Gliding motility assay using 22063mM D2O in solution. Replicate 3/3. ROI 14/15.

• 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 22063mM 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 3/3 and ROI 14/15. • 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检测实验。
• 所用缓冲液为PEM（又称BRB80），其中添加了22063mM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，搭配安道尔Luca型电子倍增电荷耦合器件（EMCCD）相机完成。
• 本文件集为第3组重复实验（共3组），对应第14个感兴趣区域（ROI，共15个）。
• 本图像集旨在探究滑行运动检测实验中，因添加重水所引发的微管运动速度变化情况。
• 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）因载玻片在观测前以室温制备、后续温度逐步升高，导致滑行速度较慢；5号ROI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，可通过简单的命令行指令将图像转换为视频。需提前安装Mencoder，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。