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1. 发明授权

US08735444B2 Fluorinated rhodamines as photostable fluorescent dyes for labelling and imaging techniques 有权
标题翻译：氟化罗丹明作为用于标记和成像技术的光稳定荧光染料
公开(公告)号：US08735444B2
公开(公告)日：2014-05-27
申请号：US13380148
申请日：2009-06-26
申请人： Stefan W. Hell , Vladimir N. Belov , Gyuzel Mitronova , Mariano Bossi , Gael Moneron , Christian A. Wurm , Stefan Jakobs , Christian Eggeling , Jakob Bierwagen , Lars Meyer
发明人： Stefan W. Hell , Vladimir N. Belov , Gyuzel Mitronova , Mariano Bossi , Gael Moneron , Christian A. Wurm , Stefan Jakobs , Christian Eggeling , Jakob Bierwagen , Lars Meyer
IPC分类号： A61K31/352 , C07D311/82
CPC分类号： C07D311/82 , A61K49/0041 , A61K49/0058 , C07K1/13 , C09B11/24 , G01N33/582
摘要： The present invention relates to novel fluorinated 3,6-diaminoxanthene compounds derived from the basic structural formula (I) and to their uses as photostable fluorescent dyes, e.g. for immunostainings and spectroscopic and microscopic applications, in particular in conventional microscopy, stimulated emission depletion (STED) reversible saturable optically linear fluorescent transitions (RESOLFT) microscopy, and fluorescence correlation spectroscopy. The claimed compounds are also useful as molecular probes in various spectroscopic applications.
摘要翻译：本发明涉及衍生自基本结构式（I）的新型氟化3,6-二氨基呫吨化合物及其作为光稳定荧光染料的用途。用于免疫染色和光谱和微观应用，特别是在常规显微镜，受激发射耗尽（STED）可逆饱和光学线性荧光转换（RESOLFT）显微镜和荧光相关光谱学中。所要求保护的化合物也可用作各种光谱应用中的分子探针。

2. 发明授权

US08617827B2 Photoactivable fluorescent dyes for optical microscopy and imaging techniques 有权
标题翻译：用于光学显微镜和成像技术的可光化荧光染料
公开(公告)号：US08617827B2
公开(公告)日：2013-12-31
申请号：US13395323
申请日：2009-09-10
申请人： Stefan W. Hell , Vladimir N. Belov , Vadim P. Boyarskiy , Christian A. Wurm , Stefan Jakobs , Claudia Geisler
发明人： Stefan W. Hell , Vladimir N. Belov , Vadim P. Boyarskiy , Christian A. Wurm , Stefan Jakobs , Claudia Geisler
IPC分类号： G01N33/53 , C07D285/06 , C12Q1/02 , C09B46/00
CPC分类号： C09B11/24
摘要： The present invention relates to novel photoactivable rhodamine or carbopyronine derivatives of the following general formulae G1-G4 (I), G1: A1=O, A2=N, A3=C; G2: A1=S, A2=N, A3=C; G3: A1=O, A2=O, A3=N; G4: A1=S, A2=O, A3=N; comprising UV light absorbing chromophores which after photolysis by irradiation at 254-490 nm (preferably at 375-420 nm) generate fluorescent rhodamine or carbopyronine derivatives and small non-toxic fragments such as N2 or N2O. The invention also relates to methods for preparing such compounds and to the use of these compounds for optical microscopy and imaging techniques.
摘要翻译：本发明涉及以下通式G1-G4（I），G1：A1 = O，A2 = N，A3 = C的新型可光致发生罗丹明或卡吡龙衍生物。 G2：A1 = S，A2 = N，A3 = C; G3：A1 = O，A2 = O，A3 = N; G4：A1 = S，A2 = O，A3 = N; 包括紫外光吸收发色团，其在254-490nm（优选在375-420nm）下照射光解后产生荧光罗丹明或卡吡喃衍生物和小的无毒片段如N 2或N 2 O。本发明还涉及制备这些化合物的方法以及这些化合物用于光学显微镜和成像技术的用途。

