Protocol

Chromosome Architecture

Volume 1431 of the series Methods in Molecular Biology pp 17-35

Date:

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

  • Grace JeremyAffiliated withStructural & Molecular Biology, University College London
  • , James StevensAffiliated withDepartment of Biological Sciences, Birkbeck College, University of London
  • , Alan R. LoweAffiliated withStructural & Molecular Biology, University College LondonDepartment of Biological Sciences, Birkbeck College, University of LondonLondon Centre for Nanotechnology Email author 

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Abstract

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