Microtubule Gliding Assay [dataset]
These data are images of gliding motility assays using kinesin-1 and 29% rhodamine tagged microtubules. Each image was taken with an Andor Luca S EMCCD camera with a gain of 150 and an exposure of 100 ms. A total of 600 frames at 5 frames/second were gathered using a 100 W mercury lamp attenuated by 94% with a neutral density filter. Each subfolder with images was approximately 2 minutes total exposure. The microscope used was an Olympus IX71 microscope with a PlanApo 60x 1.42 NA objective that was temperature stabilized at 33.1 C. The pixel size was measured to be 166.7 nm/pixel and the images taken were 496 x 658 pixels for a field of view of approximately 83 x 110 microns. Each folder (alpha, beta, kappa, whole, and mixed) contain three assays where 20 different regions on a slide had images captured. The different labeled folders used 1.0 mg/mL of alpha (Sigma C6780), beta (Sigma C6905), kappa (Sigma C0406), whole (Sigma C7078) or mixed bovine casein as the surface passivator in the gliding motility assay. The "mixed casein" was an approximate reconstitution of the different constituents of whole casein in the ratio found in bovine casein (49% α, 37% β, and 14% κ). The goal of this experiment was to measure speed variations of microtubules in the gliding motility assay dependent on the type of surface passivator used.
Maloney, Andy; Koch, Steve (2011): Microtubule Gliding Assay. University of New Mexico. http://hdl.handle.net/1928/12559