Establishment of embedment-free electron microscopy and analysis of the nature of cytoplasmic matrix by this methodology

免包埋电子显微镜的建立及利用该方法分析细胞质基质的性质

基本信息

  • 批准号:
    62480092
  • 负责人:
  • 金额:
    $ 4.29万
  • 依托单位:
  • 依托单位国家:
    日本
  • 项目类别:
    Grant-in-Aid for General Scientific Research (B)
  • 财政年份:
    1987
  • 资助国家:
    日本
  • 起止时间:
    1987 至 1988
  • 项目状态:
    已结题

项目摘要

Using polyethylene glycol (PEG), a highly water soluble wax as a transient embedding media, embedment-free sections of chemically fixed tissues are obtained reliably for the electron microscopic examination. After critical-point drying with CO_2, these embedment-free sections present the same aspect of cell ultrastructure as whole mounted, fixed, and dried cultured cells, and filamentous elements (microtrabeculae), which have been unrecognized before in the conventional cytoplasmic matrix, are clearly revealed. Wehen the embedment-free sections are rotaly-replicated with platinum and carbon, resulting images of the sections are of high quality, with excellent resolution and to be quite comparable to those obtained with the rapid freezing, deep etched replica method.With this new methodology, the membrane specializations on the outer and inner cell surfaces as well as the organization of the cytoskeleton is clearly demonstrated. However, after the attainment of the intracellular displacement of cell organelles and some proteins by ultracentrifugation, no marked change in organization of the microtrabecular lattice is discerned.This together with my previous finding that artificial protein solutions at certain concentration exhibit meshworks quite similar to the intracellular microtrabeculae,argue against the idea that gthe filamentous or midrotrabecular strands are the real structure of living cells. Regerdless of the reality or artifact of the microtrabeculae, the PEG method is also applicable to general scanning electron microsoopy, and the intermicroscopic correlation of images between scanning and transmission electron microscopy and light microscopy is easily and reliably performed. Furthermore, the PEG-method is shown to be suitable for light and electron microscopic immunocytochemistry. Therefore this methodology should prove to be valuable adjunct to conventional microscopic techniques.
使用聚乙二醇(PEG),一种高度水溶性的蜡作为暂时包埋介质,化学固定的组织的无包埋切片可靠地获得用于电子显微镜检查。在临界点CO_2干燥后,这些无包埋切片呈现出与整个固定、固定和干燥的培养细胞相同的细胞超微结构,并清晰地显示出以前在常规细胞质基质中未被识别的丝状成分(微梁)。韦亨将无包埋切片用铂和碳旋转复制,所得切片图像质量高,分辨率好,与快速冷冻、深蚀刻复制法相当,这种新方法清楚地显示了细胞内外表面的膜特化以及细胞骨架的组织。然而,在细胞器和某些蛋白质通过超离心达到细胞内置换后,微小梁晶格的组织没有明显的变化,这与我以前的发现一起,在一定浓度下的人工蛋白质溶液显示出与细胞内微小梁非常相似的网状结构,反驳了丝状或中小梁链是活细胞真实的结构的观点。PEG法不考虑微小梁的真实性或伪影,也适用于普通扫描电镜,扫描和透射电镜与光学显微镜之间的显微图像相关性容易且可靠。此外,PEG-方法被证明是适用于光和电子显微镜免疫细胞化学。因此,这种方法应被证明是有价值的辅助传统的显微镜技术。

