研究目的
To demonstrate that placing the emitter in front of a metal-dielectric nanocoating that acts as a tuned mirror can provide maximized precision and a high signal-to-noise ratio in single-molecule localization microscopy (SMLM).
研究成果
Metal-dielectric coatings significantly improve the localization precision in SMLM by enhancing the photon yield and reducing background noise. This method is compatible with dual-color imaging and can be easily integrated into existing SMLM setups without the need for complex modifications.
研究不足
The enhancement field of mirror-enhanced SMLM is bound to the surface and currently extends to 160 nm above the substrate interface. This limits its application to samples that are within this range from the surface.
1：Experimental Design and Method Selection:
The study employed finite element method simulations to design metal-dielectric substrates for optimal fluorescence enhancement.
2：Sample Selection and Data Sources:
Nuclear pore complexes (NPCs) and Jurkat T-cells were used as samples to demonstrate the enhancement effect.
3：List of Experimental Equipment and Materials:
Metal-dielectric coatings (2 nm Ge, 50 nm Ag, and 10 nm Si3N4) on glass coverslips, Alexa Fluor 647 (A647) and Alexa Fluor 532 (A532) dyes.
4：Experimental Procedures and Operational Workflow:
dSTORM experiments were performed on coated and uncoated coverslips, with comparisons made between sunny-side-down (SSD) and TIRF configurations.
5：Data Analysis Methods:
Fourier ring correlation (FRC) analysis and radial-aperture-based temporal, intensity photometric estimation (TRABI) were used to analyze the data.
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