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71. 发明申请

US20130020473A1 MICROSCOPE AND METHOD FOR DETECTING SAMPLE LIGHT 有权
标题翻译：显微镜和检测样品光的方法
公开(公告)号：US20130020473A1
公开(公告)日：2013-01-24
申请号：US13637122
申请日：2011-03-17
申请人： Thomas Kalkbrenner , Ralf Wolleschensky
发明人： Thomas Kalkbrenner , Ralf Wolleschensky
IPC分类号： G02F1/01 , G02B21/00
CPC分类号： G02B21/0056 , G02B21/0032 , G02B21/0076 , G02B26/06
摘要： Microscope and method for detecting sample light, having at least one illuminating beam which is partially phase-modulated with a modulation frequency along the cross section thereof and a microscope objective for intensity-modulated focusing of the illuminating beam into a sample. The microscope has a detection beam path that has at least one demodulator. At least one electro-optical modulator (EOM) is used for phase modulation of at least a part, preferably half, of the illuminating beam, or different portions or halves of the illuminating beam are modulated differently, preferably anti-phase, by anti-phase control of piezoelectric elements, or acousto-optical modulators for splitting into a plurality of partial beam paths. Optic elements are provided for partial phase modulation of the excitation beam. Actuating elements are provided for setting the phase difference, or at least one optic modulator, preferably an acousto-optical modulator (AOM) is used for demodulation in the detection, or a change in the mode of operation of the detectors for demodulation takes place. In the case of foci distribution, produced for example by a spinning micro-lens disc or multi-spot generation, the individual foci are subjected to an intensity modulation, either by arranging a half-space phase mask in a pupil plane of the objective or by individually manipulating each partial beam by means of the partial phase modulation of said partial beam with a modulation frequency along the cross section thereof or by manipulating a beam by means of the partial phase modulation of said beam with a modulation frequency along the cross section thereof and subsequent subdivision of said beam.
摘要翻译：用于检测样品光的显微镜和方法，具有至少一个沿其横截面以调制频率部分相位调制的照明光束和用于将照明光束强度调制到样本中的显微镜物镜。显微镜具有至少一个解调器的检测光束路径。至少一个电光调制器（EOM）被用于照明光束的至少一部分，优选一半的相位调制，或者照明光束的不同部分或一半被不同地调制，优选是反相的，压电元件的相位控制或用于分离成多个部分光束路径的声光调制器。提供光学元件用于激发光束的部分相位调制。提供用于设置相位差的驱动元件，或者至少一个光调制器，优选地，声光调制器（AOM）用于检测中的解调，或者发生用于解调的检测器的操作模式的变化。在例如通过旋转微透镜盘或多点产生产生的焦点分布的情况下，通过在物镜的瞳面中布置半空间相位掩模来对各个焦点进行强度调制，或通过利用沿着其横截面的调制频率的所述部分波束的部分相位调制来单独操纵每个部分波束，或者通过沿着其横截面的调制频率通过所述波束的部分相位调制来操纵波束并且随后细分所述光束。

72. 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。

73. 发明申请

US20110284767A1 COMBINATION MICROSCOPY 有权
标题翻译：组合显微镜
公开(公告)号：US20110284767A1
公开(公告)日：2011-11-24
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，并且生成第三显微镜图像 STED，ESA或RESOLFT，其中所获得的图像被叠加。

74. 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。

75. 发明授权

US07999238B2 Arrangement for processing signals at the output of a multichannel detector 有权
标题翻译：用于在多通道检测器的输出端处理信号的布置
公开(公告)号：US07999238B2
公开(公告)日：2011-08-16
申请号：US12375389
申请日：2007-07-16
申请人： Ralf Wolleschensky , Mirko Liedtke
发明人： Ralf Wolleschensky , Mirko Liedtke
IPC分类号： G01N21/64
CPC分类号： G02B21/0084 , G01N21/6408 , G02B21/008
摘要： An arrangement for signal processing at the output of a multichannel detector in the spectrally resolved acquisition of time-variable fluorescence phenomena in a microscope, particularly lifetime measurements, is characterized in that an FPGA (free programmable gate array) is arranged downstream of the output of the multichannel detector.
摘要翻译：在显微镜中对时变荧光现象进行光谱分辨采集，特别是寿命测量的多通道检测器的输出处的信号处理装置的特征在于，FPGA（自由可编程门阵列）布置在多通道检测器。

