摘要：

This chapter provides some recipes suitable for labeling the cytoskeleton, giving examples where GFP tags and immunofluorescence can be usefully combined. Advances in live cell imaging, combined with green fluorescent protein (GFP) tags and analogs now provide independent methods for assessing the localization of molecules. Immunofluorescence microscopy is now a standard procedure for the localization of molecules in cells. In addition to facilitating the visualization of protein dynamics in vivo with sensitive imaging systems, these tagging methods can be usefully combined with the immunofluorescence procedure. One needs to drain any excess solution from the cell side by touching the edge of the coverslip to the filter paper. Then one should incubate on blocking solution for 15 rain or until first antibody mixtures are prepared. Different antibodies commonly require different fixation protocols to give optimal labeling. It is therefore important to test different fixation conditions for each antibody to determine the best compromise fixation for multiple labeling.

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