Gliding motility assay using 16551mM D2O in solution. Replicate 2/3. ROI 2/15.

• 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 2/3 and ROI 2/15. • 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.• 本实验为采用驱动蛋白-1与罗丹明标记微管的滑行运动测定实验。 • 所用缓冲液为PEM（又称BRB80），并添加了16551mM D₂O。 • 实验图像通过搭载60×1.42数值孔径PlanApo物镜的奥林巴斯IX-71显微镜，搭配Andor Luca型电子倍增电荷耦合器件（Electron Multiplying Charge-Coupled Device, EMCCD）相机采集得到。 • 该文件集对应重复实验2/3，感兴趣区域（Region of Interest, ROI）2/15。 • 本次图像采集旨在探究滑行运动测定实验中，因添加D₂O而引发的微管滑行速度变化情况。 • 载玻片温度通过物镜加热器维持在32℃恒定值。早期感兴趣区域（即编号1-5）因载玻片于观测前在室温下制备，后续温度逐步升高，导致其微管滑行速度较慢；在观测至第5个感兴趣区域后，滑行速度趋于稳定。 • 使用载物台测微尺标定得像素长度为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。需预先安装Mencoder软件，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应用程序编程接口（Application Programming Interface, API）完成上传。