Gliding motility assay using 8mM D2O in solution. Replicate 2/3. ROI 4/15.

• 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 8mM 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 4/15. • 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.• 本实验为使用驱动蛋白-1（kinesin-1）与罗丹明标记微管开展的微管滑行运动测定实验。 • 实验所用缓冲液为PEM（亦称BRB80），并添加了8mM重水（D₂O）。 • 图像采集采用搭载于Olympus IX-71型显微镜上的Andor Luca型电子倍增电荷耦合器件（EMCCD）相机，搭配60× 1.42数值孔径（NA）PlanApo物镜完成。 • 该文件集对应第2/3次重复实验的第4/15个感兴趣区域（ROI, Region of Interest）。 • 本次图像采集旨在探究滑行运动实验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°C。前5个感兴趣区域（即ROI 1-5）的滑行速度较慢，这是由于载玻片在观测前于室温下制备，随观测进程推进温度逐渐升高；ROI 5之后，微管滑行速度趋于稳定。 • 使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批量转换为视频，需预先安装Mencoder工具。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, Application Programming Interface）完成上传。