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Biophys J
2005 May 01;885:3681-8. doi: 10.1529/biophysj.104.051458.
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High-resolution near-field optical imaging of single nuclear pore complexes under physiological conditions.
Höppener C
,
Siebrasse JP
,
Peters R
,
Kubitscheck U
,
Naber A
.
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Scanning near-field optical microscopy (SNOM) circumvents the diffraction limit of conventional light microscopy and is able to achieve optical resolutions substantially below 100 nm. However, in the field of cell biology SNOM has been rarely applied, probably because previous techniques for sample-distance control are less sensitive in liquid than in air. Recently we developed a distance control based on a tuning fork in tapping mode, which is also well-suited for imaging in solution. Here we show that this approach can be used to visualize single membrane protein complexes kept in physiological media throughout. Nuclear envelopes were isolated from Xenopus laevis oocytes at conditions shown recently to conserve the transport functions of the nuclear pore complex (NPC). Isolated nuclear envelopes were fluorescently labeled by antibodies against specific proteins of the NPC (NUP153 and p62) and imaged at a resolution of approximately 60 nm. The lateral distribution of epitopes within the supramolecular NPC could be inferred from an analysis of the intensity distribution of the fluorescence spots. The different number densities of p62- and NUP153-labeled NPCs are determined and discussed. Thus we show that SNOM opens up new possibilities for directly visualizing the transport of single particles through single NPCs and other transporters.
Adam,
Nuclear protein import in permeabilized mammalian cells requires soluble cytoplasmic factors.
1990, Pubmed,
Xenbase
Adam,
Nuclear protein import in permeabilized mammalian cells requires soluble cytoplasmic factors.
1990,
Pubmed
,
Xenbase
Adam,
The nuclear pore complex.
2001,
Pubmed
Allen,
The nuclear pore complex: mediator of translocation between nucleus and cytoplasm.
2000,
Pubmed
Ben-Efraim,
Gradient of increasing affinity of importin beta for nucleoporins along the pathway of nuclear import.
2001,
Pubmed
Betzig,
Breaking the diffraction barrier: optical microscopy on a nanometric scale.
1991,
Pubmed
Betzig,
Single molecules observed by near-field scanning optical microscopy.
1993,
Pubmed
Chook,
Karyopherins and nuclear import.
2001,
Pubmed
Conti,
Nucleocytoplasmic transport enters the atomic age.
2001,
Pubmed
Dunn,
Near-field scanning optical microscopy.
1999,
Pubmed
Evans,
Biochemical fractionation of oocytes.
1991,
Pubmed
,
Xenbase
Fahrenkrog,
The nuclear pore complex: nucleocytoplasmic transport and beyond.
2003,
Pubmed
Fahrenkrog,
Nuclear pore complex architecture and functional dynamics.
2001,
Pubmed
Fahrenkrog,
Domain-specific antibodies reveal multiple-site topology of Nup153 within the nuclear pore complex.
2002,
Pubmed
,
Xenbase
Fahrenkrog,
Molecular architecture of the yeast nuclear pore complex: localization of Nsp1p subcomplexes.
1998,
Pubmed
,
Xenbase
Gheber,
Design and optimization of a near-field scanning optical microscope for imaging biological samples in liquid.
1998,
Pubmed
Goulian,
Tracking single proteins within cells.
2000,
Pubmed
Höppener,
Scanning near-field optical microscopy of a cell membrane in liquid.
2003,
Pubmed
,
Xenbase
Hu,
Molecular and functional characterization of the p62 complex, an assembly of nuclear pore complex glycoproteins.
1996,
Pubmed
Hwang,
Domains in cell plasma membranes investigated by near-field scanning optical microscopy.
1998,
Pubmed
Keller,
Optical chemical imaging of tobacco mosaic virus in solution at 60-nm resolution.
1998,
Pubmed
Keminer,
Permeability of single nuclear pores.
1999,
Pubmed
,
Xenbase
Kubitscheck,
Single nuclear pores visualized by confocal microscopy and image processing.
1996,
Pubmed
Kues,
High intranuclear mobility and dynamic clustering of the splicing factor U1 snRNP observed by single particle tracking.
2001,
Pubmed
,
Xenbase
Macara,
Transport into and out of the nucleus.
2001,
Pubmed
Naber,
The tuning fork as sensor for dynamic force distance control in scanning near-field optical microscopy.
1999,
Pubmed
Peters,
Optical single transporter recording: transport kinetics in microarrays of membrane patches.
2003,
Pubmed
Rakowska,
ATP-Induced shape change of nuclear pores visualized with the atomic force microscope.
1998,
Pubmed
,
Xenbase
Ribbeck,
Kinetic analysis of translocation through nuclear pore complexes.
2001,
Pubmed
Rout,
The nuclear pore complex as a transport machine.
2001,
Pubmed
Rout,
The yeast nuclear pore complex: composition, architecture, and transport mechanism.
2000,
Pubmed
Ryan,
The nuclear pore complex: a protein machine bridging the nucleus and cytoplasm.
2000,
Pubmed
Seisenberger,
Real-time single-molecule imaging of the infection pathway of an adeno-associated virus.
2001,
Pubmed
Siebrasse,
Reconstitution of nuclear protein export in isolated nuclear envelopes.
2002,
Pubmed
,
Xenbase
Smith,
Oogenesis and oocyte isolation.
1991,
Pubmed
,
Xenbase
Stoffler,
Calcium-mediated structural changes of native nuclear pore complexes monitored by time-lapse atomic force microscopy.
1999,
Pubmed
,
Xenbase
Stoffler,
The nuclear pore complex: from molecular architecture to functional dynamics.
1999,
Pubmed
Vasu,
Nuclear pores and nuclear assembly.
2001,
Pubmed
Walther,
The nucleoporin Nup153 is required for nuclear pore basket formation, nuclear pore complex anchoring and import of a subset of nuclear proteins.
2001,
Pubmed
,
Xenbase
