Abstract
This chapter focuses on the growing demand for microtubule (MT) motility assays in light of the widespread recognition of the pivotal role of microtubule (MT) motors in the cell cycle. Unstained MTs are visualized most conveniently by video microscopy using computer-enhanced differential interference contrast (DIC). MTs assembled from fluorescently labeled tubulin can be visualized using epifluorescence. DIC optics is optimized for visible wavelengths, and it is conventional to use a green interference filter to give narrow band illumination. For maximum image quality in all contrast modes, it is advisable to reduce the number of optical components between the objective and the camera to a minimum. Vibration will degrade the highly magnified image. Low frequencies will cause the image to bounce around, while high frequencies will be averaged out by the video framing rate and cause the image to blur. The effective resolution following recording can be visualized by recording and replaying a test card image having black and white lines at various spatial frequencies. Currently, the best option is digital recording to video tape. Digital video tape recorders input and output an analog signal, but encode data digitally to tape.
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