Takagi Takuya, Kii Hiroaki, Kinjo Masataka
Laboratory of Supramolecular Biophysics, Research Institute for Electronic Science, Hokkaido University, Sapporo 060-0812, Japan.
Curr Pharm Biotechnol. 2004 Apr;5(2):199-204. doi: 10.2174/1389201043376904.
FCS and FCCS measurements provide two important analytical parameters, the average number of molecules in the detection area and the translational diffusion constant of the molecules at the single molecule level. Considering these properties, FCS and FCCS have been applied to analysis of the cellular environment and dynamic processes of molecules in the living cell. More recently, a systematic approach for the analysis of macromolecule complex formation has focused on the new field of single molecule detection in the post-genome era. In this work, we tested the sensitivities of FCS and FCCS based on the distance between fluorophores in DNA as a model macromolecule complex. The results show that FCCS is not limited by the size of the macromolecular complex even in a very small detection area.
荧光相关光谱(FCS)和荧光交叉相关光谱(FCCS)测量提供了两个重要的分析参数,即检测区域内分子的平均数量以及单分子水平下分子的平动扩散常数。考虑到这些特性,FCS和FCCS已被应用于分析活细胞中的细胞环境和分子的动态过程。最近,一种用于分析大分子复合物形成的系统方法聚焦于后基因组时代单分子检测的新领域。在这项工作中,我们以DNA中荧光团之间的距离作为模型大分子复合物,测试了FCS和FCCS的灵敏度。结果表明,即使在非常小的检测区域,FCCS也不受大分子复合物大小的限制。