Demonstration of Calcium Ion Distribution in Calcifying Cells : X-ray Microanalysis and Electron Spectroscopic Imaging after Fixation with NHA-Containing Fixative and Microwave Irradiation
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概要
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We have seen reporting on the calcium ion distribution in cells and tissues with X-ray microanalysis (EDX) since 1978 [9] and with electron energy-loss spectroscopy (ESS) imaging since 1983 [10, 17], under various fixing conditions [12-16, 18-24]. Almost ten years ago, we tried a fixative containing NHA (N,N-naphthaloylhydroxamine) as a chemical precipitant for Ca<2+> ions under microwaved conditions [12, 19]. However, the data did not seem to be as sensitive or useful, under conventional electron microscopy and computerized X-ray microanalysis, compared to the two-step replacement method combined with microwave fixation using potassium (K) oxalate followed by potassium (K) antimonate, for the detection of Ca<2+> ions [12, 19-21]. Recently, however, we used ESS to analyze same tissue sections which were fixed with NHAcontaining fixative with microwave fixation ten years previously. We discovered with ESS analysis that NHA is a sensitive and ideal chemical reagent for calcium ion deteclion in a cell giving the same view as that of frozen dried or freeze-substituted tissue sections. Calcium was clearly seen and widely distributed in developing chondroblast endoplasmic ultrastructures: ribosomes, cell and endoplasmic membranes, mitochondria, Golgi, and nucleus. However, free mesen-chymal cells which were not involved in calcifying functions had only a very small amount of calcium. Calcium was widely distributed in the lacunae of the surrounding space of the chondroblasts and inmature matrix, but the exoplasm of the chondroblasts was almost calcium-free. Other elemental images in the young chondrocytes or osteoblasts, i.e.. P・L, S・L, O・K, and N・K, were seen clearly superimposed on the cell ultrastructures, i.e.. the cell membranes, ribosomes, endoplasmic reticula, mitochondria, Golgi, and nucleus [24].
- 日本組織細胞化学会の論文
著者
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Nagai Noriyuki
Department of Oral Pathology and Medicine, Graduate School of Medicine, Dentistry & Pharmaceutical S
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Mizuhira Vinci
東京医科歯科大学難治疾患研究所
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Mizuhira V
Medical Research Institute Tokyo Medical And Dental University
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Mizuhira Vinci
Department Of Anatomy And Cell Biology Medical Research Institute Tokyo Medical And Dental Universit
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AKAGI Takumi
Department of Oral Pathology, School, of Dentistry, Okayama University
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HASEGAWA Hiroshi
Research and Development Center, Toshiba Corporation
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Nagai Noriyuki
Department Of Oral Pathology School Of Dentistry University Of Okayama
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Nagai Noriyuki
Department Of Oral Pathology And Medicine Dental School Okayama University
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Akagi Takumi
Department Of Oral Pathology School Of Dentistry University Of Okayama
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Mizuhira Vinci
Department Of Anatomy & Cell Biology Tokyo Medical & Dental University
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Hasegawa Hiroshi
Research And Development Center Toshiba Corp.
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Hasegawa Hiroshi
Research Institute For New Drags Shionogi Co. Ltd.
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Akagi Takumi
Department Of Oral Pathology Okayama University Dental School
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