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1. 发明申请

US20090250632A1 Method and Arrangement for Collimated Microscopic Imaging 审中-公开
标题翻译：准直显微镜成像的方法和布置
公开(公告)号：US20090250632A1
公开(公告)日：2009-10-08
申请号：US12444290
申请日：2007-10-02
申请人： Michael Kempe , Gerhard Krampert , Matthias Wald , Ralf Wolleschensky
发明人： Michael Kempe , Gerhard Krampert , Matthias Wald , Ralf Wolleschensky
IPC分类号： G01N21/64
CPC分类号： G02B21/16 , G01N21/6458 , G02B21/0032 , G02B21/0072 , G02B21/0076
摘要： A method and arrangement for collimated microscopic imaging, including a first illumination of a sample in at least one region for exciting fluorescence, and a spatially resolving detection of the sample light by detector elements, the detection being associated with the region, wherein by means of a second illumination a sub-division of the region into separate fluorescent partial regions occurs, which are associated with the detector elements. The separation of the partial regions is carried out by the spatial separation of the fluorescent regions by means of intermediate regions having reduced fluorescence or no fluorescence, and/or by means of different spectral properties of the fluorescence from the partial regions.
摘要翻译：一种用于准直显微镜成像的方法和装置，包括用于激发荧光的至少一个区域中的样品的第一照明，以及由检测器元件对样品光进行空间分辨检测，所述检测与所述区域相关联，其中通过发生与检测器元件相关联的区域的分割成分离的荧光部分区域的第二照明。部分区域的分离是通过荧光区域通过具有减少的荧光或无荧光的中间区域和/或通过来自部分区域的荧光的不同光谱性质的空间分离来进行的。

2. 发明授权

US08207510B2 Method and arrangement for collimated microscopic imaging 有权
标题翻译：准直显微镜成像的方法和布置
公开(公告)号：US08207510B2
公开(公告)日：2012-06-26
申请号：US13042137
申请日：2011-03-07
申请人： Michael Kempe , Gerhard Krampert , Matthias Wald , Ralf Wolleschensky
发明人： Michael Kempe , Gerhard Krampert , Matthias Wald , Ralf Wolleschensky
IPC分类号： G01J1/58
CPC分类号： G02B21/16 , G01N21/6458 , G02B21/0032 , G02B21/0072 , G02B21/0076
摘要： A method and arrangement for collimated microscopic imaging, including a first illumination of a sample in at least one region for exciting fluorescence, and a spatially resolving detection of the sample light by detector elements, the detection being associated with the region, wherein by means of a second illumination a sub-division of the region into separate fluorescent partial regions occurs, which are associated with the detector elements. The separation of the partial regions is carried out by the spatial separation of the fluorescent regions by means of intermediate regions having reduced fluorescence or no fluorescence, and/or by means of different spectral properties of the fluorescence from the partial regions.
摘要翻译：一种用于准直显微镜成像的方法和装置，包括用于激发荧光的至少一个区域中的样品的第一照明，以及由检测器元件对样品光进行空间分辨检测，所述检测与所述区域相关联，其中通过发生与检测器元件相关联的区域的分割成分离的荧光部分区域的第二照明。部分区域的分离是通过荧光区域通过具有减少的荧光或无荧光的中间区域和/或通过来自部分区域的荧光的不同光谱性质的空间分离来进行的。

3. 发明授权

US08908174B2 Apparatus, especially microscope, for the analysis of samples 有权
标题翻译：仪器，特别是显微镜，用于分析样品
公开(公告)号：US08908174B2
公开(公告)日：2014-12-09
申请号：US13121919
申请日：2009-09-22
申请人： Michael Kempe , Gerhard Krampert , Ingo Kleppe , Ralf Wolleschensky
发明人： Michael Kempe , Gerhard Krampert , Ingo Kleppe , Ralf Wolleschensky
IPC分类号： G01J3/28 , G02B21/16 , G02B21/00 , G01N21/64
CPC分类号： G01N21/6428 , G01N21/6458 , G01N2021/6417 , G02B21/0076 , G02B21/16
摘要： A microscope device which has a diffraction-limited resolution volume, with multiple dye molecules that can be switched between different states, at least one of which is fluorescent. The fluorescence is focused using an objective lens and is imaged onto a spatially resolving detector. In at least one portion of the sample, the dye molecules have a distribution density that is greater than the inverse of the diffraction-limited resolution volume. One or more light sources are provided for emitting a switching radiation in order to switch a first subset of the dye molecules in the sample, and for emitting an excitation radiation in order to excite the first subset of dye molecules. A phase mask which generates a light distribution having an at least partially limited local minimum radiation on the detector plane is provided in the beam path, preferably in the detection beam path.
摘要翻译：具有衍射限制分辨率体积的显微镜装置，具有可以在不同状态之间切换的多个染料分子，其中至少一个是荧光的。使用物镜聚焦荧光，并将其成像到空间分辨检测器上。在样品的至少一部分中，染料分子的分布密度大于衍射极限分辨率体积的倒数。提供一个或多个光源用于发射切换辐射，以便切换样品中的染料分子的第一子集，并且用于发射激发辐射以激发染料分子的第一子集。优选在检测光束路径中，在光束路径中设置产生具有至少部分受限局部最小辐射的光分布的相位掩模。

