Detection of single microtubules in living cells: particle transport can occur in both directions along the same microtubule.
Detection of single microtubules in living cells: particle transport can occur in both directions along the same microtubule.
复制标题
DOI：
10.1083/jcb.99.5.1785
复制
发表时间：
1984-11
期刊：
影响因子：
--
通讯作者：
Allen RD
中科院分区：
文献类型：
--
作者：
Hayden JH;Allen RD
视频增强对比度/差分干涉对比度显微镜与整体电子显微镜相结合用于研究成纤维细胞中颗粒沿线状元素的传输。取淡色林蛙角膜基质中的角质形成细胞，在金色指示器格子上培养，并用视频显微镜观察。拍摄线状元件和相关颗粒传输的视频记录，直到细胞的裂解和/或固定完成。然后对制备的材料进行整体安装的电子显微镜检查。通过结合这两种方法，我们证明了在活细胞中检测到的线状元件可以被鉴定为单个微管，而在裂解和固定细胞中可以检测到小至10 nm的细丝。溶解后，不同细胞质结构的可见度发生了变化，可见更多的细胞成分。裂解后的微管变得更难检测，而微丝束变得更加突出。观察到所有颗粒的移位都发生在由一个或多个微管组成的线状元件上。此外，还观察到颗粒在同一微管上单向或双向移动。
Video-enhanced contrast/differential interference-contrast microscopy was used in conjunction with whole mount electron microscopy to study particle transport along linear elements in fibroblasts. Keratocytes from the corneal stroma of Rana pipiens were grown on gold indicator grids and examined with video microscopy. Video records were taken of the linear elements and associated particle transport until lysis and/or fixation of the cells was completed. The preparations were then processed for whole mount electron microscopy. By combining these two methods, we demonstrated that linear elements detected in the living cell could be identified as single microtubules, and that filaments as small as 10 nm could be detected in lysed and fixed cells. The visibility of different cytoplasmic structures changed after lysis with many more cellular components becoming visible. Microtubules became more difficult to detect after lysis while bundles of microfilaments became more prominent. All particle translocations were observed to take place along linear elements composed of one or more microtubules. Furthermore, particles were observed to translocate in one or both directions on the same microtubule.