Tracking Movements of the Microtubule Motors Kinesin and Dynein Using Total Internal Reflection Fluorescence Microscopy.

Total internal reflection fluorescence microscopy (TIRFM) is a wide-field illumination technique that illuminates only the molecules near the glass coverslip. It has become widely used in biological imaging because it has a significantly reduced background and high temporal resolution capability. The principles of TIRFM are illustrated in this protocol, in which the movements of motor proteins are imaged as they move along microtubules within live axonemes.