研究目的
Investigating the therapeutic effects of a specific herbal medicine on a particular disease.
研究成果
The study demonstrated that graphene oxide films combined with STORM imaging provide a convenient method for imaging samples with large backgrounds due to non-specifically bound fluorophores, offering significant improvements in contrast and resolution. The technique's applicability to a variety of samples and its potential for extension to other fluorescence microscopy methods were highlighted.
研究不足
The study's limitations include the potential for variation in quenching efficiency due to the non-flat nature of graphene oxide sheets and thickness variations in spin-coated polymer layers. Additionally, the resolution improvements, while significant, were sample and metric dependent.
1:Experimental Design and Method Selection:
The study employed the dSTORM variant of STORM microscopy, utilizing photoswitchable fluorophores to separate individual fluorescence signals by stochastic blinking. The position of each localization was determined by fitting a two-dimensional Gaussian to the point spread function (PSF) of each fluorophore’s signal.
2:Sample Selection and Data Sources:
Samples included individual sheets of graphene oxide, self-assembled peptide aggregates, and live bacterial cells. Graphene oxide layers were spin-coated on substrates from aqueous solutions.
3:List of Experimental Equipment and Materials:
A Digital sCMOS camera (Hamamatsu C11440-22CU), a 100x objective lens (UAPON 100XOTIRF, Olympus), and a MadCity Labs C-focus system for sample drift elimination were used. Fluorophores included Alexa Fluor 647 and Cy3B NHS ester.
4:Experimental Procedures and Operational Workflow:
The method involved coating substrates with graphene oxide, applying polymer spacer layers, and then fluorophores. Imaging was performed with STORM, and data was analyzed with ThunderSTORM.
5:Data Analysis Methods:
The Fourier Ring Correlation method was used to calculate image resolutions, and the energy transfer efficiency was evaluated using photon densities from quenched and non-quenched areas.
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