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Updated: Apr 3, 2026
11:56
Fluorescence Imaging with One-nanometer Accuracy FIONA
Published on: September 26, 2014
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Real-time fluorescence imaging with 20 nm axial resolution
Daniel R Stabley1, Thomas Oh2, Sanford M Simon2
1Department of Chemistry, Emory University, Atlanta, Georgia 30322, USA.
Nature Communications
|September 23, 2015
Summary
We developed a new imaging method to measure nanoscale protein organization in live cells. This technique allows real-time visualization of dynamic cellular structures with nanometer precision.
Area of Science:
- Cell biology
- Biophysics
- Microscopy
Background:
- Studying nanoscale protein organization in live cells, particularly dynamic structures near the plasma membrane, is technically difficult.
- Existing methods often lack the resolution or real-time capabilities to capture dynamic events.
Purpose of the Study:
- To develop a novel imaging technique for real-time, high-resolution measurement of nanoscale cellular structure height.
- To apply this method to visualize dynamic protein organization in live cells.
Main Methods:
- Development of a two-wavelength total internal reflection fluorescence (TIRF) microscopy method.
- Utilizing proteins tagged with two distinct fluorophores to generate an emission ratio.
- Real-time imaging to capture dynamic changes in cellular structures.
Main Results:
- Demonstrated the capability of nanometer-resolution imaging of cellular structure height in real-time.
- Successfully visualized the nanoscale organization of microtubules.
- Observed the endocytosis process of the epidermal growth factor receptor.
Conclusions:
- The developed two-wavelength TIRF method provides a powerful tool for studying nanoscale organization of dynamic cellular structures.
- This technique offers new possibilities for investigating protein localization and dynamics at the plasma membrane.
- Enables real-time, high-resolution insights into key cellular processes like microtubule dynamics and receptor trafficking.
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