Modulares STED-Mikroskop

几十年来，荧光显微镜一直受到衍射障碍的限制，大约只有成像光波长的一半（实际上是200-350纳米）。已经开发了两种克服衍射势垒的方法。首先是坐标定向方法，它使用一束有图案的光束来确定允许荧光团发射的坐标，这种方法被诸如STED之类的概念所使用。第二种是基于单分子的方法，这种方法是基于确定单个荧光团的位置，允许它们随机发射。这种方法是典型的概念，如PALM、STORM、dSTORM和GSDIM。STED比PALM/STORM/GSDIM更快，并且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，并且固有地能够计数分子。我们建议在这里结合这两种技术的优点。我们建议购买模块，使我们能够建立一个STED-STORM显微镜。这种设置结合了STED的优点，例如高速度，易于分析和低倾向于分析引起的问题，以及dSTORM或PALM的优点：非常高的分辨率和分子计数。这将是一个独特的，从未实现过的设置，它将打开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.