Gliding motility assay using 11040mM D2O in solution. Replicate 2/3. ROI 7/15.

• This is a gliding motility assay using kinesin-1 and rhodamine labeled microtubules. • The buffer used was PEM (also known as BRB80) with the inclusion of 11040mM D2O. • Images were taken with an Andor Luca EMCCD camera on an Olympus IX-71 microscope using a 60x 1.42NA PlanApo objective. • This fileset is replicate 2/3 and ROI 7/15. • Images were taken to investigate speed variations of microtubules due to the inclusion of D2O in the motility assay. • The temperature of the slide was held constant at 32°C by an objective heater. Early ROIs, i.e. 1-5, have slower giding speeds due to the increasing temperature of the slide which, was prepared at room temperature before observation. After ROI 5 the gliding speed stabilizes. • Pixels lengths were measured to be 166nm/pixel using a stage micrometer. • After the files have downloaded, a simple command line entry will convert the images to a movie. Mencoder must be installed. ls > list.txt | mencoder mf://@list.txt -mf w=496:h=658:fps=30:type=png -ovc lavc -lavcopts vcodec=mpeg4:mbd=2:trell -o output.avi • Upload of these files were made possible through the figshare API.

• 本实验为采用驱动蛋白-1（kinesin-1）与罗丹明标记微管的滑行运动测定试验。
• 所用缓冲液为PEM缓冲液（又称BRB80缓冲液），其中添加了11040mM重水（D2O）。
• 图像采集使用搭载60倍1.42数值孔径PlanApo物镜的奥林巴斯IX-71型显微镜，并搭配Andor Luca型电子倍增电荷耦合器件（EMCCD）相机完成。
• 本文件集对应第2/3次重复实验及第7/15个感兴趣区域（Region of Interest, ROI）。
• 本次图像采集旨在探究滑行运动测定体系中添加重水后，微管运动速度的变化情况。
• 通过物镜加热器将载玻片温度恒定维持在32°C。前5个感兴趣区域（ROI 1-5）因观测前载玻片于室温制备、后续温度逐步升高，其微管滑行速度较慢；在第5个感兴趣区域之后，滑行速度趋于稳定。
• 采用载物台测微尺标定得到像素长度为166nm/像素。
• 文件下载完成后，可通过简单的命令行操作将图像转换为视频。需预先安装Mencoder，执行命令如下：ls > list.txt | mencoder mf://@list.txt -mf w=496:h=658:fps=30:type=png -ovc lavc -lavcopts vcodec=mpeg4:mbd=2:trell -o output.avi
• 本数据集文件通过figshare应用程序编程接口（API）完成上传。