3. 发明授权

US08039815B2 Fluorescent light microscope for measuring a sample using red-shifted stokes lines 有权
标题翻译：荧光光学显微镜用于使用红移斯托克斯线测量样品
公开(公告)号：US08039815B2
公开(公告)日：2011-10-18
申请号：US12753349
申请日：2010-04-02
申请人： Stefan W. Hell , Brian Rankin , Robert Kellner , Jaydev Jethwa , Thorsten Staudt
发明人： Stefan W. Hell , Brian Rankin , Robert Kellner , Jaydev Jethwa , Thorsten Staudt
IPC分类号： G01N21/64
CPC分类号： G02B21/16 , G01N21/6458 , G01N21/65 , G01N2021/655 , H01S3/302
摘要： A fluorescent light microscope for measuring a sample comprises a light source providing transfer light having a transfer wavelength for transferring a fluorescent dye in the sample from one state into another state, and a detector which measures fluorescent light from the sample with spatial resolution. The light source comprises a laser, an optical wave guide connected to the laser, and a wavelength-selective device connected to the optical wave guide. The laser emits pump light of a pump wavelength other than the transfer wavelength and injects the pump light into the optical wave guide. The pump light, due to Raman scattering being stimulated in the optical wave guide, generates a light spectrum emerging from the optical wave guide which has, besides the pump wavelength, at least one red-shifted Stokes line whose full width at half maximum is smaller than half of its distance to its next neighbor line on the blue side of the spectrum; and the wavelength-selective device singles out the transfer light by its transfer wavelength from the red-shifted Stokes lines of the light spectrum.
摘要翻译：用于测量样品的荧光显微镜包括提供具有用于将样品中的荧光染料从一种状态转移到另一状态的转印波长的转移光，以及以空间分辨率测量来自样品的荧光的检测器。光源包括激光器，连接到激光器的光波导和连接到光波导的波长选择器件。激光器发射除了转移波长之外的泵波长的泵浦光，并将泵浦光注入到光波导中。由于在光波导中被拉曼散射的泵浦光产生从光波导出射的光谱，除了泵浦波长之外，还有至少一个红移斯托克斯线，其半峰全宽较小超过其在光谱蓝色边上的下一个相邻线的距离的一半; 并且波长选择装置通过其来自光谱的红移斯托克斯线的转移波长来分离转移光。

4. 发明授权

US07863585B2 STED-fluorescent light microscopy with two-photon excitation 有权
标题翻译：具有双光子激发的STED-荧光光学显微镜
公开(公告)号：US07863585B2
公开(公告)日：2011-01-04
申请号：US12706388
申请日：2010-02-16
申请人： Stefan W. Hell , Katrin Willig
发明人： Stefan W. Hell , Katrin Willig
IPC分类号： G01N21/64
CPC分类号： G01N21/6458 , G01N21/636 , G02B21/0076
摘要： A method of high spatial resolution imaging a structure in a sample comprises: marking the structure with molecules of a fluorescent dye; selecting a first wavelength for excitation light which excites the molecules of the fluorescent dye via a multi photon process for spontaneous emission of fluorescent light; focusing pulses of the excitation light into the sample to excite those molecules of the fluorescent dye present in a focal area of the focused excitation light; selecting a second wavelength shorter than the first wavelength for de-excitation light which de-excites excited molecules of the fluorescent dye prior to their spontaneous emission; during a plurality of the pulses of the excitation light, continuously directing the de-excitation light onto the sample to de-excite excited molecules of the fluorescent dye, which are located outside an measurement area which is a fraction of the focal area; and recording the fluorescent light spontaneously emitted by the molecules of the fluorescent dye in the sample.
摘要翻译：对样品中的结构进行高空间分辨率成像的方法包括：用荧光染料的分子标记结构; 选择激发光的第一波长，其通过用于自发发射荧光的多光子过程激发荧光染料的分子; 将激发光的脉冲聚焦到样品中以激发存在于聚焦激发光的焦点区域中的荧光染料的那些分子; 选择比第一波长短的第二波长用于去激发荧光染料在其自发发射之前激发分子的去激发光; 在激发光的多个脉冲期间，将去激发光连续地引导到样品上以去激发位于焦点区域的一部分的测量区域外的荧光染料的激发分子; 并记录由荧光染料分子自发发射的荧光。

