IMAGING BARRIERS TO MOLECULAR DIFFUSION BY PAIR CORRELATION FUNCTIONS

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 我研究了细胞内扩散的模型基板（NLS-GFP）通过使用一种新的和非常规的荧光相关技术，对相关函数的基础上。我通过测量单个分子在细胞内从一个特定位置移动到另一个位置所需的时间来检测分子运输事件。这是相对于传统FCS方法的新奇，该方法测量分子穿过激光束所需的时间。通过这种方法，可以在细胞隔室（对自由扩散的限制）和穿过核膜屏障（由于核孔通道的存在而受到限制）两者内检测到扩散障碍。在这两种情况下，测量提供了在单分子水平和活细胞中NLS-GFP运输的动力学步骤的定量描述，而不需要复杂的标记程序和细胞操作。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. I study the intracellular diffusivity of a model substrate (NLS-GFP) by using a novel and unconventional fluorescence correlation technique, based on pair correlation functions. I detect molecular transport events by measuring the time a single molecule takes to move from a specific location to another within the cell. Here is the novelty with respect to the traditional FCS approach that measures the time a molecule takes to cross the laser beam. By this approach obstacle to diffusion can be detected both within cellular compartments (restrictions to free diffusion) and across the nuclear envelope barrier (restrictions due to the presence of nuclear pore channels). In both cases the measurement affords a quantitative description of the kinetic steps of NLS-GFP transport at the single molecule level, and in living cells, without the need for complex labeling procedures and cell manipulations.