Microdomains enriched with cholesterol and sphingolipids, which are referred to as lipid rafts, are believed to be present in the plasma membrane and mediate important cellular functions.   Functional tests, biochemical assays, biophysical measurements, and fluorescence microscopy have provided valuable information about cell membrane organization.  Yet, no consensus exists on the sizes and locations of compositionally distinct domains in the plasma membrane.  A direct method to image specific lipid molecules is required to unambiguously establish the distributions of cholesterol and sphingolipids in the plasma membrane.  To this aim, we have used high-resolution secondary ion mass spectrometry to chemically image the distributions of metabolically incorporated 15N-sphingolipids and 18O-cholesterol in the plasma membranes of intact fibroblast cells with approximately 90 nm lateral resolution.  We show that lipids within the plasma membrane are spatially organized on length scales that range from two hundred nanometers to a few micrometers.

High-resolution secondary ion mass spectrometry reveals the cholesterol and lipid distribution within the plasma membranes of fibroblast cells

243rd National Meeting (2012)

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