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1. 发明申请

US20120100559A1 NOVEL HYDROPHILIC AND LIPOPHILIC RHODAMINES FOR LABELLING AND IMAGING 有权
标题翻译：用于标签和成像的新型水解和液体罗丹明
公开(公告)号：US20120100559A1
公开(公告)日：2012-04-26
申请号：US13266775
申请日：2010-04-26
申请人： Stefan Hell , Vladimir N. Belov , Kirill Kolmakov , Volker Westphal , Marcel Lauterbach , Stefan Jakobs , Christian Wurm , Christian Eggeling , Christian Ringemann
发明人： Stefan Hell , Vladimir N. Belov , Kirill Kolmakov , Volker Westphal , Marcel Lauterbach , Stefan Jakobs , Christian Wurm , Christian Eggeling , Christian Ringemann
IPC分类号： G01N33/567 , C07D491/22 , C07H21/00 , C12Q1/02 , C07H1/00
CPC分类号： C07D491/22 , C09B11/24 , G01N33/582
摘要： The invention relates to novel and improved photostable rhodamine dyes of the general structural formulae I or II and their uses as fluorescent markers, e.g. for immunostainings and spectroscopic and microscopic applications, in particular in conventional and stimulated emission depletion (STED) microscopy and fluorescence correlation spectroscopy. The partially deuterated analogues are useful as molecular mass distribution tags in mass spectroscopic applications. wherein R1=an unsubstituted or substituted alkyl group, including a cycloalkyl group, or heterocycloalkyl group; R2═H, an unsubstituted or substituted alkyl group, including a cycloalkyl group, or heterocycloalkyl group, or an unsubstituted or substituted aryl group or heteroaryl group, or any combination of such groups; X═CH2, C═O, C═NORa, C═NNRaNRb, CH(ORa), O, S, SO, SO2, or any other derivatives of these groups, with Ra and Rb independently being H or an organic residue, in particular an unsubstituted or substituted (cyclo) alkyl group or heterocycloalkyl group, an unsubstituted or substituted aryl group or heteroaryl group; Z=a negatively charged group with 1, 2, 3, 4 or 5 charges per anion.
摘要翻译：本发明涉及通用结构式I或II的新型和改进的光稳定罗丹明染料及其作为荧光标记物的用途。用于免疫染色和光谱和微观应用，特别是在常规和受激发射耗尽（STED）显微镜和荧光相关光谱学中。部分氘代类似物可用作质谱分析应用中的分子量分布标签。其中R1 =未取代或取代的烷基，包括环烷基或杂环烷基; R 2 = H，未取代或取代的烷基，包括环烷基或杂环烷基，或未取代或取代的芳基或杂芳基，或这些基团的任何组合; X = CH 2，C = O，C = NOR a，C = NR N R a R b，CH（OR a），O，S，SO，SO 2或这些基团的任何其它衍生物，其中Ra和Rb独立地为H或有机残基特别是未取代或取代的（环）烷基或杂环烷基，未取代或取代的芳基或杂芳基; Z =带负电荷的基团，每个阴离子具有1,2,3,4或5个电荷。

