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Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells
Published on: June 30, 2018
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Electrochemically modulated single-molecule localization microscopy for in vitro imaging cytoskeletal protein
1College of Chemistry and Bioengineering, Guilin University of Technology, Guilin, Guangxi 541006, China.
Nanophotonics (Berlin, Germany)
|February 20, 2025
Summary
A novel electrochemical method enhances super-resolution microscopy by controlling fluorophore blinking. This technique allows imaging with more molecules, even those with poor photophysical properties, expanding single-molecule localization microscopy applications.
Area of Science:
- Biophysics
- Chemical Imaging
- Super-resolution Microscopy
Background:
- Single-molecule localization microscopy (SMLM) is limited by fluorophore duty cycles.
- Unfavorable photophysical properties restrict the use of many potential fluorophores in SMLM.
Purpose of the Study:
- To develop a new concept for electrochemically modulated SMLM.
- To overcome limitations imposed by fluorophore duty cycles in SMLM.
Main Methods:
- Utilized redox-active cresyl violet as a model fluorophore with an unfavorable high duty cycle.
- Synchronized electrochemical potential scanning with SMLM to control fluorophore On/Off states.
- Applied the technique to image cytoskeletal proteins (microtubules and actins) in vitro.
Main Results:
- Successfully modulated the On/Off states of single redox-active fluorophores using electrochemical potential.
- Obtained super-resolution images of microtubules and actins via precise single-molecule localizations.
- Demonstrated the feasibility of using fluorophores with unfavorable photophysical properties.
Conclusions:
- Electrochemical modulation offers a new strategy to improve SMLM.
- This method expands the range of usable fluorophores, enhancing SMLM's applicability.
- Enables wider and more extensive applications of single-molecule localization microscopy.