4. 发明授权

US08704196B2 Combination microscopy 有权
标题翻译：组合显微镜
公开(公告)号：US08704196B2
公开(公告)日：2014-04-22
申请号：US13127427
申请日：2009-10-28
申请人： Ralf Wolleschensky , Ingo Kleppe , Gerhard Krampert , Michael Kempe
发明人： Ralf Wolleschensky , Ingo Kleppe , Gerhard Krampert , Michael Kempe
IPC分类号： G01J1/58
CPC分类号： G02B27/642 , G01N21/6458 , G02B21/0076 , G02B21/367 , G02B27/58
摘要： A method for generating an image of a sample by a microscopy method including varying local resolution, wherein at least two of the following microscopy methods are combined: laser scanning microscopy, a microscopy method wherein the sample is excited to luminescence by structured line or wide area illumination, and a first microscopy image is generated from the images thus obtained, having increased local resolution greater than the optical resolution of the image, a further microscopy method according to the PAL principle, by which a second microscopy image is generated, indicating geometric locations of marker molecules emitting luminescent radiation at an increased local resolution relative to the optical resolution, and a further microscopy method, wherein the sample is marked using marking molecules suitable for the STED, ESA, or RESOLFT technique, and a third microscopy image is generated of STED, ESA, or RESOLFT, wherein the obtained images are superimposed.
摘要翻译：一种用于通过包括变化的局部分辨率的显微镜方法产生样品的图像的方法，其中组合以下显微镜方法中的至少两种：激光扫描显微镜，其中通过结构化线或广泛区域将样品激发至发光的显微镜方法照明，并且从由此获得的图像产生第一显微镜图像，其具有大于图像的光学分辨率的增加的局部分辨率，根据PAL原理的另一显微镜方法，通过该方法生成第二显微镜图像，指示几何位置相对于光学分辨率以增加的局部分辨率发射发光辐射的标记分子，以及另外的显微镜方法，其中使用适合于STED，ESA或RESOLFT技术的标记分子标记样品，并且产生第三显微镜图像 STED，ESA或RESOLFT，其中所获得的图像被叠加。

5. 发明授权

US08610086B2 Increased resolution microscopy 有权
标题翻译：增加分辨率显微镜
公开(公告)号：US08610086B2
公开(公告)日：2013-12-17
申请号：US13131801
申请日：2009-11-14
申请人： Ralf Wolleschensky , Ingo Kleppe , Gerhard Krampert , Michael Kempe
发明人： Ralf Wolleschensky , Ingo Kleppe , Gerhard Krampert , Michael Kempe
IPC分类号： G01N21/64
CPC分类号： G02B21/16 , G01N21/6458 , G02B21/361 , G02B21/367 , G02B27/58
摘要： Method for spatially high-resolution luminescence microscopy in which label molecules in a sample are activated to emit luminescence radiation comprising activating only a subset of the label molecules in the sample, wherein activated label molecules have a distance to the closest activated molecules that is greater or equal to a length which results from a predetermined optical resolution, detecting the luminescence radiation, generating a frame from the luminescence radiation, identifying the geometric locations of the label molecules with a spatial resolution increased above the predetermined optical resolution, repeating the steps and forming a combined image, and controlling the acquisition of the several frames by evaluating at least one of the frames or a group of the frames and modifying at least one variable for subsequent repetitions of the steps of generating frames for combining into an image.
摘要翻译：用于空间高分辨率发光显微镜的方法，其中样品中的标记分子被激活以发射发光，包括仅激活样品中标记分子的一个子集，其中激活的标记分子与最接近的激活分子的距离更大或等于由预定的光学分辨率产生的长度，检测发光辐射，从发光辐射产生帧，以高于预定的光学分辨率的空间分辨率识别标签分子的几何位置，重复步骤并形成并且通过评估所述帧或一组帧中的至少一个并且修改至少一个变量来控制所述几帧的获取，以便后续重复生成用于组合成图像的帧的步骤。

