首页> 外国专利> EVALUATION OF SIGNALS OF FLUORESCENCE SCANNING MICROSCOPY USING A CONFOCAL LASER SCANNING MICROSCOPE

EVALUATION OF SIGNALS OF FLUORESCENCE SCANNING MICROSCOPY USING A CONFOCAL LASER SCANNING MICROSCOPE

机译:使用共聚焦激光扫描显微镜评估荧光扫描显微镜的信号

摘要

A method for evaluating signals of fluorescence scanning microscopy with simultaneous excitation and detection of fluorescence in different focal planes of a sample by means of confocal laser scanning microscopy. The invention evaluates signals of fluorescence scanning microscopy without the signal losses usually taking place with a confocal aperture, by coupling an illumination beam into a microscope observation beam path which images a measuring volume on a detector array arranged in the image plane, focusing the illumination beam which passes through a beam-forming phase mask for generating an elongated focus in the measuring volume, collecting and collimating fluorescent light generated in the measuring volume and routing it to diffractive optics which split the light beams into different diffraction orders and impress a different spherical phase on the light beams, imaging the different diffraction orders on detector regions of the detector array so that fluorescent light from focal planes at different depths of the measuring volume are associated with different diffraction orders, and associating the fluorescence signals on which crosstalk is superposed from different focal planes of the measuring volume with defined focal planes by means of correlation-based association based on distinguishable blinking behavior of fluorescing dyes.
机译:一种通过共聚焦激光扫描显微镜同时激发和检测样品不同焦平面中的荧光来评估荧光扫描显微镜信号的方法。本发明通过将照明束耦合到显微镜观察束路径中来评估荧光扫描显微镜的信号,而该信号通常不会在共焦孔径下发生信号损失,该显微镜观察束路径将测量体积成像在布置在图像平面中的检测器阵列上,聚焦照明束它穿过光束形成相位掩模,以在测量空间中产生细长的焦点,收集并准直在测量空间中产生的荧光,并将其路由到衍射光学器件,该光学器件将光束分成不同的衍射级并施加不同的球形相位在光束上,对检测器阵列的检测器区域上不同的衍射级进行成像,以使来自测量体积不同深度的焦平面的荧光与不同的衍射级相关联,并将其上叠加有不同串扰的荧光信号相关联的焦平面通过基于相关性的关联基于荧光染料的明显闪烁行为测量具有定义的焦平面的体积。

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