Gliding motility assay using 16551mM D2O in solution. Replicate 2/3. ROI 10/15.

• 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 16551mM 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 10/15. • 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.

• 本实验采用驱动蛋白-1（kinesin-1）与罗丹明标记微管（rhodamine labeled microtubules）搭建滑行运动测定体系。
• 所用缓冲液为PEM缓冲液（又称BRB80缓冲液），并添加了16551mM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显微镜，搭配Andor Luca型电子倍增电荷耦合器件（EMCCD）相机完成。
• 本次文件集对应第2/3次重复实验与第10/15个感兴趣区域（ROI）。
• 本图像集旨在探究运动测定体系中添加重水后微管滑行速度的变化情况。
• 通过物镜加热器将载玻片温度恒定维持在32°C。早期感兴趣区域（即1-5号）因载玻片在观测前于室温下制备，后续温度升高导致滑行速度较慢；5号感兴趣区域之后，滑行速度趋于稳定。
• 利用台测微尺标定得到像素长度为166nm/像素。
• 文件下载完成后，可通过简易命令行指令将图像转换为视频。需提前安装Mencoder软件，具体指令为：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。