Specimen by X-ray Cryomicrodiffraction Technique

X 射线冷冻微衍射技术对肌肉标本的功能分析

• 批准号: 15500294
• 负责人: IWAMOTO Hiroyuki
• 金额: $ 2.43万
• 依托单位: Japan Synchrotron Radiation Research Institute
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15500294/
• 关键词: myofibril; X-ray microdiffraction; X-ray microbeam; quick-freezing; pinhole optics; synchrotron radiation; high-flux X-ray beam; equatorial reflections; X線回折; 昆虫飛翔筋; 脊椎動物骨格筋; 単離心筋細胞; 微小領域

The system for recording microdiffraction patterns from micrometer-sized quick-frozen hydrated biological specimens at the high-flux beamline BL40XU of SPring-8 is described. The optics consists of a pair of pinholes drilled on tantalum substratum, with a defining aperture of 2 μm in diameter. The frozen specimens are placed in an in-vacuum cryochamber mounted on a three-axis goniometer, where the specimens are stably held at a liquid-nitrogen temperature (〜-199℃). A beam size of 1.5 μm (full width at half maximum) is attained at the sample position. By using this system, diffraction patterns have been recorded from an isolated single myofibril (diameter, 〜3 μm) of an insect flight muscle in an area equivalent to a single sarcomere (length, 〜3 μm). The technique is potentially applicable to other micrometer-sized hydrated biological specimens, which are more susceptible to radiation damage than dry synthetic polymers or biopolymers. The quick-freezing of biological specimens has also been proven useful in reducing the specimen volume in the beam in conventional diffraction recordings.

本文介绍了在SPring-8的高通量光束线BL 40 XU上记录微米级速冻水合生物样品的显微衍射图样的系统。光学元件由一对在钽基底上钻的小孔组成，小孔直径为2 μm。将冷冻标本置于安装在三轴测角仪上的真空冷冻室中，标本稳定地保持在液氮温度（100 - 199 ℃）下。在样品位置获得1.5 μm的光束尺寸（半高宽）。利用该系统，在相当于一个肌节（长约1.3 μ m）的面积上，记录了昆虫飞行肌的一根肌原纤维（直径约1.3 μm）的衍射图。该技术可能适用于其他微米级的水合生物标本，这是更容易受到辐射损伤比干燥的合成聚合物或生物聚合物。生物样本的快速冷冻也被证明是有用的，在传统的衍射记录中减少光束中的样本体积。

项目成果

Phylogeny of winged insects viewed from flight muscle myofilament lattice structure : An X-ray cryomicrodiffraction study.

从飞行肌肌丝晶格结构观察有翅昆虫的系统发育：X射线冷冻微衍射研究。

• 发表时间: 2005
• 期刊: Biophys.J. 88・1
• 作者: Iwamoto;H.;Wakayama;J.;Tamura;T.;Yagi;N.
• 通讯作者: N.

X-ray microdiffraction and conventional diffraction from frozen-hydrated biological specimens.

冷冻水合生物样本的 X 射线微衍射和常规衍射。

• 发表时间: 2005
• 期刊: J.Synchrotron Rad. 12
• 作者: Iwamoto;H.;Inoue;K.;Fujisawa;T.;Yagi;N.
• 通讯作者: N.

X-ray cryomicrodiffraction of myofibrils

肌原纤维的 X 射线冷冻微衍射

• 发表时间: 2004
• 期刊: Biophys.J. 86・1
• 作者: Iwamoto;H.;Wakayama;J.;Tamura;T.;Yagi;N.
• 通讯作者: N.

Hiroyuki Iwamoto, Jun'ichi Wakayama, Takumi Tamura, Naoto Yagi: "X-ray cryomicrodiffraction of myofibrils"Biophysical Journal. 86(1). 567a (2004)

Hiroyuki Iwamoto、Junichi Wakayama、Takumi Tamura、Naoto Yagi：“肌原纤维的 X 射线冷冻微衍射”生物物理学杂志。