Single-Molecule Imaging to Characterize the Transport Mechanism of the Nuclear Pore Complex

  • Grace Jeremy
  • James Stevens
  • Alan R. LoweEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1431)


In the eukaryotic cell, a large macromolecular channel, known as the Nuclear Pore Complex (NPC), mediates all molecular transport between the nucleus and cytoplasm. In recent years, single-molecule fluorescence (SMF) imaging has emerged as a powerful tool to study the molecular mechanism of transport through the NPC. More recently, techniques such as single-molecule localization microscopy (SMLM) have enabled the spatial and temporal distribution of cargos, transport receptors and even structural components of the NPC to be determined with nanometre accuracy. In this protocol, we describe a method to study the position and/or motion of individual molecules transiting through the NPC with high spatial and temporal precision.

Key words

Nucleus Nuclear pore complex Single-molecule tracking Super-resolution microscopy 



Grace Jeremy is supported by a Wellcome Trust studentship. We thank Anthony Roberts for criticial reading of the manuscript. We also thank the Hayward, Waksman and Fassati labs for contributions of reagents, equipment, and expertise. The Lowe lab acknowledges support from the Medical Research Council award MR/K015826/1 Super Resolution Imaging for Cell Biology and Neuroscience at UCL.


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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Structural & Molecular BiologyUniversity College LondonLondonUK
  2. 2.Department of Biological Sciences, Birkbeck CollegeUniversity of LondonLondonUK
  3. 3.London Centre for NanotechnologyLondonUK

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