76. 发明授权

US07852474B2 Spectral analysis unit with a diffraction grating 有权
$Spectral analysis unit with a diffraction grating$
标题翻译：具有衍射光栅的光谱分析单元
公开(公告)号：US07852474B2
公开(公告)日：2010-12-14
申请号：US11785153
申请日：2007-04-16
申请人： Hans-Juergen Dobschal , Ralf Wolleschensky , Wolfgang Bathe , Joerg Steinert
发明人： Hans-Juergen Dobschal , Ralf Wolleschensky , Wolfgang Bathe , Joerg Steinert
IPC分类号： G01J3/28
CPC分类号： G02B21/0064 , G01J3/02 , G01J3/0208 , G01J3/021 , G01J3/0259 , G01J3/1804 , G02B21/16
摘要： A spectral analytical unit for acting on a parallel light bundle having different wavelengths. The spectral analytical unit includes a diffraction grating on which the light bundle falls, the diffraction grating splitting the different wavelengths through diffraction in first spectral directions defining a light bundle diffraction order 1 without recycle, and the diffraction grating bending the light bundle in second directions defining a light bundle diffraction order 0 without recycle, a detector line made up of a plurality of elements, optics for focusing the split light bundle diffraction order 1 without recycle on the detector line, evaluation electronics connected to the detector line for obtaining data related to a created spectrum, and a deflecting device wherein the diffraction order 0 light bundle without recycle meets on the deflecting device which is so directed and positioned that this light bundle falls on the diffraction grating thereby creating a reflected diffraction order 1 light bundle with first recycle and a reflected diffraction order 0 light bundle with first recycle whereby the diffraction order 1 without recycle and the reflected diffraction order 1 light bundle with first recycle each of a part wavelength range are impressed through the optics on a single element of the detector line.
摘要翻译：用于作用于具有不同波长的平行光束的光谱分析单元。光谱分析单元包括光束落在其上的衍射光栅，衍射光栅通过在限定光束衍射级数1而不进行再循环的第一光谱方向上通过衍射分离不同波长，并且衍射光栅在限定第二方向的第二方向上折射光束没有再循环的光束衍射级0，由多个元件组成的检测器线，用于将分束光束衍射级1聚焦在检测器线路上而不再循环的光学器件，连接到检测器线的评估电子设备，用于获得与以及偏转装置，其中没有再循环的衍射级0光束在偏转装置上相遇，所述偏转装置被定向和定位成使得该光束落在衍射光栅上，从而产生具有第一再循环的反射衍射级1光束和反射衍射级0光束与fi 第一次循环，其中没有再循环的衍射级1和具有部分波长范围的第一次再循环的反射衍射级1光束通过检测器线的单个元件上的光学器件施加。

77. 发明授权

US07688504B2 Optical scanning microscope 有权
标题翻译：光学扫描显微镜
公开(公告)号：US07688504B2
公开(公告)日：2010-03-30
申请号：US12320331
申请日：2009-01-23
申请人： Ralf Wolleschensky
发明人： Ralf Wolleschensky
IPC分类号： G02B21/06
CPC分类号： G02B21/0032 , G02B21/002
摘要： Microscope, in particular an optical scanning microscope with illumination of a specimen via a beam splitter, which is arranged in an objective pupil and includes at least a reflecting first portion and at least a transmitting second portion, whereby the reflecting portion serves to couple in the illumination light and the transmitting portion serves to pass the detection light in the detection direction or the transmitting portion serves to couple in the illumination light and the reflecting portion serves to couple out the detection light, with a first scanning arrangement. Means are provided in the detection light path for the overlay of at least one further scanning arrangement for illumination and detection.
摘要翻译：显微镜，特别是具有通过分束器照射样品的光学扫描显微镜，其被布置在物镜光瞳中，并且至少包括反射第一部分和至少透射第二部分，由此反射部分用于在照明光和透射部分用于使检测方向上的检测光通过，或者透射部分用于在照明光中耦合，并且反射部分用于以第一扫描装置耦合检测光。在用于照明和检测的至少一个另外的扫描装置的覆盖的检测光路中提供了装置。

