Gliding motility assay using 16551mM 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 16551mM 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分析实验。 • 实验所用缓冲液为PEM（又称BRB80），体系中添加了16551mM重水（D₂O）。 • 图像采集使用搭载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电子倍增电荷耦合器件（Electron Multiplying Charge-Coupled Device, EMCCD）相机完成拍摄。 • 该文件集为3组重复实验中的第1组，对应15个感兴趣区域（Region of Interest，ROI）中的第11个。 • 本次成像实验旨在探究滑行运动分析中，因体系添加重水所引发的微管滑行速度变化情况。 • 实验通过物镜加热器将载玻片温度恒定维持在32℃。观测早期采集的1-5号ROI对应的微管滑行速度较慢，这是由于载玻片在观测前于室温下制备，观测过程中温度逐渐升高；在5号ROI之后，微管滑行速度趋于稳定。 • 采用载物台测微尺标定得到像素长度为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。需提前安装Mencoder工具，具体执行指令为：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。