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

十二烷基硫酸钠消化的冷冻-断裂复制品标记(SDS-FRL)被创新为一种适合于揭示质膜精细结构和生物分子在膜结构上的定量定位的技术。然而，这项技术很难定量地应用于固定在原生质面(P-Face)上的分子。因此，在本研究项目中，我试图优化SDS-FRL的标记方案。优化步骤包括十二烷基硫酸钠处理、复制膜的前处理和封堵以及各标记液上的成分。因此，尽管标记方案的细节在不同的实验中有所不同，但新的方案不仅为外质面上的分子提供了高灵敏度的标记，也为P面上的分子提供了高灵敏度的标记。通过这些调查获得的本地化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.