Gliding motility assay using 16551mM 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 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 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.

• 本实验为基于驱动蛋白-1（kinesin-1）与罗丹明（rhodamine）标记微管的滑行运动分析实验（gliding motility assay）。
• 所用缓冲液为PEM（亦称BRB80），并添加了16551mM的重水（D₂O）。
• 图像采集使用奥林巴斯（Olympus）IX-71型显微镜，搭载60×1.42NA PlanApo物镜，并搭配Andor Luca EMCCD相机完成。
• 本文件集对应第2/3次重复实验以及第4/15号感兴趣区域（Region of Interest，ROI）。
• 本次图像采集旨在探究滑行运动分析实验中添加重水（D₂O）所引发的微管运动速度变化。
• 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；而ROI 5之后，滑行速度趋于稳定。
• 通过台尺（stage micrometer）校准，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文件，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。