| Approach . | How it works . | Advantages . | Caveats . |
|---|---|---|---|
| Fluorescent lipids | -Fluorescent lipids added to cells -Trafficking assessed by microscopy | -Monitor trafficking in real time in live cells | -Fluorescent groups can alter physical properties and metabolism of lipids -Fluorescent lipids may affect cells |
| Labeling & fractionation | -Labeled lipid precursors added to cells -Cells fractionated | -Allows simultaneous analysis of transport and metabolism -Direct measurement of lipid levels in organelles | -Fractionation challenging -Analysis requires cell destruction |
| Lipid modification | -Lipid modifying enzyme localized in compartment outside synthesis site -Labeled lipid precursors added to cells | -Does not require fractionation | -Lipid modification must be faster than transport -Modified lipids may affect cells -Requires effective localization of modifying enzyme -Analysis requires cell destruction |
| Lipid sensors | Fluorescent or luminescent sensors indicate changes in membrane lipid composition | -Monitor trafficking in real time in live cells -Can detect small changes in membrane lipid composition | -Factors other than lipid concentration may affect sensor binding -Sensors may perturb membranes |
| Approach . | How it works . | Advantages . | Caveats . |
|---|---|---|---|
| Fluorescent lipids | -Fluorescent lipids added to cells -Trafficking assessed by microscopy | -Monitor trafficking in real time in live cells | -Fluorescent groups can alter physical properties and metabolism of lipids -Fluorescent lipids may affect cells |
| Labeling & fractionation | -Labeled lipid precursors added to cells -Cells fractionated | -Allows simultaneous analysis of transport and metabolism -Direct measurement of lipid levels in organelles | -Fractionation challenging -Analysis requires cell destruction |
| Lipid modification | -Lipid modifying enzyme localized in compartment outside synthesis site -Labeled lipid precursors added to cells | -Does not require fractionation | -Lipid modification must be faster than transport -Modified lipids may affect cells -Requires effective localization of modifying enzyme -Analysis requires cell destruction |
| Lipid sensors | Fluorescent or luminescent sensors indicate changes in membrane lipid composition | -Monitor trafficking in real time in live cells -Can detect small changes in membrane lipid composition | -Factors other than lipid concentration may affect sensor binding -Sensors may perturb membranes |