Gliding motility assay using 16551mM D2O in solution. Replicate 3/3. ROI 11/15.

• 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 3/3 and ROI 11/15. • 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.

• 本数据集为采用驱动蛋白-1（kinesin-1）与罗丹明标记微管开展的滑行运动测定实验。
• 实验所用缓冲液为PEM（又称BRB80），其中添加了16551mM 重水（D₂O）。
• 图像采集使用搭载Andor Luca EMCCD相机的Olympus IX-71型显微镜，搭配60× 1.42NA PlanApo物镜完成。
• 该文件集对应第3组重复实验（全组共3组）及第11个感兴趣区域（ROI, Region of Interest，全区域共15个）。
• 本图像采集实验旨在探究滑行运动测定体系中，因添加重水所导致的微管运动速度变化。
• 通过物镜加热器将载玻片温度恒定维持在32℃。早期感兴趣区域（即第1至5组）的滑行速度较慢，原因在于载玻片于观测前在室温下制备，后续温度逐渐升高；第5组感兴趣区域之后，滑行速度趋于稳定。
• 使用载物台测微尺校准后，得到像素长度为166nm/像素。
• 文件下载完成后，可通过一条简单的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 API完成上传。