Preparative methods for imaging plasmodesmata at super-resolution.

Much of our knowledge of plasmodesmata has come from the ability to visualize them. Light microscopy is a popular tool for exploring subcellular structures but is limited in its resolving power due to the diffractive properties of light. At 50 nm in diameter plasmodesmata are below this limit and so cannot be resolved. Super-resolution microscopy operates beyond the limits of conventional light microscopy affording a more detailed view. Although lacking the ultrastructural resolving power of the electron microscope (EM), super-resolution microscopy helps to bridge the gap between conventional light microscopy and EM.Here we present three preparative methods for studying plasmodesmata at super-resolution using 3D-structured illumination microscopy (3D-SIM).