Gliding motility assay using 16551mM D2O in solution. Replicate 1/3. ROI 8/15.

• 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 8/15.• 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.• 本实验为基于驱动蛋白-1（kinesin-1）与罗丹明标记微管的滑行运动分析实验。 • 所用缓冲液为PEM（又称BRB80），并添加了16551mM重水（D₂O）。 • 图像采集使用搭载Andor Luca电子倍增型电荷耦合器件（Electron Multiplying Charge-Coupled Device, EMCCD）相机的奥林巴斯IX-71型显微镜，搭配60×1.42NA PlanApo物镜完成拍摄。 • 本文件集对应第1/3组重复实验与第8/15号感兴趣区域（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后，微管滑行速度的变化情况。 • 通过物镜加热器将载玻片温度恒定维持在32℃。早期感兴趣区域（即1-5号）因载玻片在观测前于室温下制备，温度随实验进程逐渐升高，导致滑行速度偏慢；5号区域之后滑行速度趋于稳定。 • 利用载物台测微尺（Stage Micrometer）校准得到像素长度为166nm/像素。 • 数据集下载完成后，可通过简单的命令行指令将图像转换为视频。需预先安装Mencoder软件，执行命令为：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（Application Programming Interface, API）完成上传。