(676h) Fluorinated Nanoparticle Immunolabels for Imaging Specific Membrane Proteins in Parallel with Cell Membrane Lipids Using High-Resolution Secondary Ion Mass Spectrometry

The cellular plasma membrane is organized into compositionally and functionally distinct domains.  Many tools have been developed to visualize protein organization within cellular membranes. Though most biological processes are executed by proteins, the local abundances of specific lipid species near a membrane protein may also influence protein activity. To characterize the relationship between protein activity and the local lipid composition, the protein of interest must be imaged in parallel with the specific lipid species.  A combination of high-resolution secondary ion mass spectrometry (SIMS) performed on a Cameca NanoSIMS and metabolic labeling enables imaging the distributions of isotope-labeled lipids in cellular plasma membranes with ~100 nm lateral resolution.  To permit imaging specific proteins in parallel with the isotope-labeled lipids using this technique, we have developed fluorine-functionalized gold nanoparticles immunolabels to tag distinct proteins.  As a proof of concept, we demonstrate imaging the distribution of influenza hemagglutinin protein in parallel with lipids on the cell surface.  This approach may enable evaluating whether distinct proteins are co-localized with specific lipid species in the plasma membrane.