Endocytic pathway revisited by single molecule tracking

FA Meunier and M Joensuu

Queensland Brain Institute. The University of Queensland

The diffraction limit of light microscopy severely hampers our understanding of the dynamics of the endocytic pathway. In this presentation we introduce a novel technique based on the sub-diffractional tracking of internalized molecules (sdTIM) to image anti-GFP Atto647N-tagged nanobodies trapped in synaptic vesicles (SVs) with 50 nm localization precision. Using Bayesian model selection applied to hidden Markov modeling we found that SVs constantly oscillated between one immobile and two transport states of opposite direction. These results highlight the potential of the sdTIM technique to provide new dynamic insights into endocytic pathways in various cellular settings.