78. 发明申请

US20090250627A1 ARRANGEMENT FOR PROCESSING SIGNALS AT THE OUTPUT OF A MULTICHANNEL DETECTOR 有权
标题翻译：在多通道检测器的输出处理信号的布置
公开(公告)号：US20090250627A1
公开(公告)日：2009-10-08
申请号：US12375389
申请日：2007-07-16
申请人： Ralf Wolleschensky , Mirko Liedtke
发明人： Ralf Wolleschensky , Mirko Liedtke
IPC分类号： G01N21/64
CPC分类号： G02B21/0084 , G01N21/6408 , G02B21/008
摘要： An arrangement for signal processing at the output of a multichannel detector in the spectrally resolved acquisition of time-variable fluorescence phenomena in a microscope, particularly lifetime measurements, is characterized in that an FPGA (free programmable gate array) is arranged downstream of the output of the multichannel detector.
摘要翻译：在显微镜中对时变荧光现象进行光谱分辨采集，特别是寿命测量的多通道检测器的输出处的信号处理装置的特征在于，FPGA（自由可编程门阵列）布置在多通道检测器。

79. 发明申请

US20090161207A1 Optical scanning microscope 有权
标题翻译：光学扫描显微镜
公开(公告)号：US20090161207A1
公开(公告)日：2009-06-25
申请号：US12320331
申请日：2009-01-23
申请人： Ralf Wolleschensky
发明人： Ralf Wolleschensky
IPC分类号： G02B21/06
CPC分类号： G02B21/0032 , G02B21/002
摘要： Microscope, in particular an optical scanning microscope with illumination of a specimen via a beam splitter, which is arranged in an objective pupil and includes at least a reflecting first portion and at least a transmitting second portion, whereby the reflecting portion serves to couple in the illumination light and the transmitting portion serves to pass the detection light in the detection direction or the transmitting portion serves to couple in the illumination light and the reflecting portion serves to couple out the detection light, with a first scanning arrangement. Means are provided in the detection light path for the overlay of at least one further scanning arrangement for illumination and detection.
摘要翻译：显微镜，特别是具有通过分束器照射样品的光学扫描显微镜，其被布置在物镜光瞳中，并且至少包括反射第一部分和至少透射第二部分，由此反射部分用于在照明光和透射部分用于使检测方向上的检测光通过，或者透射部分用于在照明光中耦合，并且反射部分用于以第一扫描装置耦合检测光。在用于照明和检测的至少一个另外的扫描装置的覆盖的检测光路中提供了装置。

80. 发明授权

US07485875B2 Resolution-enhanced luminescence microscopy 有权
标题翻译：分辨率增强发光显微镜
公开(公告)号：US07485875B2
公开(公告)日：2009-02-03
申请号：US11491781
申请日：2006-07-24
申请人： Ralf Wolleschensky , Michael Kempe
发明人： Ralf Wolleschensky , Michael Kempe
IPC分类号： G21H3/02 , G01N21/64
CPC分类号： G01N21/6458 , G02B21/0072 , G02B21/0076 , G02B21/16
摘要： A resolution-enhanced luminescence microscopy method, wherein a sample is excited so as to luminesce, and thus to emit a given luminescence radiation, by irradiation of exciting radiation and an image of the luminescent sample is obtained, wherein the luminescent sample is transferable from a first state of luminescence, in which first state the sample's excitability for emission of the given luminescence radiation increases up to a maximum value as the exciting radiation power increases, into a second state of luminescence, in which second state the sample has reduced excitability for emission of the given luminescence radiation relative to the first state, wherein the maximum value is assigned to a threshold value of exciting radiation power and the sample is transferable into the second state by irradiation of exciting radiation power above the threshold value, the sample being brought into the first state in partial areas and being brought into the second state in adjacent partial areas by irradiating exciting radiation with an exciting radiation distribution having at least one spatial power maximum above the threshold value, the image of the luminescent sample comprising sample areas being in the first state and sample areas being in the second state, sample areas being in the first state contributing predominantly to the image and the image thus having an enhanced spatial resolution with respect to the exciting radiation distribution.
摘要翻译：一种分辨率增强型发光显微镜方法，其中通过照射激发辐射激发样品以发光，从而发射给定的发光辐射，并且获得发光样品的图像，其中发光样品可从发光的第一状态，其中第一状态是给定发光辐射的发射的兴奋性随着激发辐射功率的增加而增加到最大值，变成第二种状态，其中第二种状态是样品具有降低的发射兴奋性的给定发光辐射相对于第一状态，其中最大值被分配给激发辐射功率的阈值，并且样品可通过超过阈值的激发辐射功率的照射而转移到第二状态，将样品置入部分地区的第一个状态，并在相邻的部分地区进入第二个状态通过用具有高于阈值的至少一个空间功率最大值的激发辐射分布来照射激发辐射的区域，所述发光样品的图像包括样品区域处于第一状态，样品区域处于第二状态，样品区域在主要对图像和图像贡献的第一状态因此相对于激发辐射分布具有增强的空间分辨率。

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