Daniela Cimini, Lisa A Cameron, Edward D Salmon (2011) CIL:12019, Potorous tridactylus, epithelial cell. CIL. Dataset

Mitotic PtK1 cell microinjected with X-rhodamine-labeled tubulin and Alexa 488-labeled CENP-F antibodies to fluorescently label kinetochore fibers (red) and kinetochores/spindle poles (green), respectively. The movie starts in metaphase and shows sister kinetochore pairs aligned at the metaphase and oscillating between the two spindle poles as their respective kinetochore fibers lengthen and shorten. As the cell enters anaphase and the sister chromatids separate, sister kinetochores move away from each other as their kinetochore fibers shorten (anaphase A). Almost simultaneously, the two spindle poles move away from each other leading to elongation of the mitotic spindle (anaphase B). The compacted DNA of the chromosome arms excludes the background fluorescence so that chromosome arms appear as dark rod-like structures and the movement of the sister chromatid arms away from each other during anaphase can also be followed.

Digital images were collected with an Orca ER cooled CCD camera (Hamamatsu Photonics) coupled to a Yokogawa spinning disk confocal CS10 unit (Perkin Elmer Life Sciences Wallac), which was attached to a Nikon TE300 inverted microscope. Illumination was achieved via an Argon/Krypton laser and 488 nm and 568 nm filters in an excitation filter wheel (Sutter Instruments) through a 60 x 1.4 NA Plan-Apochromatic DIC objective lens. Nearly simultaneous fluorescence images were acquired at 488 and 568 nm at a single focal plane every 60 seconds. The movie is played at 10 frames per second. Elapsed time is shown in min:sec.

本数据集收录了经过X-罗丹明标记的微管和Alexa 488标记的CENP-F抗体荧光标记的着丝粒纤维（红色）以及着丝粒/纺锤体极（绿色）的Mitotic PtK1细胞显微注射图像。视频从有丝分裂中期开始，展示了姐妹着丝粒对在中期对齐，并在各自的着丝粒纤维伸长和缩短的过程中在两个纺锤体极之间振荡。随着细胞进入后期，姐妹染色单体分离，姐妹着丝粒在着丝粒纤维缩短的同时相互远离（后期A）。几乎同时，两个纺锤体极相互远离，导致有丝分裂纺锤体的延长（后期B）。染色体臂压缩的DNA排除了背景荧光，使得染色体臂呈现出暗色的棒状结构，并且可以追踪姐妹染色单体臂在后期相互远离的运动。数字图像采用Orca ER冷却CCD相机（Hamamatsu Photonics）采集，该相机与Yokogawa旋转盘共聚焦CS10单元（Perkin Elmer Life Sciences Wallac）相连，该单元安装在Nikon TE300倒置显微镜上。照明通过Ar/Kr激光以及激发滤光轮（Sutter Instruments）中的488 nm和568 nm滤光片实现，滤光轮通过60 x 1.4 NA Plan-Apochromatic DIC物镜。在单焦平面以每秒60秒的频率几乎同时获取了488 nm和568 nm的荧光图像。视频以每秒10帧的速度播放。显示的时间格式为分钟:秒。