Birefringence imaging directly reveals architectural dynamics of filamentous actin in living growth cones
Birefringence imaging directly reveals architectural dynamics of filamentous actin in living growth cones
复制标题
DOI：
10.1091/mbc.10.1.197
复制
发表时间：
1999-01-01
影响因子：
3.3
通讯作者：
Oldenbourg, R
中科院分区：
文献类型：
--
作者：
Katoh, K;Hammar, K;Oldenbourg, R
我们用一种新型的偏光显微镜（Pol-Scope）研究了神经生长锥板状脂质体中细胞骨架精细结构的动态行为。Pol-Scope图像显示具有精致的分辨率和清晰度双折射精细结构，如细丝和膜，而无需用外源染料或荧光标记处理细胞。此外，测量的蛋白质纤维的双折射在薄lamellipodial区域可以解释在纤维束中的长丝的数量。我们证实，双折射纤维肌动蛋白为基础的使用传统的荧光标记方法。通过记录随时间推移的Pol-Scope图像的电影，我们分析了在前进的生长锥中放射状纤维、丝状伪足和伪足内的产生和动态组成。在时间流逝的Pol-Scope图像中提供的严格定量信息证实了先前推断的行为，并为丝状肌动蛋白的结构动力学提供了新的见解。
We have investigated the dynamic behavior of cytoskeletal fine structure in the lamellipodium of nerve growth cones using a new type of polarized light microscope (the Pol-Scope). Pol-Scope images display with exquisite resolution and definition birefringent fine structures, such as filaments and membranes, without having to treat the cell with exogenous dyes or fluorescent labels. Furthermore, the measured birefringence of protein fibers In the thin lamellipodial region can be interpreted in terms of the number of filaments in the bundles. We confirmed that birefringent fibers are actin-based using conventional fluorescence-labeling methods. By recording movies of time-lapsed Pol-Scope images, we analyzed the creation and dynamic composition of radial fibers, filopodia, and intrapodia in advancing growth cones. The strictly quantitative information available in time-lapsed Pol-Scope images confirms previously deduced behavior and provides new insight into the architectural dynamics of filamentous actin.