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- W1004801025 abstract "This chapter examines microscopic assays of axonemal dynein function and illustrates novel observations derived from these assays. Two general types of functional motility assays—namely, (1) video microscopic analysis of ATP-induced microtubule sliding in protease-treated, isolated axonemes and (2) in vitro microtubule translocation assay are described each of which makes use of dark-field light microscopy, to directly observe active microtubule sliding in isolated axonemes. The chapter also describes microscopic setup, step-by-step procedures for each experimental approach, and data derived from these techniques. The key elements of dark-field microscopy include an intense light source, a dark-field oil immersion condenser with high numerical aperture, a high-quality 40x objective lens, and a sensitive video camera mounted on a trinocular head. An upright microscope is used for experiments, which require perfusion of samples in perfusion chambers. The inverted microscope is used for most other experiments. The sample is contained in a simple perfusion chamber constructed from a glass slide and coverslip separated by two parallel strips of double-sided tape." @default.
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- W1004801025 date "1993-01-01" @default.
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- W1004801025 title "Chapter 6 Assays of Axonemal Dynein-Driven Motility" @default.
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- W1004801025 doi "https://doi.org/10.1016/s0091-679x(08)60163-6" @default.
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