Single-Molecule Imaging to Characterize the Transport Mechanism of the Nuclear Pore Complex.
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.
Structural & Molecular Biology, University College London, Gower Street, London, WC1E 6BT, UK.
Grace
G
Jeremy
Department of Biological Sciences, Birkbeck College, University of London, Malet Street, London, WC1E 7HX, UK.
James
J
Stevens
Structural & Molecular Biology, University College London, Gower Street, London, WC1E 6BT, UK. a.lowe@ucl.ac.uk.|Department of Biological Sciences, Birkbeck College, University of London, Malet Street, London, WC1E 7HX, UK. a.lowe@ucl.ac.uk.|London Centre for Nanotechnology, 17-19 Gordon Street, London, WC1H 0AH, UK. a.lowe@ucl.ac.uk.
Alan R
AR
Lowe
D002467:Cell Nucleus / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D028861:Nuclear Pore Complex Proteins / Q000378:metabolism*; D021381:Protein Transport; D000072760:Single Molecule Imaging / Q000379:methods*
D028861:Nuclear Pore Complex Proteins
Nuclear pore complex; Nucleus; Single-molecule tracking; Super-resolution microscopy