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Choosing dyes for cw-STED nanoscopy using self-assembled nanorulers
Susanne Beater1, Phil Holzmeister, Enrico Pibiri
1NanoBioSciences Group, Institute for Physical and Theoretical Chemistry, TU Braunschweig, Hans-Sommer-Str. 10, 38106 Braunschweig, Germany. b.lalkens@tu-bs.de p.tinnefeld@tu-bs.de.
Physical Chemistry Chemical Physics : PCCP
|March 7, 2014
Summary
Choosing the right fluorescent dye is crucial for high-quality superresolution microscopy. This study found that optimal dye performance in continuous wave STED microscopy depends on the sample
Area of Science:
- Optical Imaging
- Biophysics
- Materials Science
Background:
- Superresolution microscopy significantly enhances optical imaging quality.
- Sample labeling is critical for achieving high-resolution images.
- Fluorescent dye selection impacts imaging performance.
Purpose of the Study:
- Compare four fluorescent dyes (Abberior Star 488, Alexa 488, Chromeo 488, Oregon Green 488) for continuous wave STED microscopy.
- Evaluate dye performance in aqueous buffer and polymer matrices.
- Assess the utility of DNA origami nanorulers for precise microscopy calibration.
Main Methods:
- Utilized continuous wave STED microscopy.
- Employed DNA origami nanorulers with precisely positioned fluorescent dyes.
- Tested dyes in both aqueous buffer and a polymer embedding medium.
- Analyzed image quality and resolution based on dye and medium.
Main Results:
- All tested dyes (Abberior Star 488, Alexa 488, Chromeo 488, Oregon Green 488) are suitable for cw-STED microscopy.
- Optimal fluorescent dye performance is dependent on the embedding medium.
- DNA origami nanorulers provide accurate distance measurements (∼100 nm) for calibration.
Conclusions:
- Fluorescent dye choice is critical and medium-dependent for optimal STED microscopy.
- DNA origami nanorulers are mature, reliable tools for fluorescence microscopy calibration.
- Advancements in sample labeling and calibration tools drive superresolution imaging progress.
