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Studies of ultra structures in 3-D on developmental mechanisms of biological crystals.

生物晶体发育机制的 3D 超微结构研究。

基本信息

批准号：
15390548
负责人：
WAKITA M.
金额：
$ 9.28万
依托单位：
Hokkaido University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2006
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-15390548/
关键词：
hardtissue crystal grouth electron microscopy 3-D structure development 結晶生成三次元構築

项目摘要

The cryo-ultramicrotomy for cell membrane experiences could not have appropriate instrumentation for the aim of this research until the end of research period. So we made preparation of specimen by embedding specimen using only metyl-metacrylate method.As mentioned on research abstract proposed last year (2005), the monomer of metyl-metacrylate we use has highly volatile so we made much newly devised methods for polymerization of this resin. At first, we made to polymerize metyl-metacrylate in the small specimen bottle with tight cap. It could do polymerization with minimum volatilization.We made further to work it by two step embedding method, that is, half part of monomer was polymerized in the bottle before embedding specimen, then the specimen was set on the polymerized resin and pours rest of monomer for thoroughly polymerization. According to this embedding method, we could have the specimen floating in the larger resin block. After polymerization, the bottle was broken to have t … More he resin block, and then the specimen was sawed out to make suitable size of specimen block. Because the block is too large, it is disadvantage we have to make longer work for getting the experimental block, and always to watch not missing the specimen direction. In this year, we tried to shape blocks with keeping the specimen direction from the lateral surfaces on the way to the final shape.We could not solve in the end of research period the problem of putting the ultra-thin sections on a slide glass for later treatment of de-embedding. We had the plan that sections put on the slide glass with the epoxy cement were rinsed by monomer to make tissue without resin. But we could find any appropriate method for it.Totally we could step on the primary research scheduled in the beginning, because we needed too much time for losing those problems. However, for those four years, each investigator of our team worked eagerly on this research, and at the same time each of us achieved good results personally as described on the report. Less

低温超微切片的细胞膜经验，直到研究期结束时才有合适的仪器进行研究。因此，我们只采用甲基丙烯酸甲酯包埋法制备样品。正如去年（2005）提出的研究摘要所提到的，我们使用的甲基丙烯酸甲酯单体具有高挥发性，因此我们设计了许多新的聚合方法。首先，我们将甲基丙烯酸甲酯聚合在密闭的小样品瓶中，这样可以在挥发最小的情况下进行聚合。我们进一步采用两步包埋法进行研究，即在包埋试样前，先将一半单体在瓶中聚合，然后将试样置于聚合树脂上，将剩余的单体倒入瓶中进行彻底聚合。根据这种包埋方法，我们可以使试样漂浮在较大的树脂块中。聚合后，将瓶破碎，得到更多的树脂块，然后将试样锯出合适尺寸的试样块。由于块体过大，在获取实验块的过程中需要较长时间的工作，并且要时刻注意不丢失试样方向。在这一年里，我们试图塑造块与保持试样的方向从侧面到最终形状的方式。在研究结束时，我们无法解决将超薄切片放在玻片上进行后期脱埋处理的问题。我们的计划是用环氧水泥把切片放在玻片上，用单体冲洗，制成不含树脂的组织。但我们可以找到任何合适的方法。我们完全可以按一开始的计划进行初级研究，因为我们需要太多的时间来丢失这些问题。然而，在这四年里，我们团队的每一位研究者都在积极地进行这项研究，同时我们每个人都取得了报告中所描述的良好的个人成果。少