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），并添加了5528mM重水（D₂O）。
• 图像采集使用搭载于奥林巴斯IX-71显微镜上的Andor Luca电子倍增电荷耦合器件（EMCCD）相机，搭配60×1.42NA PlanApo物镜完成拍摄。
• 该文件集对应3组重复实验中的第1组，以及14/15号感兴趣区域（Region of Interest，ROI）。
• 本次成像实验旨在探究滑行运动测定体系中，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。早期采集的感兴趣区域（即1-5号）因载玻片从室温升温过程，滑行速度偏慢；5号区域之后，微管滑行速度趋于稳定。
• 使用载物台测微尺（stage micrometer）校准得到图像像素尺寸为166nm/像素。
• 文件下载完成后，可通过简单的命令行指令将图像序列转换为视频，需预先安装Mencoder工具，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 API完成上传。