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

• 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 3/15.• 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。 • 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 EMCCD相机的Olympus IX-71显微镜，搭配60× 1.42NA PlanApo物镜完成拍摄。 • 本文件集对应第2/3次重复实验与第3/15个感兴趣区域（Region of Interest，ROI 3/15）。 • 本次图像采集旨在探究运动分析实验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°C。前5个感兴趣区域（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，随实验进程温度逐渐升高；第5个感兴趣区域之后，滑行速度趋于稳定。 • 利用载物台测微尺（stage micrometer）标定得到像素长度为166nm/像素。 • 文件下载完成后，可通过以下简单命令行指令将图像转换为视频，需预先安装Mencoder软件：`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 API完成上传。