Gliding motility assay using 11040mM D2O in solution. Replicate 3/3. 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 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 3/3 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.

• 本数据集为使用驱动蛋白-1（kinesin-1）与罗丹明标记微管开展的滑行运动测定实验。
• 所用缓冲液为PEM（又称BRB80），并添加了11040mM重水（D₂O）。
• 图像采集采用搭载于奥林巴斯IX-71显微镜的Andor Luca EMCCD相机，搭配60× 1.42NA PlanApo物镜完成拍摄。
• 本文件集为第3组重复实验（共3组），且为第1个感兴趣区域（Region of Interest，ROI，共15个）。
• 本次成像旨在探究滑行运动测定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°C。前5个感兴趣区域（ROI 1-5）的微管滑行速度较慢，这是由于观测前载玻片在室温下制备，随拍摄进程载玻片温度逐渐升高；ROI 5之后滑行速度趋于稳定。
• 使用台尺（stage micrometer）校准得到像素标定长度为166nm/像素。
• 文件下载完成后，可通过简单的命令行指令将图像序列转换为视频。需预先安装Mencoder工具，执行指令为：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。