Related Experiment Video
Updated: Jun 12, 2026
12:13
Single-Molecule Imaging of Nuclear Transport
Published on: June 9, 2010
Single-molecule imaging of nuclear transport
Alexander Goryaynov1, Ashapurna Sarma, Jiong Ma
1Department of Biology, Bowling Green State University, USA.
Journal of Visualized Experiments : Jove
|June 16, 2010
Summary
This study enhances single molecule fluorescence microscopy for visualizing molecular transport within cells. The improved method captures rapid transport events through the nuclear pore complex (NPC) with unprecedented resolution.
Area of Science:
- Cell Biology
- Biophysics
- Microscopy
Background:
- Single molecule fluorescence microscopy (SMFM) is crucial for studying biological reactions.
- Investigating molecular interactions deep within cells is challenging due to weak signals.
- Previous methods visualized nucleocytoplasmic transport but lacked high spatiotemporal resolution.
Purpose of the Study:
- To describe a protocol for an improved SMFM method.
- To achieve enhanced spatiotemporal resolution for capturing cellular transport events.
- To enable detailed studies of molecular trafficking within cells.
Main Methods:
- Utilized narrow-field epifluorescence microscopy.
- Developed a protocol achieving 0.4 ms temporal and 12 nm spatial resolution.
- Applied the method to semi-intact cells to study nuclear transport.
Main Results:
- Achieved unprecedented spatiotemporal resolution in SMFM.
- Successfully visualized nucleocytoplasmic transport of cargo molecules.
- Captured transient active transport and passive diffusion events through nuclear pore complexes (NPCs).
Conclusions:
- The developed SMFM protocol offers significant improvements in resolution.
- This method facilitates the study of rapid molecular transport and diffusion.
- The technique is expected to advance research on intracellular binding and trafficking events.
Related Concept Videos
01:42Directionality of Nuclear Transport
Ras-related nuclear protein or Ran is a small G protein that cycles between its GTP and GDP bound states. Ran specific regulators, a Ran GTPase Activating Protein or RanGAP present in the cytosol and a Ran guanine nucleotide exchange factor or RanGEF present inside the nucleus regulate GTP/GDP exchange. A high concentration of GTP inside the cells, in addition to this asymmetric distribution of Ran-specific regulators, leads to a higher RanGTP concentration inside the nucleus. This...
01:34Nuclear Protein Sorting
Nuclear protein sorting is the selective trafficking of histones, polymerases, gene regulatory proteins into the nucleus and exporting RNAs and ribosomes to the cytosol. It is a tightly controlled process that regulates gene expression within a cell.
Proteins targeted to the nucleus carry nuclear localization signals or NLS recognized by import receptors in the cytosol. Similarly, proteins with nuclear export signals are recognized by export receptors. Import and export receptors are...
Proteins targeted to the nucleus carry nuclear localization signals or NLS recognized by import receptors in the cytosol. Similarly, proteins with nuclear export signals are recognized by export receptors. Import and export receptors are...
01:42Nuclear Export
The nucleus restricts several proteins within and allows others to pass. The restricted proteins possess a nuclear retention sequence or NRS, anchoring them to the nuclear lamins and preventing their transport to the cytosol. The non-restricted proteins, after their synthesis, are transported to their site of action, such as the cytosol or other organelles, with the help of nuclear export signals or NES.
NES are of three types- the canonical 10-residue long leucine-rich signal and other...
NES are of three types- the canonical 10-residue long leucine-rich signal and other...