6. 发明申请

US20110182529A1 METHODS AND APPARATUSES FOR STRUCTURED ILLUMINATION MICROSCOPY 有权
标题翻译：用于结构化照明显微镜的方法和装置
公开(公告)号：US20110182529A1
公开(公告)日：2011-07-28
申请号：US13121466
申请日：2009-09-22
申请人： Michael Kempe , Gerhard Krampert , Ingo Kleppe , Ralf Wolleschensky
发明人： Michael Kempe , Gerhard Krampert , Ingo Kleppe , Ralf Wolleschensky
IPC分类号： G06K9/40 , G01J1/58
CPC分类号： G02B21/0032 , G01N21/6458 , G02B21/0072 , G02B21/0076 , G02B21/008 , G02B21/367 , G02B27/58
摘要： In structured illumination microscopy, the multiple recording of images with different phase positions of the structuring requires a high stability in the optical arrangement and sample throughout the entire measuring process. Also, the structuring must be projected into the sample in a highly homogeneous manner. The current invention optimizes recording of individual images in order to achieve the best possible resolution in the result image even in problematic samples. An optimization of this kind can be carried out in different ways, for example, by determining an optimal adjustment for at least one illumination parameter or recording parameter or by pulsed illumination such that an excitation from a triplet state of the fluorescent dye to a higher triplet state is reduced, or by illuminating the sample with depletion light for depopulating a triplet state of the fluorescent dye, which reduces bleaching.
摘要翻译：在结构化照明显微镜中，具有结构化不同相位位置的图像的多次记录在整个测量过程中需要光学布置和样品中的高稳定性。此外，结构化必须以高度均匀的方式投射到样品中。本发明优化各个图像的记录，以便即使在有问题的样本中也能在结果图像中实现最佳分辨率。可以以不同的方式进行这种优化，例如通过确定至少一个照明参数或记录参数的最佳调整或通过脉冲照明使得从荧光染料的三重态到更高三重态的激发或通过用耗尽光照射样品来减少荧光染料的三线态，从而减少漂白。

7. 发明授权

US09348127B2 Methods and apparatuses for structured illumination microscopy 有权
标题翻译：结构照明显微镜的方法和装置
公开(公告)号：US09348127B2
公开(公告)日：2016-05-24
申请号：US13121466
申请日：2009-09-22
申请人： Michael Kempe , Gerhard Krampert , Ingo Kleppe , Ralf Wolleschensky
发明人： Michael Kempe , Gerhard Krampert , Ingo Kleppe , Ralf Wolleschensky
IPC分类号： G06K9/40 , G02B21/00 , G01N21/64 , G02B21/36 , G02B27/58
CPC分类号： G02B21/0032 , G01N21/6458 , G02B21/0072 , G02B21/0076 , G02B21/008 , G02B21/367 , G02B27/58
摘要： In structured illumination microscopy, the multiple recording of images with different phase positions of the structuring requires a high stability in the optical arrangement and sample throughout the entire measuring process. Also, the structuring must be projected into the sample in a highly homogeneous manner. The current invention optimizes recording of individual images in order to achieve the best possible resolution in the result image even in problematic samples. An optimization of this kind can be carried out in different ways, for example, by determining an optimal adjustment for at least one illumination parameter or recording parameter or by pulsed illumination such that an excitation from a triplet state of the fluorescent dye to a higher triplet state is reduced, or by illuminating the sample with depletion light for depopulating a triplet state of the fluorescent dye, which reduces bleaching.
摘要翻译：在结构化照明显微镜中，具有结构化不同相位位置的图像的多次记录在整个测量过程中需要光学布置和样品中的高稳定性。此外，结构化必须以高度均匀的方式投射到样品中。本发明优化各个图像的记录，以便即使在有问题的样本中也能在结果图像中实现最佳分辨率。可以以不同的方式进行这种优化，例如通过确定至少一个照明参数或记录参数的最佳调整或通过脉冲照明使得从荧光染料的三重态到更高三重态的激发或通过用耗尽光照射样品来减少荧光染料的三线态，从而减少漂白。