5. 发明申请

US20120212750A1 Three-Dimensionally Localizing Light Emitting Molecules of Unknown Orientation and Unknown Z-Position 有权
标题翻译：未知定位和未知Z位置的三维定位发光分子
公开(公告)号：US20120212750A1
公开(公告)日：2012-08-23
申请号：US13441043
申请日：2012-04-06
申请人： Johann Engelhardt , Stefan W. Hell , Jan Keller-Findeisen
发明人： Johann Engelhardt , Stefan W. Hell , Jan Keller-Findeisen
IPC分类号： G01B11/14
CPC分类号： G02B21/0076 , G01N21/6456 , G01N2021/6463
摘要： To the end of three-dimensionally localizing light emitting marker entities of unknown orientation and unknown position in a sample, the light emitted by each single marker entity is imaged in at least two different ways onto at least one detection plane which corresponds to a focal plane (13) in the sample resulting in at least two images of the marker entity. Virtual x- and y-positions of the marker entity in parallel to the focal plane (13) are separately determined from the emitted light intensity distribution over each image of the marker entity. Further, the z-position of the marker entity normal to the focal plane is determined from the emitted light intensity distributions over the images of the marker entity. The real x- and y-positions of the marker entity in parallel to the focal plane (13) are determined based on its virtual x- and y-positions and on its z-position.
摘要翻译：在三维定位样品中未知取向和未知位置的发光标记实体的三维定位结束时，由每个单个标记实体发射的光以至少两种不同的方式成像到至少一个对应于焦平面的检测平面（13），导致标记实体的至少两个图像。标记实体与焦平面（13）平行的虚拟x和y位置根据标记实体的每个图像上的发射光强度分布分别确定。此外，从标记实体的图像上的发射光强度分布确定垂直于焦平面的标记实体的z位置。标记实体与焦平面（13）平行的真实x和y位置基于其虚拟x和y位置及其z位置来确定。

6. 发明申请

US20100140506A1 METHOD OF DETERMINING A MEASUREMENT VALUE ON THE BASIS OF SINGLE MOLECULE EVENTS 审中-公开
标题翻译：基于单分子事件确定测量值的方法
公开(公告)号：US20100140506A1
公开(公告)日：2010-06-10
申请号：US12687969
申请日：2010-01-15
申请人： Christian EGGELING , Andreas SCHÖNLE , Stefan W. HELL , Michael HILBERT
发明人： Christian EGGELING , Andreas SCHÖNLE , Stefan W. HELL , Michael HILBERT
IPC分类号： G01N21/64
CPC分类号： G01N21/6428 , G01N21/6408 , G01N21/6458 , G01N2021/6419 , G01N2021/6439
摘要： A method of determining a measurement value on the basis of a plurality of single molecule events of marker molecules in a sample comprises the steps of selecting the marker molecules from a group of marker molecules which are transferable between a measurable state in which a measurement signal necessary for determining the at least one measurement value is obtainable from the marker molecules and a non-measurable state, of providing the marker molecules in the sample at such an absolute concentration that the at least one measurement value is not determinable, if all marker molecules are in their measurable state, and adjusting a measurement concentration of the marker molecules in the measurable state by means of applying the physical signal to the sample at such an intensity that the at least one measurement value is determinable within a defined measurement area of the sample.
摘要翻译：基于样品中标记分子的多个单分子事件确定测量值的方法包括以下步骤：从一组标记分子中选择标记分子，所述标记分子可在可测量状态之间转移，其中测量信号必需用于确定所述至少一个测量值可从所述标记分子和不可测量的状态获得，以如下绝对浓度提供所述标记分子，所述绝对浓度至少一个测量值不能确定，如果所有标记分子是在可测量的状态下，并且以可测量状态的标记分子的测量浓度通过将样品的物理信号以样品的限定测量区域内的至少一个测量值确定的强度来调节。