2. 发明授权

US08580579B2 Hydrophilic and lipophilic rhodamines for labelling and imaging 有权
标题翻译：亲水和亲脂罗丹明用于标记和成像
公开(公告)号：US08580579B2
公开(公告)日：2013-11-12
申请号：US13266775
申请日：2010-04-26
申请人： Stefan Hell , Vladimir N. Belov , Kirill Kolmakov , Volker Westphal , Marcel Lauterbach , Stefan Jakobs , Christian Wurm , Christian Eggeling , Christian Ringemann
发明人： Stefan Hell , Vladimir N. Belov , Kirill Kolmakov , Volker Westphal , Marcel Lauterbach , Stefan Jakobs , Christian Wurm , Christian Eggeling , Christian Ringemann
IPC分类号： G01N33/533 , G01N33/532 , G01N21/76
CPC分类号： C07D491/22 , C09B11/24 , G01N33/582
摘要： The invention relates to novel and improved photostable rhodamine dyes of the general structural formulae I or II and their uses as fluorescent markers, e.g. for immunostainings and spectroscopic and microscopic applications, in particular in conventional and stimulated emission depletion (STED) microscopy and fluorescence correlation spectroscopy. The partially deuterated analogues are useful as molecular mass distribution tags in mass spectroscopic applications, wherein R1=an unsubstituted or substituted alkyl group, including a cycloalkyl group, or heterocycloalkyl group; R2=H, an unsubstituted or substituted alkyl group, including a cycloalkyl group, or heterocycloalkyl group, or an unsubstituted or substituted aryl group or heteroaryl group, or any combination of such groups; X=CH2, C═O, C═NORa, C═NNRaNRb, CH(ORa), O, S, SO, SO2, or any other derivatives of these groups, with Ra and Rb independently being H or an organic residue, in particular an unsubstituted or substituted (cyclo)alkyl group or heterocycloalkyl group, an unsubstituted or substituted aryl group or heteroaryl group; Z=a negatively charged group with 1, 2, 3, 4 or 5 charges per anion.
摘要翻译：本发明涉及通用结构式I或II的新型和改进的光稳定罗丹明染料及其作为荧光标记物的用途。用于免疫染色和光谱和微观应用，特别是在常规和受激发射耗尽（STED）显微镜和荧光相关光谱学中。部分氘代类似物可用作质谱应用中的分子量分布标签，其中R1 =未取代或取代的烷基，包括环烷基或杂环烷基; R 2 = H，未取代或取代的烷基，包括环烷基或杂环烷基，或未取代或取代的芳基或杂芳基，或这些基团的任何组合; X = CH2，C = O，C = NORa，C = NNRaNRb，CH（ORa），O，S，SO，SO2或这些基团的任何其他衍生物，其中Ra和Rb独立地为H或有机残基，特别是未取代或取代的（环）烷基或杂环烷基，未取代或取代的芳基或杂芳基; Z =带负电荷的基团，每个阴离子具有1,2,3,4或5个电荷。

3. 发明申请

US20070047287A1 Method and apparatus for storing a three-dimensional arrangement of data bits in a solid-state body 审中-公开
标题翻译：用于在固体中存储数据位的三维排列的方法和装置
公开(公告)号：US20070047287A1
公开(公告)日：2007-03-01
申请号：US11510194
申请日：2006-08-25
申请人： Stefan Hell , Stefan Jakobs , Martin Andresen , Andre-C Stiel , Christian Eggeling
发明人： Stefan Hell , Stefan Jakobs , Martin Andresen , Andre-C Stiel , Christian Eggeling
IPC分类号： G11C11/00
CPC分类号： G11B7/0045 , B82Y10/00 , G11B7/244 , G11B7/246 , G11B2007/0009 , G11B2007/24624
摘要： A method which serves for writing a three-dimensional arrangement of data bits to a solid-state body comprises the steps of selecting a protein having fluorescence properties that can be altered by means of an optical write signal; providing the solid-state body made from the protein, the protein being present in the solid-state body in crystalline form; setting a spatial distribution which corresponds to the three-dimensional arrangement of data bits of the fluorescence properties of the protein of the solid-state body by means of the optical write signal.
摘要翻译：用于将数据位的三维排列写入固态体的方法包括以下步骤：选择具有可通过光写入信号改变的荧光特性的蛋白质; 提供由蛋白质制成的固态体，该蛋白质以结晶形式存在于固态体中; 通过光写入信号设置与固体物质的蛋白质的荧光特性的数据位的三维排列相对应的空间分布。

