Development of microscope system for single fluorescent molecule imaging in vivo

单分子成像技术在日本发展以来，已被广泛应用于分子马达的体外分析和质膜信号转导的单分子测量等研究领域。然而，这种方法是基于全内反射显微镜，这限制了其应用到距离盖玻片表面小于100 nm的区域。在这个项目中，我们试图打破这一限制，观察细胞深处的单个分子。通过将Nipow型共聚焦显微镜和高灵敏度低噪声相机相结合，我们成功地观察了细胞内部单个分子的动力学。我们进一步将该技术应用于细胞内部的超分辨率成像。

Since the development of the single molecule imaging technique in Japan, it has been applied to wide areas of researches including in vitro analysis of molecular motors and single molecule measurements of signal transduction at the plasma membrane. However, this method is based on the total internal reflection microscopy, which limited its application to the region closer than 100 nm from the cover glass surface. In this project, we have tried to break this limitation and to observe single molecule deep inside the cell. By the combination of a Nipow-type confocal microscope and a high sensitivity-low noise camera, we have successfully observe the dynamics of single molecule deep inside the cell. We have further applied this technique for the super resolution imaging deep inside the cell.