8. 发明申请

US20110226965A1 INCREASED RESOLUTION MICROSCOPY 有权
标题翻译：增加分辨率显微镜
公开(公告)号：US20110226965A1
公开(公告)日：2011-09-22
申请号：US13131801
申请日：2009-11-14
申请人： Ralf Wolleschensky , Ingo Kleppe , Gerhard Krampert , Michael Kempe
发明人： Ralf Wolleschensky , Ingo Kleppe , Gerhard Krampert , Michael Kempe
IPC分类号： G01N21/64
CPC分类号： G02B21/16 , G01N21/6458 , G02B21/361 , G02B21/367 , G02B27/58
摘要： Method for spatially high-resolution luminescence microscopy in which label molecules in a sample are activated to emit luminescence radiation comprising activating only a subset of the label molecules in the sample, wherein activated label molecules have a distance to the closest activated molecules that is greater or equal to a length which results from a predetermined optical resolution, detecting the luminescence radiation, generating a frame from the luminescence radiation, identifying the geometric locations of the label molecules with a spatial resolution increased above the predetermined optical resolution, repeating the steps and forming a combined image, and controlling the acquisition of the several frames by evaluating at least one of the frames or a group of the frames and modifying at least one variable for subsequent repetitions of the steps of generating frames for combining into an image.
摘要翻译：用于空间高分辨率发光显微镜的方法，其中样品中的标记分子被激活以发射发光，包括仅激活样品中标记分子的一个子集，其中激活的标记分子与最接近的激活分子的距离更大或等于由预定的光学分辨率产生的长度，检测发光辐射，从发光辐射产生帧，以高于预定的光学分辨率的空间分辨率识别标签分子的几何位置，重复步骤并形成并且通过评估所述帧或一组帧中的至少一个并且修改至少一个变量来控制所述几帧的获取，以便后续重复生成用于组合成图像的帧的步骤。

9. 发明授权

US09404867B2 Luminescence microscopy 有权
标题翻译：发光显微镜
公开(公告)号：US09404867B2
公开(公告)日：2016-08-02
申请号：US13518115
申请日：2010-10-22
申请人： Michael Kempe , Ralf Netz , Gerhard Krampert
发明人： Michael Kempe , Ralf Netz , Gerhard Krampert
IPC分类号： G01N21/64 , G02B21/16
CPC分类号： G01N21/6458 , G02B21/16
摘要： A luminescence microscopy method includes a sample being used, which comprises a certain substance, wherein the certain substance can be converted repeatedly from a first state, in which it can be excited into emitting luminescence radiation, into a second state, in which it cannot be excited into emitting luminescence radiation. The substance present in the sample can be brought into the first state by irradiating switch radiation. The certain substance can be excited into emitting luminescence radiation by irradiating excitation radiation. The sample emitting luminescence radiation can be displayed. A high-resolution selection of sample regions extending perpendicularly to a sample surface is carried out by irradiating either the switch radiation or the excitation radiation as structured illumination of the sample. A high-resolution selection of the sample surface is carried out by irradiating the switch radiation and/or the excitation radiation as TIRF illumination of the sample.
摘要翻译：发光显微镜方法包括使用的样品，其包含某种物质，其中某些物质可以从其可被激发发射发光辐射的第一状态重复地转换成第二状态，其中不能是激发发射发光辐射。存在于样品中的物质可以通过照射开关辐射而进入第一状态。可以通过照射激发辐射来激发某些物质发射发光辐射。可以显示发射发光辐射的样品。垂直于样品表面延伸的样品区域的高分辨率选择是通过照射样品的结构照明中的开关辐射或激发辐射来进行的。通过将开关辐射和/或激发辐射照射作为样品的TIRF照明来进行样品表面的高分辨率选择。

10. 发明申请

US20120319007A1 LUMINESCENCE MICROSCOPY 有权
标题翻译：荧光显微镜
公开(公告)号：US20120319007A1
公开(公告)日：2012-12-20
申请号：US13518115
申请日：2010-10-22
申请人： Michael Kempe , Ralf Netz , Gerhard Krampert
发明人： Michael Kempe , Ralf Netz , Gerhard Krampert
IPC分类号： G01N21/64
CPC分类号： G01N21/6458 , G02B21/16
摘要： A luminescence microscopy method includes a sample being used, which comprises a certain substance, wherein the certain substance can be converted repeatedly from a first state, in which it can be excited into emitting luminescence radiation, into a second state, in which it cannot be excited into emitting luminescence radiation. The substance present in the sample can be brought into the first state by irradiating switch radiation. The certain substance can be excited into emitting luminescence radiation by irradiating excitation radiation. The sample emitting luminescence radiation can be displayed. A high-resolution selection of sample regions extending perpendicularly to a sample surface is carried out by irradiating either the switch radiation or the excitation radiation as structured illumination of the sample. A high-resolution selection of the sample surface is carried out by irradiating the switch radiation and/or the excitation radiation as TIRF illumination of the sample.
摘要翻译：发光显微镜方法包括使用的样品，其包含某种物质，其中某些物质可以从其可被激发发射发光辐射的第一状态重复地转换到第二状态，其中不能是激发发射发光辐射。存在于样品中的物质可以通过照射开关辐射而进入第一状态。可以通过照射激发辐射来激发某些物质发射发光辐射。可以显示发射发光辐射的样品。垂直于样品表面延伸的样品区域的高分辨率选择是通过照射样品的结构照明中的开关辐射或激发辐射来进行的。通过将开关辐射和/或激发辐射照射作为样品的TIRF照明来进行样品表面的高分辨率选择。

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