Gliding motility assay using 5528mM D2O in solution. Replicate 1/3. 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 5528mM 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 1/3 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.

• 本数据集为使用驱动蛋白-1（kinesin-1）与罗丹明标记微管（rhodamine labeled microtubules）开展的微管滑行运动分析实验。
• 所用缓冲液为PEM（又称BRB80），其中添加了5528mM重水（D₂O）。
• 图像采集采用搭载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 EMCCD相机完成。
• 本文件集对应3次重复实验中的第1次，涉及感兴趣区域（Region of Interest, ROI）14/15。
• 本次图像采集旨在探究该运动分析实验中添加重水对微管滑行速度的影响。
• 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，随实验进程温度逐步升高，导致微管滑行速度偏慢；5号之后的感兴趣区域滑行速度趋于稳定。
• 采用载物台测微尺校准得到像素标定长度为166nm/像素。
• 文件下载完成后，可通过以下命令行指令将图像批量转换为视频，需提前安装Mencoder软件：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（figshare API）完成上传。