4. 发明申请

US20110160083A1 HIGH SPATIAL RESOLUTION IMAGING OF A STRUCTURE OF INTEREST IN A SPECIMEN 有权
标题翻译：一个样本中利益结构的高空间分辨率成像
公开(公告)号：US20110160083A1
公开(公告)日：2011-06-30
申请号：US13013359
申请日：2011-01-25
申请人： Stefan Hell , Christian Eggeling , Alexander Egner , Jonas Fölling , Andreas Schönle , Mariano Bossi
发明人： Stefan Hell , Christian Eggeling , Alexander Egner , Jonas Fölling , Andreas Schönle , Mariano Bossi
IPC分类号： C40B30/04 , C40B30/10
CPC分类号： G01N21/6458 , G01N21/6428 , G02B21/16 , G02B21/367 , G02B27/58
摘要： For imaging of a structure, the structure is marked with a substance which can be converted by a switching signal from a first into a second state, and which provides an optical measurement signal in one of its states, only. The switching signal is applied such that at least 10% of the molecules of the substance being in the measurement signal providing state are at a distance from their closest neighbors, which is greater than the spatial resolution limit of imaging the specimen onto a sensor array, which in turn is greater than an average distance between the molecules of the substance. From an intensity distribution of the measurement signal recorded with the sensor array, the position is only determined for those molecules of the substance which are at a distance from their closest neighboring molecules in the measurement signal providing state, which is greater than the spatial resolution limit.
摘要翻译：对于结构的成像，结构用可以通过开关信号从第一状态转换为第二状态并且仅在其状态之一提供光学测量信号的物质标记。施加开关信号，使得处于测量信号提供状态的物质的至少10％的分子距离它们最近的邻近物一个距离，其大于将样本成像到传感器阵列上的空间分辨率极限，其又大于物质分子之间的平均距离。根据记录有传感器阵列的测量信号的强度分布，该位置仅对于在测量信号提供状态中离它们最近的相邻分子一定距离的物质的那些分子确定，其大于空间分辨率极限。

5. 发明授权

US07719679B2 Method and device for optically measuring a sample 失效
标题翻译：用于光学测量样品的方法和装置
公开(公告)号：US07719679B2
公开(公告)日：2010-05-18
申请号：US11955793
申请日：2007-12-13
申请人： Stefan Hell , Christian Eggeling , Gerald Donnert
发明人： Stefan Hell , Christian Eggeling , Gerald Donnert
IPC分类号： G01N21/64
CPC分类号： G01N21/6458
摘要： The inventive method for optically measuring a sample consists in temporarily repeatedly transmitting an electromagnetic signal (2) to the sample in such a way that a substance contained in the sample is transferred from a first electronic state (1) into a second electronic state (3), wherein at least one part of said substance in the second state (3) emits photons which are used for carrying out the optical measurement of the sample, the signal (2) is transmitted to the same sample area at a certain repetition interval and said repetition interval of the signal (2) is adjusted with a lifetime of the second state (3) of the substance having an order of magnitude of 1 ns on a value of at least 0.1 μs which is optimized with respect to photon yield from the substance.
摘要翻译：用于光学测量样品的本发明的方法在于将样品中包含的物质从第一电子状态（1）转移到第二电子状态（3）的方式临时重复地将电磁信号（2）发送到样品），其中在第二状态（3）中的所述物质的至少一部分发射用于进行样品的光学测量的光子，所述信号（2）以一定的重复间隔传输到相同的样品区域，所述信号（2）的所述重复间隔在具有1ns量级的物质的第二状态（3）的寿命的至少0.1μs的值上被调整，所述寿命针对来自物质。

6. 发明授权

US07880150B2 High spatial resolution imaging of a structure of interest in a specimen 有权
标题翻译：样本中感兴趣结构的高空间分辨率成像
公开(公告)号：US07880150B2
公开(公告)日：2011-02-01
申请号：US12128948
申请日：2008-05-29
申请人： Stefan Hell , Christian Eggeling , Alexander Egner , Jonas Fölling , Andreas Schönle , Mariano Bossi
发明人： Stefan Hell , Christian Eggeling , Alexander Egner , Jonas Fölling , Andreas Schönle , Mariano Bossi
IPC分类号： G01N21/64
CPC分类号： G01N21/6458 , G01N21/6428 , G02B21/16 , G02B21/367 , G02B27/58
摘要： For the high spatial resolution imaging of a structure of interest in a specimen, a substance is selected from a group of substances which have a fluorescent first state and a nonfluorescent second state; which can be converted fractionally from their first state into their second state by light which excites them into fluorescence, and which return from their second state into their first state; the specimen's structure of interest is imaged onto a sensor array, a spatial resolution limit of the imaging being greater (i.e. worse) than an average spacing between closest neighboring molecules of the substance in the specimen; the specimen is exposed to light in a region which has dimensions larger than the spatial resolution limit, fractions of the substance alternately being excited by the light to emit fluorescent light and converted into their second state, and at least 10% of the molecules of the substance that are respectively in the first state lying at a distance from their closest neighboring molecules in the first state which is greater than the spatial resolution limit; and the fluorescent light, which is spontaneously emitted by the substance from the region, is registered in a plurality of images recorded by the sensor array during continued exposure of the specimen to the light.
摘要翻译：对于样本中感兴趣的结构的高空间分辨率成像，物质选自具有荧光第一状态和非荧光第二状态的物质组; 其可以通过将它们激发成荧光的光从其第一状态分数转换成第二状态，并且从它们的第二状态返回到其第一状态; 将感兴趣的标本的结构成像到传感器阵列上，成像的空间分辨率极限比样品中物质的最接近的相邻分子之间的平均间距更大（即更差）试样在具有大于空间分辨率极限的尺寸的区域中暴露于光，该物质的部分交替地被光激发以发射荧光并转换成其第二状态，并且至少10％的分别处于第一状态的物质，其处于与第一状态相比最大相邻分子一定距离，该距离大于空间分辨率极限; 并且由该区域内的物质自发地发出的荧光被记录在传感器阵列记录的多个图像中，该样本在样品持续暴露于光下。

