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 fungusUstilago maydisby 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.

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