1987 Fiscal Year Final Research Report Summary
Study on the improvement of research method of immunoelectron microscopy
| Project/Area Number |
61870072
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| Research Category |
Grant-in-Aid for Developmental Scientific Research
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| Allocation Type | Single-year Grants |
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| Research Field |
Morphological basic dentistry
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| Research Institution | Kyushu University |
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Principal Investigator |
KOJIRO KURISU Kyushu University, Faculty of Dentistry, Professor, 歯学部, 教授 (50028346)
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| Co-Investigator(Kenkyū-buntansha) |
KENGO NAGATA Kyushuu University, Faculty of Dentistry, Assistant, 歯学部, 助手 (90189134)
YASUYOSHI OHSAKI Kyushuu unuveristy, Faculty of Dentistry, Assistant, 歯学部, 助手 (70117076)
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| Project Period (FY) |
1986 – 1987
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| Keywords | Immunohistochemistyr / Electron microscopy / Fixation / Embeddig / Collagen / Protein A-fold / Post-embedding / フラルダイト |
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| Research Abstract |
In this project, we examined to appropriate condition of (1) fixative,(2) embedding materials and (3) pretreatment of ultrathin sections, for immunoelectron microscopic localization of heat-sensitive proteins. Results obtained were as follows.
1) Condition of fixatives: 2-0.1% glutaraidehyde (GA) plus 2)4% paraformaldehyde (PA) in 0.1M cacodylate buffer of PLR fixative were examined for the prefixative. for the post-fixative, 0.1% osmium treraoxide in 0.1M phoshate buffrr used always. 0.1% %g%a plus 2% PA gave result in the presearcation of both ultrastructure and antigenicity.
2) Kind of embedding materials; Epon 812 and Araidite both of which were epoxy resin were examined. Although tge preservation of ultrastracture was slightly superor in Epon than in Araldite, the preservation of antigenicity was greatly superior in Araldite than in Epon.
3) Pretreatment of the ultrathin sections: Among several agents examined acidified 10% eroxide gave base result in immunostaining intersity. %it is thought taht acidefied peroxide oxidized and solubilized reduced osmium and thus bleaches the embedded antigens.
4) Immunoelectron microscopic localization of type i and type iii collagen on the ultrathin sections pretrated with acidified peroxide of tissues prefixed with 0.1% GA plus 2% PA and embedded in Araldite gave facorable results.
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