Journal:

Article Title: Single Fungal Kinesin Motor Molecules Move Processively along Microtubules

doi:

Figure Lengend Snippet: Schematic illustration of the TIRF setup. The setup was built around a modified Zeiss Axiovert 135. The beam of an argon ion laser was coupled into a single mode fiber and then projected on a suprasil prism at a low, adjustable angle. The prism was optically coupled to a quartz slide with glycerol. Total internal reflection took place at the quartz-water interface. The evanescent wave of the laser beam illuminated only a very thin layer of the assay chamber allowing us to collect fluorescence near the interface with a very low background signal. Fluorescence of both the microtubule (Cy5) and the kinesin molecules in solution (Cy3) was collected with a high numerical aperture objective (Zeiss, Plan-Apochromat, 100×, NA 1.4). After locating a microtubule, Cy5 fluorescence was blocked by a 590/75LP filter set, to collect Cy3 signal only. The signal was first amplified by a Videoscope and Hamamatsu image intensifier, and the intensified image was subsequently quantified by a CCD camera. The resulting image was filtered with an image controller (Argus) before presentation on a video monitor and storage on S-VHS-tape.

Article Snippet: The signal was first amplified by a Videoscope and Hamamatsu image intensifier, and the intensified image was subsequently quantified by a CCD camera.

Techniques: Modification, Fluorescence, Amplification