Gliding motility assay using 16551mM D2O in solution. Replicate 1/3. ROI 10/15.
收藏资源简介:
• 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 1/3 and ROI 10/15. • 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.
• 本数据集为一项使用驱动蛋白-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(亦称BRB80),体系中添加了16551mM的重水(D₂O)。 • 图像采集基于奥林巴斯(Olympus)IX-71型显微镜,搭配60×1.42NA PlanApo物镜,并使用Andor Luca电子倍增电荷耦合器件(Electron Multiplying Charge Coupled Device, EMCCD)相机完成拍摄。 • 本文件集对应3组重复实验中的第1组重复,且为15个感兴趣区域(Region of Interest, ROI)中的第10个区域。 • 本次图像采集旨在探究滑行运动分析实验中因添加重水(D₂O)所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。早期的1-5号感兴趣区域滑行速度较慢,这是由于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所致;在第5号感兴趣区域之后,微管滑行速度趋于稳定。 • 通过台测微尺(stage micrometer)校准,测得图像像素长度为166nm/像素。 • 文件下载完成后,可通过一条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` • 本数据集文件可通过figshare应用程序编程接口(Application Programming Interface, API)完成上传。