7. 发明申请

US20140220588A1 FLUORESCENT DYES WITH PHOSPHORYLATED HYDROXYMETHYL GROUPS AND THEIR USE IN LIGHT MICROSCOPY AND IMAGING TECHNIQUES 有权
标题翻译：荧光染料与羟基甲基纤维素组合及其在光学显微镜和成像技术中的应用
公开(公告)号：US20140220588A1
公开(公告)日：2014-08-07
申请号：US13702252
申请日：2011-10-18
申请人： Stefan W. Hell , Shamil Nizamov , Gerald Donnert , Kirill Kolmakov , Heiko Schill , Lars Kastrup , Christian A. Wurm , Vladimir N. Belov , Johanna Bueckers , Katrin Willig
发明人： Stefan W. Hell , Shamil Nizamov , Gerald Donnert , Kirill Kolmakov , Heiko Schill , Lars Kastrup , Christian A. Wurm , Vladimir N. Belov , Johanna Bueckers , Katrin Willig
IPC分类号： G01N33/52
CPC分类号： G01N33/52 , C09B11/24 , C09B57/02 , C09B57/10 , G01N33/533 , G01N33/582 , G01N2458/00
摘要： The invention relates to novel fluorescent dyes with phosphorylated hydroxymethyl groups, a method for preparing the same as well as to their use in imaging techniques. The fluorescent dyes are coumarin, rhodamine or BODIPY dyes having of one of the following general formulae I-III: wherein W=OP(O) Y1Y2 or P(O) Y1Y2, where Y1 and Y2 independently denote any of the following residues: OH, O(−), ORa and ORb, NHRa and NHRb, NRaRb and NRcRd, ORa and NHRb, ORa and NRbRc, NHRa and NRbRc; and any salt thereof.
摘要翻译：本发明涉及具有磷酸化羟甲基的新型荧光染料，其制备方法以及其在成像技术中的应用。荧光染料是具有以下通式I-III之一的香豆素，罗丹明或BODIPY染料：其中W = OP（O）Y1Y2或P（O）Y1Y2，其中Y1和Y2独立地表示以下任何残基：OH ，O（ - ），ORa和ORb，NHRa和NHRb，NRaRb和NRcRd，ORa和NHRb，ORa和NRbRc，NHRa和NRbRc; 及其任何盐。

8. 发明申请

US20110081653A1 High Spatial Resolution Imaging of a Structure of Interest in a Specimen 有权
标题翻译：样本中兴趣结构的高空间分辨率成像
公开(公告)号：US20110081653A1
公开(公告)日：2011-04-07
申请号：US12949159
申请日：2010-11-18
申请人： Stefan W. HELL , Jonas FÖLLING , Christian EGGELING , Alexander EGNER , Andreas SCHÖNLE , Mariano BOSSI
发明人： Stefan W. HELL , Jonas FÖLLING , Christian EGGELING , Alexander EGNER , Andreas SCHÖNLE , Mariano BOSSI
IPC分类号： G01N21/64 , G21K5/00 , C12Q1/68
CPC分类号： G01N21/6458 , G01N21/6428 , G02B21/16 , G02B21/367 , G02B27/58
摘要： In high spatial resolution imaging, a structure in a specimen is marked with a substance which, in a first electronic state, is excited by light of one wavelength to emit fluorescent light, which is also converted from its first into a second electronic state by that light, and which returns from its second into its first electronic state. The specimen is imaged onto a sensor at a spatial resolution not resolving an average spacing between neighboring molecules of the substance, and exposed to the light at such an intensity that the molecules in the first state are alternately excited to emit fluorescent light and converted into their second state, and that at least 10% of the molecules presently in their first state lie at a distance from their closest neighboring molecules in their first state which is greater than the spatial resolution of the imaging onto the sensor.
摘要翻译：在高空间分辨率成像中，样本中的结构用物质标记，该物质在第一电子状态下被一个波长的光激发以发射荧光，其也从其第一个电子状态转换成第二电子状态光，从第二个回到第一个电子状态。将样本以不分辨物质的相邻分子之间的平均间隔的空间分辨率成像到传感器上，并以如此强度暴露于光，使得第一状态下的分子被交替地激发以发射荧光并转换成它们并且目前处于其第一状态的至少10％的分子处于与它们的第一状态中最接近的相邻分子相距一定距离，其大于成像在传感器上的空间分辨率。

