The development of fluorescence-based molecular imaging has revolutionized cell biology allowing the visualization of specific biomolecules at the microscopic and, more recently, at the nanoscopic scale while in their relevant biological contexts. Nonetheless, despite the imaging toolkit for biologists interested in exploring the subcellular localization and dynamics of proteins and nucleic acids has expanded exponentially in the last decades, the means to visualize and track lipids in cells did not develop to the same extent until recently. Here we described some basic fluorescence-based techniques that can be used in standard cell biology laboratories to visualize subcellular pools of specific lipids and to evaluate their regional metabolism. Specifically, here we focus on the imaging-based analysis of phosphoinositide and sphingolipid metabolism at the Golgi complex.
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