ArticleMethods (San Diego, Calif.)2006
Visualizing single molecules interacting with nuclear pore complexes by narrow-field epifluorescence microscopy.
Article in Methods (San Diego, Calif.), 2006. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 20 papers.
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Who cites it
20 citing papers in PubMed.
- Clustering within a single-component biomolecular condensate.bioRxiv : the preprint server for biology · 2025Article
- High-speed super-resolution imaging of rotationally symmetric structures using SPEED microscopy and 2D-to-3D transformation.Nature protocols · 2021Article
- Article
- Time-resolved biophysical approaches to nucleocytoplasmic transport.Computational and structural biotechnology journal · 2017Review
- Deciphering the Structure and Function of Nuclear Pores Using Single-Molecule Fluorescence Approaches.Journal of molecular biology · 2016Review
- Super-resolution 3D tomography of interactions and competition in the nuclear pore complex.Nature structural & molecular biology · 2016Article
- Effect of time discretization of the imaging process on the accuracy of trajectory estimation in fluorescence microscopy.Optics express · 2014Article
- Quantitation of glucocorticoid receptor DNA-binding dynamics by single-molecule microscopy and FRAP.PloS one · 2014Article
- Large cargo transport by nuclear pores: implications for the spatial organization of FG-nucleoporins.The EMBO journal · 2013Article
- Choreography of importin-α/CAS complex assembly and disassembly at nuclear pores.Proceedings of the National Academy of Sciences of the United States of America · 2013Article
- Self-regulated viscous channel in the nuclear pore complex.Proceedings of the National Academy of Sciences of the United States of America · 2012Article
- Single-molecule studies of nucleocytoplasmic transport: from one dimension to three dimensions.Integrative biology : quantitative biosciences from nano to macro · 2012Review
- Single molecule studies of nucleocytoplasmic transport.Biochimica et biophysica acta · 2011Review
- Fluorescence correlation spectroscopy of intact nuclear pore complexes.Biophysical journal · 2011Article
- Calcium regulation of nucleocytoplasmic transport.Protein & cell · 2011Review
- Single-molecule imaging of nuclear transport.Journal of visualized experiments : JoVE · 2010Article
- Three-dimensional distribution of transient interactions in the nuclear pore complex obtained from single-molecule snapshots.Proceedings of the National Academy of Sciences of the United States of America · 2010Article
- Single-molecule measurements of importin alpha/cargo complex dissociation at the nuclear pore.Proceedings of the National Academy of Sciences of the United States of America · 2008Article
- Nuclear import time and transport efficiency depend on importin beta concentration.The Journal of cell biology · 2006Article
- 'Natively unfolded' nucleoporins in nucleocytoplasmic transport: clustered or evenly distributed?Nucleus (Austin, Tex.)Article
Corrections and comments
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2 authors.
Funding
Abstract
The utility of single molecule fluorescence (SMF) for understanding biological reactions has been amply demonstrated by a diverse series of studies over the last decade. In large part, the molecules of interest have been limited to those within a small focal volume or near a surface to achieve the high sensitivity required for detecting the inherently weak signals arising from individual molecules. Consequently, the investigation of molecular behavior with high time and spatial resolution deep within cells using SMF has remained challenging. Recently, we demonstrated that narrow-field epifluorescence microscopy allows visualization of nucleocytoplasmic transport at the single cargo level. We describe here the methodological approach that yields 2 ms and approximately 15 nm resolution for a stationary particle. The spatial resolution for a mobile particle is inherently worse, and depends on how fast the particle is moving. The signal-to-noise ratio is sufficiently high to directly measure the time a single cargo molecule spends interacting with the nuclear pore complex. Particle tracking analysis revealed that cargo molecules randomly diffuse within the nuclear pore complex, exiting as a result of a single rate-limiting step. We expect that narrow-field epifluorescence microscopy will be useful for elucidating other binding and trafficking events within cells.
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