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Monitoring Conformational Dynamics of Single Unmodified Proteins using Plasmonic Nanotweezers
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Fluorescence axial nanotomography with plasmonics
Nicholas I Cade1, Gilbert O Fruhwirth, Alexey V Krasavin
1Department of Physics, King's College London, Strand, London WC2R 2LS, UK. david.r.richards@kcl.ac.uk.
Faraday Discussions
|February 26, 2015
Summary
We developed a new super-resolution imaging method using fluorescence lifetime changes near plasmonic substrates. This technique achieves 6 nm axial sensitivity for imaging cellular structures and processes like endocytosis.
Area of Science:
- Biophysics
- Optical Imaging
- Cell Biology
Background:
- Conventional imaging techniques lack sufficient axial resolution for detailed cellular studies.
- Understanding cellular membrane topography is crucial for studying biological processes.
Purpose of the Study:
- To introduce a novel super-resolution imaging technique with enhanced axial sensitivity.
- To demonstrate the application of this technique in mapping cellular membrane topography and studying endocytosis.
Main Methods:
- Utilizing changes in fluorescence lifetime above a plasmonic substrate.
- Employing conventional confocal fluorescence lifetime imaging.
- Achieving down to 6 nm axial position sensitivity of fluorophores.
Main Results:
- Demonstrated 6 nm axial position sensitivity in whole biological cell imaging.
- Successfully mapped the topography of the cellular membrane.
- Applied the technique to investigate receptor-mediated endocytosis in carcinoma cells.
Conclusions:
- The novel technique offers unprecedented axial resolution for cellular imaging.
- This method provides a powerful tool for studying membrane dynamics and molecular interactions.
- It has significant potential for advancing research in cell biology and disease mechanisms.
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