Gliding motility assay using 8mM D2O in solution. Replicate 2/3. ROI 4/15.
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https://figshare.com/articles/dataset/Gliding_motility_assay_using_8mM_D2O_in_solution_Replicate_2_3_ROI_4_15_/97065
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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 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.
• 本数据集为采用驱动蛋白-1(kinesin-1)及罗丹明标记微管开展的滑行运动测定实验。
• 所用缓冲液为PEM(又称BRB80),并添加了8mM重水(D₂O)。
• 图像采集使用搭载60×1.42数值孔径(Numerical Aperture, NA)PlanApo物镜的奥林巴斯(Olympus)IX-71显微镜,搭配Andor Luca EMCCD相机完成。
• 本文件集为第2/3次重复实验样本,对应感兴趣区域(Region of Interest, ROI)4/15。
• 本次图像采集旨在探究运动测定实验中添加重水(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,其温度随时间逐渐升高;ROI 5之后的滑行速度趋于稳定。
• 使用载物台测微尺标定后,得到像素长度为166nm/像素。
• 文件下载完毕后,可通过简易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。示例指令如下:`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。
创建时间:
2012-11-01



