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1. 发明申请

WO2014180566A1 METHOD AND APPARATUS FOR COMBINATION OF LOCALIZATION MICROSCOPY AND STRUCTURED ILLUMINATION MICROSCOPY 审中-公开
标题翻译：本地化显微镜和结构化照明显微镜的组合方法与装置
公开(公告)号：WO2014180566A1
公开(公告)日：2014-11-13
申请号：PCT/EP2014/001228
申请日：2014-05-07
申请人： RUPRECHT-KARLS-UNIVERSITÄT HEIDELBERG
发明人： BEST, Gerrit , CREMER, Christoph , ROSSBERGER, Sabrina , DITHMAR, Stefan
IPC分类号： G02B21/00 , G01N21/64 , G02B21/16 , G02B21/36 , G02B27/58
CPC分类号： G02B21/367 , G01N21/6445 , G01N21/6458 , G01N2201/0675 , G02B21/0076 , G02B21/16 , G02B21/361 , G02B27/58
摘要： The invention relates to a fluorescent microscope and a respective method for obtaining super-resolution images of a sample labelled with at least one type fluorescent label by combining localization microscopy and structured illumination microscopy. In an aspect, the fluorescent microscope comprises one or more light sources and an illumination system having a structured illumination path, in which a pattern generation system is positioned, for illuminating the sample with structured illumination light and a localization illumination path for illuminating the sample with localization illumination light. A switching mechanism is configured to switch between a first, a second and/or a third mode, wherein in the first mode at least a portion of the light emitted from the one or more light sources propagates through one of the illumination paths; in the second mode at least a portion of the light emitted from the one or more light sources propagates through the other one of the illumination paths; and in the third mode at least a portion of the light emitted from one or more of the light sources propagates through one illumination path while simultaneously at least another portion of the light emitted from one or more of the light sources propagates through the other illumination path. At least one image detector positioned in an optical detection path, configured to detect at least a portion of fluorescent light emitted from fluorescent molecules of the illuminated sample. Another aspect concerns a method for obtaining super-resolution image data of a sample labeled with at least one type of fluorescent label comprising illuminating the sample with localization illumination light and with structured illumination light; detecting at least a portion of the fluorescent light emitted from the excited fluorescent molecules of the at least one fluorescent label, thereby obtaining at least one image of the illuminated sample; and processing the obtained at least one image of the sample image to obtain super-resolution image data.
摘要翻译：本发明涉及通过组合定位显微镜和结构化照明显微镜获得用至少一种类型荧光标记物标记的样品的超分辨率图像的荧光显微镜和相应方法。在一方面，荧光显微镜包括一个或多个光源和具有结构化照明路径的照明系统，其中定位图案生成系统，用于以结构化的照明光照射样本，并且具有用于照亮样品的定位照明路径，定位照明灯。切换机构被配置为在第一模式，第二模式和/或第三模式之间切换，其中在第一模式中，从一个或多个光源发射的光的至少一部分传播通过照明路径之一; 在第二模式中，从一个或多个光源发射的光的至少一部分传播通过另一个照明路径; 并且在第三模式中，从一个或多个光源发射的光的至少一部分传播通过一个照明路径，同时，从一个或多个光源发射的光的至少另一部分传播通过另一个照明路径。位于光学检测路径中的至少一个图像检测器，被配置为检测从被照射样品的荧光分子发射的荧光的至少一部分。另一方面涉及一种用于获得用至少一种类型的荧光标签标记的样品的超分辨率图像数据的方法，包括用定位照明光和结构化照明光照射样品; 检测从所述至少一个荧光标记的激发的荧光分子发射的荧光的至少一部分，从而获得所述被照射样品的至少一个图像; 以及处理获得的样本图像的至少一个图像以获得超分辨率图像数据。

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