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

• 本实验为基于驱动蛋白-1（kinesin-1）与罗丹明（rhodamine）标记微管的滑行运动测定实验。
• 所用缓冲液为PEM（又称BRB80），并添加了22063mM重水（D₂O）。
• 图像采集使用奥林巴斯（Olympus）IX-71显微镜，搭载安多（Andor）Luca电子倍增电荷耦合器件（Electron Multiplying Charge-Coupled Device，EMCCD）相机，搭配60×、1.42数值孔径（NA）的PlanApo物镜完成。
• 该文件集对应第1/3组重复实验与第11/15个感兴趣区域（Region of Interest，ROI）。
• 本次图像采集旨在探究滑行运动测定实验中，重水添加对微管滑行速度的影响。
• 载物台温度通过物镜加热器恒定维持在32℃。早期感兴趣区域（即1至5号）因观测前载物台于室温制备，实验过程中载物台温度逐渐升高，导致微管滑行速度偏慢；5号区域之后，滑行速度趋于稳定。
• 通过载物台测微尺（stage micrometer）校准，像素长度为166nm/像素。
• 文件下载完成后，可通过简单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。