생쥐상악치조부에서의 파골세포주름변연의 당단복소재검색에 관한 전자현미경적 연구
An Electron Microscopic Study on the Detection of the Glycoprotein Localization in the Ruffled Border of the Osteoclast of the Mice Maxillary Alveolar Bone
김명국, 김철휘, 백식석,
소속 상세정보
김명국 ( ) - 서울대학교
김철휘 ( ) - 서울대학교
백식석 ( ) - 서울대학교
KMID : 0355819930170010007
Abstract
The purpose of this study was to investigate the detection of the glycoprotein localization in the ruffled border of the osteoclast of the maxilary alveolar bone in the mice. For this study male mice weighing 17gm were used.
Animals were killed by intracardiac perfusion of 30ml of 2% glutaraldehyde in 0.1M cacodylate buffer, pH 7.4. The maxillae were then removed and further fixed in Karnovsky fixative for an additional 3-4 hour. After rinsing in 0.1M cacodylate
buffer
for
10 minutes, the maxillae were demineralized for 2 weeks at 4℃ in ethylene diamine tetraacetate containing 2% glutaraldehyde. The first interdental areas were mesiodistally sectioned into slices of 1mm thickness and postfixed in osmium tetroxide.
Tissues were then dehydrated and embedded in Poly Bed. Ultra-thin sections(70~80mm in thickness) were stained with uranyl acetate and lead citrate, and examined in the electron microscope.
The interdental area of the demineralized maxilla was washed in several changes of 0.1M cacodylate buffer for 1 to 2hour at 4℃. The washed tissues were then dehydrated for 20 min in unpolymerized 90% and for 20 min in 97% glycol methacrylate
embedding
mixture(seven pars glyocl methacrylate, three parts butylmethacrylate, 2% 2, 4-dichlorobenzol peroxide paste and 3% distillled water) at 3℃. Polymerization was accomplished at 3℃ with ultraviolet light for 1 to 3 days. Ultra-thin
sections(70-80nm
in
thickness) were stained for 5 to 10 minutes by floating on a freshly prepared, filtered solution of 1% phospotungstic acid in 10% chromic acid in distilled water. After staining, the sections were floated on distilled water for a few seconds,
rapidly
transferred to formaver-coated grids, and observed with electron microscope.
Furthermore, the membrane of the ruffled border and small vesicles at the apical portion of osteoclasts revealed a strong phosphtungstic acid staining.
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