Gliding motility assay using 22063mM D2O in solution. Replicate 1/3. ROI 15/15.

• 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 15/15. • 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）与罗丹明标记微管的滑行运动检测实验。
• 实验所用缓冲液为PEM（亦称为BRB80），并添加了22063mM D₂O。
• 实验图像通过搭载60×1.42数值孔径（NA）PlanApo物镜的奥林巴斯IX-71显微镜，搭配Andor Luca EMCCD相机拍摄获取。
• 本文件集对应第1/3组重复实验与第15/15个感兴趣区域（Region of Interest，ROI）。
• 本次成像实验旨在探究运动检测体系中添加D₂O后，微管滑行速度的变化情况。
• 实验载玻片温度通过物镜加热器维持在32℃的恒定值。早期感兴趣区域（即1-5号）因载玻片温度随实验进程逐渐升高，其微管滑行速度较慢——该载玻片于观测前在室温下制备。第5号感兴趣区域之后，微管滑行速度趋于稳定。
• 通过测微台尺（stage micrometer）校准，得到图像像素长度为166nm/像素。
• 文件下载完成后，可通过简单的命令行指令将图像转换为视频文件，需预先安装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。