in vitro Motility Assay

Motility Assay Schematic

The in vitro motility assay involves a nitrocellulose coverslip secured to a glass slide with enough space for injecting (“flowing through”) solutions.  A saturating concentration of Myosin is flowed through the cell, adhering to the nitrocellulose, and in the presence of ATP can interact with fluorescently labeled actin filaments, propelling them across the surface.  These actin filaments are visualized with an inverted epifluorescence microscope with an attached camera and their velocities can be analyzed. ¬†Using different simulated physiological conditions found in vivo we can uncover the underlying changes to the acto-myosin interaction. 

This is an example of the videos we can create via our fluorescence camera.

 

 

 

 

 

 

 

 

 

 

 

 

 

in vitro motility assay with unregulated actin filaments at pH7.4 and 2mM ATP.