项目成果

期刊论文数量(40)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Correlative Microscopy in Biology:Instrumentation and Methods. Academic Press Inc. 347-354 (1987)
生物学中的相关显微镜:仪器和方法。
  • DOI:
  • 发表时间:
  • 期刊:
  • 影响因子:
    0
  • 作者:
  • 通讯作者:
近藤尚武: 新潟医学会雑誌. 101. 361-366 (1987)
近藤直武:新泻医学会杂志 101. 361-366 (1987)
  • DOI:
  • 发表时间:
  • 期刊:
  • 影响因子:
    0
  • 作者:
  • 通讯作者:
H.Kondo: Exp.Brain Res.Ser.16. 139-144 (1987)
H.Kondo:Exp.Brain Res.Ser.16。
  • DOI:
  • 发表时间:
  • 期刊:
  • 影响因子:
    0
  • 作者:
  • 通讯作者:
近藤尚武: Experimental Brain Research Series 16 Springer-Verlag Berlin Heidelberg. 16. 139-144 (1987)
Naotake Kondo:实验脑研究系列 16 Springer-Verlag Berlin Heidelberg。16. 139-144 (1987)
  • DOI:
  • 发表时间:
  • 期刊:
  • 影响因子:
    0
  • 作者:
  • 通讯作者:
H. Kondo: "The cytoplasmic matrix of the adrenal chromaffin cells of rats under normal condition and restraining stress" J. Electr. Microsc. Tech. Suppl.in press. (1989)
H. Kondo:“正常条件和抑制应激下大鼠肾上腺嗜铬细胞的细胞质基质”J. Electr。
  • DOI:
  • 发表时间:
  • 期刊:
  • 影响因子:
    0
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KONDO Hisatake其他文献

KONDO Hisatake的其他文献

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{{ truncateString('KONDO Hisatake', 18)}}的其他基金

Analysis of Novel Cellular Functions of Fatty Acid Binding Proteins
脂肪酸结合蛋白的新细胞功能分析
  • 批准号:
    18390056
  • 财政年份:
    2006
  • 资助金额:
    $ 4.29万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
Functional analysis of fatty acid binding proteins in immune and neural tissues.
免疫和神经组织中脂肪酸结合蛋白的功能分析。
  • 批准号:
    14370002
  • 财政年份:
    2002
  • 资助金额:
    $ 4.29万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
Molecular and Celluler Biological Analysis of the functional Significance of Phosphoinositide Metabolism
磷酸肌醇代谢功能意义的分子和细胞生物学分析
  • 批准号:
    11694235
  • 财政年份:
    1999
  • 资助金额:
    $ 4.29万
  • 项目类别:
    Grant-in-Aid for Scientific Research (A)
Molecular and Cell Biological Analysis of lipid kinases and phosphatases involved in phosphoinosilide signaling
参与磷酸肌醇信号转导的脂质激酶和磷酸酶的分子和细胞生物学分析
  • 批准号:
    11470001
  • 财政年份:
    1999
  • 资助金额:
    $ 4.29万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B).
The regulation mechanism of lipid kinase in the signal transduction
脂质激酶在信号转导中的调控机制
  • 批准号:
    09044248
  • 财政年份:
    1997
  • 资助金额:
    $ 4.29万
  • 项目类别:
    Grant-in-Aid for international Scientific Research
Molecular and Cell Biological Analysis of Lipid Kinases in Relation to Signaling and Vesicle Traffic.
与信号传导和囊泡运输相关的脂质激酶的分子和细胞生物学分析。
  • 批准号:
    09470001
  • 财政年份:
    1997
  • 资助金额:
    $ 4.29万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
Moleculap biological and Pustochemical study on phospholipid me tabolic enzymes inviolved in signal transduction
参与信号转导的磷脂代谢酶的分子生物学和化学研究
  • 批准号:
    07457001
  • 财政年份:
    1995
  • 资助金额:
    $ 4.29万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
Molecular biological and morphological analysis of signal transduction mechanism from membrane to nuchreos
从膜到核的信号转导机制的分子生物学和形态学分析
  • 批准号:
    07307027
  • 财政年份:
    1995
  • 资助金额:
    $ 4.29万
  • 项目类别:
    Grant-in-Aid for Scientific Research (A)
Regulation of inositol phospholipid-pelated 2nd messengers
肌醇磷脂包裹的第二信使的调节
  • 批准号:
    07044216
  • 财政年份:
    1995
  • 资助金额:
    $ 4.29万
  • 项目类别:
    Grant-in-Aid for international Scientific Research
Histological study on the gene expression of several proteins related to the intracellular Ca-signals.
与细胞内 Ca 信号相关的几种蛋白质的基因表达的组织学研究。
  • 批准号:
    04404020
  • 财政年份:
    1992
  • 资助金额:
    $ 4.29万
  • 项目类别:
    Grant-in-Aid for General Scientific Research (A)
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