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An Improved Microscope for Bead and Surface-Based Motility Assays

  • Nick Carter
  • Rob Cross
Part of the Methods in Molecular Biology™ book series (MIMB, volume 164)

Abstract

We describe the construction and operation of an optimized microscope for kinesin motility assays, using either microtubules (MTs) sliding over kinesincoated surfaces or the motility of kinesin-coated beads on immobilized single microtubules. A Zeiss Axiovert was modified by replacing the standard Zeiss condenser and mechanical stage with an improved arrangement that provides much better mechanical stability and very high resolution differential interference contrast (DIC) imaging of unstained microtubules using standard tungsten lamp illumination. The microscope can be used routinely for MT surface-sliding assays, using low-volume flow cells constructed by sandwiching together two cover slips. The design allows for the optional addition of optical trapping and high-resolution tracking hardware, for recording of singlemolecule mechanical transients.

Keywords

Objective Lens Flow Cell Differential Interference Contrast Optical Trap Optical Trapping 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Humana Press Inc. 2001

Authors and Affiliations

  • Nick Carter
    • 1
  • Rob Cross
    • 1
  1. 1.Molecular Motors GroupThe Marie Curie Research InstituteSurreyUK

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