Gliding motility assay using 11040mM 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 11040mM 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.

• 本实验为基于驱动蛋白-1（kinesin-1）与罗丹明标记微管的滑行运动分析实验。
• 所用缓冲液为PEM（又称BRB80），并添加了11040mM的重水（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物镜完成拍摄。
• 本文件集对应第1/3次重复实验及第15/15个感兴趣区域（Region of Interest，ROI）。
• 本次成像实验旨在探究滑行运动分析体系中，因添加重水所引发的微管滑行速度变化情况。
• 载玻片温度通过物镜加热器维持在32℃恒定。早期采集的感兴趣区域（ROI 1-5）对应的微管滑行速度较慢，这是由于观测前于室温制备的载玻片温度仍在逐步升高；待采集至第5个感兴趣区域后，微管滑行速度趋于稳定。
• 通过载物台测微尺（stage micrometer）校准，得到像素长度为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。需预先安装Mencoder软件，具体命令如下：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。