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新型捞片器具在昆虫触角连续切片中的应用
The application of a new fishing apparatus in obtaining serial ultrathin sections of insect antennae
戎念杭1** 付丙鲜2*** 祝增荣3 洪 健1
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DOI:10.7679/j.issn.2095-1353.2017.125
作者单位:1. 浙江大学农生环测试中心,杭州 310058;2. 浙江农林大学农业与食品科学学院, 临安 311300;3. 浙江大学昆虫科学研究所,杭州 310058
中文关键词:外骨骼,感器,外周神经支配,透射电镜,狭缝载网
英文关键词: exoskeleton, sensilla, peripheral innervations, transmission electron microscope, slot grids
中文摘要:

【目的】 透射电子显微镜技术在生物学研究中发挥着越来越深入广泛的作用,通过透射电镜技术获得生物组织连续切片,从而对其神经网络等延伸结构进行连续追踪及三维重构是非常迫切的科研诉求。但捞片方法的限制使得连续切片的获得困难重重,尤其是对于组织致密的昆虫外骨骼如昆虫触角而言,常因包埋剂浸透不够充分而导致聚合欠佳,增加了连续切片收集的难度。为解决捞取昆虫触角连续切片过程中的系列问题,获得完整、平整的触角连续切片,我们自行研制了一种适用于狭缝载网捞片的组合器具。【方法】 用组合器具中的捞样器和放样台分别捞取及放置超薄切片。【结果】 捞取的昆虫触角切片不会局部脱离树脂切片而产生不同程度的折叠,不会整片脱离树脂切片而滑落至树脂切片一侧,样品切片完整无破损、无皱褶,连续切片成条贴附在狭缝覆膜区。【结论】 新型捞片器具为获得昆虫触角连续切片提供了有力支持。

英文摘要:

[Objectives]  Transmission electron microscopy is playing a more and more important role in biological research, including tracking extended structures such as neural networks and three dimensional reconstruction. Consequently, the demand for continuous sections of biological tissues for transmission electron microscopy has greatly increased. However, limitations of the current fishing method make the acquisition of continuous sections difficult, especially when samples are comprised of dense insect exoskeleton, such as continuous sections of the antennae. Insufficient penetration of the agent usually causes poor polymerization, which undoubtedly increases the difficulty of collecting intact serial sections. To solve these problems, we developed a kind of combined device for collecting sections with slot grids. [Methods]  A hybrid device comprised of a special fishing loop and a lofting platform was developed and its ability to fish for serial sections and place them onto slot grids was evaluated. [Results]  No partial detachment and folding occurred in the serial sections obtained by this combined device; specimen sections neither separated completely, nor slid to the side of the resin section. Sample sections were intact without any damage or wrinkles, and serial sections clung to the slot area in a line. [Conclusion]  The new device is an improvement on previous methods of obtaining continuous sections of insect antennae for electron microscopy. 

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