A Method to Visualize the Actin and Microtubule Cytoskeleton by Indirect Immunofluorescence

The cytoskeleton provides the basic architectural organization and shape of the eukaryotic cell, and plays a key role in segregation of the genetic material. A method to visualize the actin and microtubule cytoskeleton in the fungus Ustilago maydis by indirect immunofluorescence is described here. The method entails growth of cells to early logarithmic phase, fixation with a cross-linking agent or organic solvent, partial digestion of the cell wall and permeabilization of cells with a detergent to allow entry of antibodies, exposure to primary antibody, followed by treatment with secondary antibody conjugated to a fluorophore to allow visualization with fluorescence microscopy.