Abstract
Single molecule tracking in live cells is the ultimate tool to study subcellular protein dynamics, but it is often limited by the probe size and photostability. Due to these issues, long-term tracking of proteins in confined and crowded environments, such as adhesion sites, synaptic clefts or intracellular spaces, remains challenging. We present a novel optical probe consisting of 5-nm gold nanoparticles functionalized with a small fragment of camelid antibodies that recognize widely used GFPs with a very high affinity (1). These small gold nanoparticles can be detected and tracked using photothermal imaging for arbitrarily long periods of time (1-2). Surface and intracellular GFP-proteins can effectively be labeled even in very crowded environments such as adhesion sites and cytoskeletal structures both in vitro and in live cell cultures. Comparison with performances obtained by superresolution methods such as PALM and STED are presented for single integrin tracking in and out adhesion sites (3). These nanobody-coated gold nanoparticles are single molecule probes with unparalleled capabilities; small size, perfect photostability, high specificity, and versatility afforded by combination with the vast existing library of GFP-tagged proteins.
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