Gliding motility assay using 22063mM D2O in solution. Replicate 1/3. 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 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 1/3 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.

• 本数据集对应一项使用驱动蛋白-1（kinesin-1）与罗丹明标记微管的滑行运动测定实验（gliding motility assay）。
• 实验所用缓冲液为PEM（又称BRB80），并添加了22063mM的重水（D₂O）。
• 图像采集使用搭载于奥林巴斯IX-71显微镜的Andor Luca EMCCD相机，搭配60×1.42NA PlanApo物镜完成拍摄。
• 本文件集对应第1/3组重复实验与第10/15个感兴趣区域（Region of Interest, ROI）。
• 本次图像采集旨在探究运动测定实验中添加重水所引发的微管滑行速度变化。
• 载玻片温度通过物镜加热器维持在32℃恒定值。早期感兴趣区域（即1-5号ROI）的滑行速度较慢，这是由于载玻片在观测前于室温下制备，随观测进程温度逐渐升高；而在5号ROI之后，滑行速度趋于稳定。
• 采用载物台测微尺（stage micrometer）标定后，测得像素长度为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（Application Programming Interface, API）完成上传。