Local variations in specific membrane lipids are thought to influence the activity of membrane proteins.  Recent application of imaging mass spectrometers and isotopic labeling techniques to cells has allowed lipid species to be detected at a resolution in parity with their organization in the cell membrane.  The development of membrane protein probes that can be simultaneously imaged with the isotopic labels for lipids would allow definitive measurements of the extent of lipid-protein colocalization.  For this purpose, we have synthesized fluorine-functionalized colloidal gold immunolables that enable imaging specific proteins in parallel with lipids in the plasma membrane with high resolution SIMS performed on a NanoSIMS.  The fluorine in the monolayer enables NanoSIMS detection, whereas the antibody allows for the selective labeling of specific proteins.  With the combination of these immunolabels and the metabolic labeling of lipids, specific membrane proteins and cellular lipids can be imaged parallel using NanoSIMS.  This method can be extended to other proteins and lipids and provide high sensitivity and lateral resolution, to investigate other reports of colocalization.

Preparation of functionalized immunolabels that permit simultaneous imaging of specific proteins and lipids in membranes with high-resolution secondary ion mass spectrometry

243rd National Meeting (2012)

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