Gliding motility assay using 5528mM D2O in solution. Replicate 1/3. ROI 1/15.

• 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 1/15. • 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（rhodamine）标记的微管开展的滑行运动分析实验。
• 实验所用缓冲液为PEM（亦称BRB80），并添加了5528mM重水（D₂O）。
• 图像采集采用奥林巴斯IX-71型显微镜搭配Andor Luca EMCCD相机，使用60×1.42NA PlanApo物镜完成拍摄。
• 本文件集为第1/3组重复实验样本，以及第1/15个感兴趣区域（Region of Interest，ROI）。
• 本次成像实验旨在探究运动分析体系中添加重水对微管滑行速度的影响。
• 通过物镜加热器将载玻片温度恒定维持在32℃。前5个感兴趣区域（ROI 1-5）的滑行速度较慢，这是由于载玻片于观测前在室温下制备，导致其温度逐渐升高；ROI 5之后，微管滑行速度趋于稳定。
• 使用台尺（stage micrometer）校准得到像素标定系数为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软件，具体转换指令为：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。