Modulares STED-Mikroskop

几十年来，荧光显微镜一直受到衍射障碍的限制，大约是成像光波长的一半(实际上，200-350 nm)。已经开发了两种类型的方法来克服衍射势垒。首先，STED等概念使用的坐标目标法，它使用图案化的光束来确定允许荧光团发射的坐标。第二，基于单分子的方法，它基于单个荧光团的位置的确定，这些荧光团被允许随机发射。这种方法是Palm、Storm、dSTORM和GSDIM等概念的典型代表。STED比Palm/Storm/GSDIM更快，更易于在实时成像中使用。Palm/Storm/GSDIM速度较慢，但提供更高的分辨率，并且本质上能够计算分子。我们建议在这里结合这两种技术的优点。我们计划购买模块，使我们能够建造STED-STORM显微镜。这种设置结合了STED的优势，如高速、易于分析和低分析引发问题的倾向，以及dSTORM或Palm的优势：非常高的分辨率和分子计数。这将是一种独特的、前所未有的设置，它将开启超分辨率成像的新领域。

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.