The mammalian membrane consists of many different lipids. It is hypothesized that microdomains also called ‘rafts’ form within the membrane. These rafts are a heterogeneity in the membrane that consist of sphingolipids and cholesterol. To determine if these rafts exist the sphingolipids should be non-random orderd within the membrane.
To determine the localization of sphingolipids in the membrane several techniques can be utilized. When good probes are used to directly or via a reporter molecule label the sphingolipids several techniques can be used to visualize the localization of the sphingolipids. These techniques can be light microscopy and electron microscopy. Fluorescent probes can be used in light microscopy, but the resolution of is to low to observe lipid rafts. So in order to obtain higher resolution information Fluorescent Resonance Energy Transfer can be used. FRET can be measured between 2 different probes (hetero-FRET) and between probes of the same kind (homo-FRET). Light microscopy can easily done in living cells but it doesn’t give higher structural information so next to LM also Electron microscopy can be used. When electron microscopy is used cells need to be frozen fast in order to represent the in vivo situation.
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