9. 发明申请

US20100187438A1 Fluorescent Light Microscope For Measuring a Sample Using Red-Shifted Stokes Lines 有权
标题翻译：荧光灯显微镜用于测量样品使用红移斯托克斯线
公开(公告)号：US20100187438A1
公开(公告)日：2010-07-29
申请号：US12753349
申请日：2010-04-02
申请人： Stefan W. Hell , Brian RANKIN , Robert KELLNER , Jaydev JETHWA , Thorsten STAUDT
发明人： Stefan W. Hell , Brian RANKIN , Robert KELLNER , Jaydev JETHWA , Thorsten STAUDT
IPC分类号： G01N21/64 , G01J1/58
CPC分类号： G02B21/16 , G01N21/6458 , G01N21/65 , G01N2021/655 , H01S3/302
摘要： A fluorescent light microscope for measuring a sample comprises a light source providing transfer light having a transfer wavelength for transferring a fluorescent dye in the sample from one state into another state, and a detector which measures fluorescent light from the sample with spatial resolution. The light source comprises a laser, an optical wave guide connected to the laser, and a wavelength-selective device connected to the optical wave guide. The laser emits pump light of a pump wavelength other than the transfer wavelength and injects the pump light into the optical wave guide. The pump light, due to Raman scattering being stimulated in the optical wave guide, generates a light spectrum emerging from the optical wave guide which has, besides the pump wavelength, at least one red-shifted Stokes line whose full width at half maximum is smaller than half of its distance to its next neighbor line on the blue side of the spectrum; and the wavelength-selective device singles out the transfer light by its transfer wavelength from the red-shifted Stokes lines of the light spectrum.
摘要翻译：用于测量样品的荧光显微镜包括提供具有用于将样品中的荧光染料从一种状态转移到另一状态的转印波长的转移光，以及以空间分辨率测量来自样品的荧光的检测器。光源包括激光器，连接到激光器的光波导和连接到光波导的波长选择器件。激光器发射除了转移波长之外的泵波长的泵浦光，并将泵浦光注入到光波导中。由于在光波导中被拉曼散射的泵浦光产生从光波导出射的光谱，除了泵浦波长之外，还有至少一个红移斯托克斯线，其半峰全宽较小超过其在光谱蓝色边上的下一个相邻线的距离的一半; 并且波长选择装置通过其来自光谱的红移斯托克斯线的转移波长来分离转移光。

10. 发明授权

US06891670B2 Double confocal scanning microscope 有权
标题翻译：双共焦扫描显微镜
公开(公告)号：US06891670B2
公开(公告)日：2005-05-10
申请号：US09683713
申请日：2002-02-06
申请人： Hilmar Gugel , Joerg Bewersdorf , Stefan W. Hell
发明人： Hilmar Gugel , Joerg Bewersdorf , Stefan W. Hell
IPC分类号： G02B21/00 , G02B21/14 , G02B21/06
CPC分类号： G02B21/0032 , G02B21/0056 , G02B21/0068 , G02B21/0076
摘要： The present invention concerns a double confocal scanning microscope having an illuminating beam path (1) of a light source (2) and a detection beam path (3) of a detector (4), and in order to eliminate at their cause the problems of reconstruction methods. To do so, at least one optical component (24, 25) acting on the illuminating and/or detection beam path (1, 3) is provided, and is configured in such a way that it influences the amplitude and/or phase and/or polarization of the light; and the characteristics of the double confocal illumination and/or detection are thereby modifiable.
摘要翻译：本发明涉及一种具有光源（2）的照明光束路径（1）和检测器（4）的检测光束路径（3）的双共焦扫描显微镜，并且为了消除其原因，重建方法。为此，提供作用在照明和/或检测光束路径（1,3）上的至少一个光学部件（24,25），并且被配置成使得其影响幅度和/或相位和/ 或光的极化; 因此可以修改双共聚焦照明和/或检测的特性。

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