Gliding motility assay using 22063mM 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 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 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.• 本实验为采用驱动蛋白-1（kinesin-1）与罗丹明标记微管的滑行运动检测试验。 • 实验所用缓冲液为PEM（又称BRB80），添加了22063mM重水（D₂O）。 • 图像采集使用奥林巴斯IX-71型显微镜搭载Andor Luca EMCCD相机，搭配60×、1.42数值孔径的PlanApo物镜。 • 本次文件集对应第2/3组重复实验，以及第4/15个感兴趣区域（Region of Interest，ROI）。 • 本次图像采集旨在探究运动检测实验中添加重水（D₂O）对微管滑行速度的影响。 • 载玻片温度通过物镜加热器维持在32℃恒定不变。前5个感兴趣区域（ROI 1-5）的滑行速度较慢，这是因为载玻片在观测前于室温下制备，其温度随实验进程逐渐升高；在ROI 5之后，滑行速度趋于稳定。 • 通过载物台测微尺（stage micrometer）测得像素长度为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。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（Application Programming Interface，API）完成上传。