Maloney, Andy Gliding motility assay using 22063mM D2O in solution. Replicate 1/3. ROI 11/15. <p>• This is a gliding motility assay using kinesin-1 and rhodamine labeled microtubules.</p> <p>• The buffer used was PEM (also known as BRB80) with the inclusion of 22063mM D2O.</p> <p>• Images were taken with an Andor Luca EMCCD camera on an Olympus IX-71 microscope using a 60x 1.42NA PlanApo objective.</p> <p>• This fileset is replicate 1/3 and ROI 11/15.</p> <p>• Images were taken to investigate speed variations of microtubules due to the inclusion of D2O in the motility assay. </p><p>• 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.</p> <p>• Pixels lengths were measured to be 166nm/pixel using a stage micrometer.</p> <p>• 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</p> <p>• Upload of these files were made possible through the figshare API.</p> microtubules;microscopy;kinesin-1;heavy water;gliding motility assay;fluorescence;Biophysics 2012-11-15
    https://figshare.com/articles/dataset/Gliding_motility_assay_using_22063mM_D2O_in_solution_Replicate_1_3_ROI_11_15_/97577
10.6084/m9.figshare.97577.v1