Gliding motility assay using 11040mM D2O in solution. Replicate 2/3. 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 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 2/3 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.• 本数据集为基于驱动蛋白-1（kinesin-1）与罗丹明标记微管（microtubules）的滑行运动分析实验（gliding motility assay） • 实验所用缓冲液为PEM（亦称为BRB80），其中添加了11040mM重水（D₂O） • 成像采用奥林巴斯（Olympus）IX-71显微镜，搭配60×1.42数值孔径（NA）的PlanApo物镜，并使用Andor Luca电子倍增电荷耦合器件（EMCCD）相机采集图像 • 本文件集为第2/3次重复实验样本，对应第5/15号感兴趣区域（ROI） • 本次成像实验旨在探究运动分析实验中添加重水后微管滑行速度的变化情况 • 载玻片温度通过物镜加热器维持在32℃恒定值。前5个感兴趣区域（ROI 1-5）的滑行速度较慢，这是因为实验前在室温下制备载玻片，导致载玻片温度随实验进程逐渐升高；在ROI 5之后，滑行速度趋于稳定 • 使用载物台测微尺（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序列转换为视频，需预先安装Mencoder软件。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应用程序编程接口（API）完成上传