DESIGN & INSTALLATION OF AN ATTACHMENT FOR GFP PHOTOBLEACHING

（由NIH/NCRR/P41 RR 01219基金和NIH GMS 40198支持） 梭里德尔）。 表达GFP标记的融合蛋白的常规能力 细胞中的蛋白质有望成为一种重要的工具， 了解许多新蛋白质的功能， 各种基因组计划。 今年我们改进了我们的低光照水平 荧光成像系统，使得其可用于进行GFP（和 其它荧光孔）光漂白研究。 为此，我们购买了一个 廉价的单相多线氩激光器，并修改了 这样激光束就可以直接穿过 射野，然后通过物镜聚焦到样本上。 因此，我们现在可以创建一个1-2（m直径的圆形光漂白 在含有GFP或其他合适的荧光团的任何细胞中的模式。 这种增强提供了测量运动的能力， GFP分子及其周转，使用标准荧光 光漂白后恢复（FRAP）方法。 这种改进是由内部需要有选择地 光漂白中心体相关GFP标记的γ微管蛋白。

(Supported by the NIH/NCRR/P41 RR01219 grant and by NIH GMS 40198 to C. Rieder). The routine ability to expresses GFP-tagged fusion proteins in cells promises to become an important tool for understanding the function of many new proteins identified by the various genome projects. This year we improved our low light level fluorescence imaging system so that it can be used to conduct GFP (and other fluoropore) photobleaching studies. To do this we purchased an inexpensive Uniphase multi-line Argon laser and modified the microscope so that the beam of this laser could be directed through the epi-port and then focused onto the specimen through the objective. As a result we can now create a 1-2 (m diameter circular photobleached pattern in any cell containing GFP or other suitable fluorophores. This enhancement provides the capacity to measure the motility of GFP-molecules, and their turnover, using standard fluorescence recovery after photobleaching (FRAP) methods. This im provement was driven by the in-house need to selectively photobleach centrosome-associated GFP-labeled gamma tubulin.