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Single-Molecule Imaging of Nuclear Transport
Published on: June 9, 2010
Studying DNA translocation in nanocapillaries using single molecule fluorescence
Vivek V Thacker1, Sandip Ghosal, Silvia Hernández-Ainsa
1Cavendish Laboratory, University of Cambridge, JJ Thompson Avenue, Cambridge CB3 0HE, United Kingdom.
Applied Physics Letters
|January 4, 2013
Summary
We simultaneously measured DNA translocation through nanopores using ionic current and fluorescence. This confirmed DNA passage correlates with ionic current blockages, enabling mobility constant calculation.
Area of Science:
- Nanotechnology
- Biophysics
- Molecular Biology
Background:
- Accurate measurement of DNA translocation is crucial for understanding molecular interactions within nanoscale devices.
- Simultaneous detection methods can improve the reliability and information gained from single-molecule experiments.
Purpose of the Study:
- To demonstrate simultaneous ionic current and fluorescent detection of DNA translocation through glass nanopores.
- To correlate ionic current blockages with DNA molecule passage.
- To determine the effective mobility constant of DNA under high electric fields.
Main Methods:
- Utilizing glass nanopores for DNA translocation.
- Employing ionic current detection to monitor molecule passage.
- Using fluorescent imaging to track individual DNA molecules.
- Applying a model to extract mobility constants from motion tracking.
Main Results:
- Successful simultaneous measurement of DNA translocation events.
- Verified direct correspondence between DNA passage and ionic current blockages.
- Extracted an effective mobility constant for DNA in the specified geometry and high electric fields.
Conclusions:
- Simultaneous ionic current and fluorescence detection provides robust data for DNA translocation studies.
- The method allows for precise characterization of DNA dynamics in nanopores.
- This technique offers a pathway for developing advanced nanoscale DNA analysis tools.
