SDS-digested freeze-fracture replica labeling(SDS-FRL) was innovated as a suitable technique to reveal the fine structure of plasma membrane and quantitative localization of biomolecules over the membrane structure. However, it was difficult to apply this

SDS-FRL是一种创新的技术，可用于揭示质膜的精细结构和在膜结构上定量定位生物分子。然而，该技术难以定量地应用于固定在原生质面（p面）上的分子。因此，在本研究项目中，我尝试优化SDS-FRL的标记方案。在SDS处理、预处理和阻断复制膜以及各标记溶液上的组分等步骤进行优化。因此，尽管不同实验的标记方案细节不同，但新方案不仅对外质面分子提供了高灵敏度的标记，而且对p面分子也提供了高灵敏度的标记。这些调查获得的定位数据作为原始论文发表。

SDS-digested freeze-fracture replica labeling(SDS-FRL) was innovated as a suitable technique to reveal the fine structure of plasma membrane and quantitative localization of biomolecules over the membrane structure. However, it was difficult to apply this technique to molecules immobilized on protoplasmic faces(P-face) quantitatively. Thus, in this research project, I tried to optimize labeling protocols of SDS-FRL. The optimization was carried out at steps of SDS treatment, pretreatment and blocking of replicated membranes as well as components on each labeling solutions. As a result, although details of labeling protocol differ among experiments, new protocol provided high-sensitive labeling not only for molecules on exoplasmic faces but also for those on P-faces. Localization data obtained by these investigations were published as original papers.