Gliding motility assay using 22063mM D2O in solution. Replicate 3/3. ROI 5/15.

• 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 3/3 and ROI 5/15. • 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测定实验。 • 实验所用缓冲液为PEM（又称BRB80），其中添加了22063mM重水（D₂O）。 • 图像采集使用奥林巴斯（Olympus）IX-71显微镜，搭配安道尔（Andor）Luca EMCCD相机，采用60×1.42NA PlanApo物镜。 • 本文件集为第3组重复实验（共3组），以及第5个感兴趣区域（Region of Interest, ROI，共15个）。 • 本次成像实验旨在探究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℃。前期采集的1-5号感兴趣区域，因载玻片在观测前于室温下制备，温度随实验逐渐升高，导致滑行速度偏慢；5号ROI之后，滑行速度趋于稳定。 • 使用台尺（stage micrometer）标定得到像素长度为166nm/像素。 • 文件下载完成后，可通过简单命令行指令将图像转换为视频，需预先安装Mencoder。具体指令为：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。