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

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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