Modulares STED-Mikroskop

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Fluorescence microscopes have been limited for decades by the diffraction barrier, to approximately half of the wavelength of the imaging light (in practice, 200-350 nm). Two types of approaches have been developed to overcome the diffraction barrier. First, the coordinate-targeted approach, which uses a patterned light beam to determine the coordinates from which fluorophores are permitted to emit, used by concepts such as STED. Second, the single-molecule based approach, which is based on the determination of the positions of single fluorophores, which are allowed to emit randomly. This approach is typical of concepts such as PALM, STORM, dSTORM, and GSDIM. STED is faster than PALM/STORM/GSDIM, and is easier to use in live imaging. PALM/STORM/GSDIM are slower, but provide a higher resolution, and are inherently able to count molecules. We propose to combine here the advantages of the two techniques. We propose to purchase the modules that will allow us to build a STED-STORM microscope. This setup combines the advantages of STED, such as high speed, ease of analysis and low propensity to analysis-induced problems, with those of dSTORM or PALM: very high resolution and molecule counting. This will be a unique, never before realized setup, which will open new fields of super-resolution imaging.