7. 发明申请

US20080088839A1 Method and Device for Optically Measuring a Sample 失效
标题翻译：用于光学测量样品的方法和装置
公开(公告)号：US20080088839A1
公开(公告)日：2008-04-17
申请号：US11955793
申请日：2007-12-13
申请人： Stefan Hell , Christian Eggeling , Gerald Donnert
发明人： Stefan Hell , Christian Eggeling , Gerald Donnert
IPC分类号： G01J3/30 , G01N21/64
CPC分类号： G01N21/6458
摘要： The inventive method for optically measuring a sample consists in temporarily repeatedly transmitting an electromagnetic signal (2) to the sample in such a way that a substance contained in the sample is transferred from a first electronic state (1) into a second electronic state (3), wherein at least one part of said substance in the second state (3) emits photons which are used for carrying out the optical measurement of the sample, the signal (2) is transmitted to the same sample area at a certain repetition interval and said repetition interval of the signal (2) is adjusted with a lifetime of the second state (3) of the substance having an order of magnitude of 1 ns on a value of at least 0.1 μs which is optimized with respect to photon yield from the substance.
摘要翻译：用于光学测量样品的本发明的方法在于将样品中包含的物质从第一电子状态（1）转移到第二电子状态（3）的方式临时重复地将电磁信号（2）发送到样品），其中在第二状态（3）中的所述物质的至少一部分发射用于进行样品的光学测量的光子，所述信号（2）以一定的重复间隔传输到相同的样品区域，所述信号（2）的所述重复间隔在具有1ns量级的物质的第二状态（3）的寿命的情况下调整为至少0.1μA的值，其相对于来自物质。

8. 发明授权

US08084754B2 High spatial resolution imaging of a structure of interest in a specimen 有权
标题翻译：样本中感兴趣结构的高空间分辨率成像
公开(公告)号：US08084754B2
公开(公告)日：2011-12-27
申请号：US13013359
申请日：2011-01-25
申请人： Stefan Hell , Christian Eggeling , Alexander Egner , Jonas Fölling , Andreas Schönle , Mariano Bossi
发明人： Stefan Hell , Christian Eggeling , Alexander Egner , Jonas Fölling , Andreas Schönle , Mariano Bossi
IPC分类号： G01N21/64
CPC分类号： G01N21/6458 , G01N21/6428 , G02B21/16 , G02B21/367 , G02B27/58
摘要： For imaging of a structure, the structure is marked with a substance which can be converted by a switching signal from a first into a second state, and which provides an optical measurement signal in one of its states, only. The switching signal is applied such that at least 10% of the molecules of the substance being in the measurement signal providing state are at a distance from their closest neighbors, which is greater than the spatial resolution limit of imaging the specimen onto a sensor array, which in turn is greater than an average distance between the molecules of the substance. From an intensity distribution of the measurement signal recorded with the sensor array, the position is only determined for those molecules of the substance which are at a distance from their closest neighboring molecules in the measurement signal providing state, which is greater than the spatial resolution limit.
摘要翻译：对于结构的成像，结构用可以通过开关信号从第一状态转换为第二状态并且仅在其状态之一提供光学测量信号的物质标记。施加开关信号，使得处于测量信号提供状态的物质的至少10％的分子距离它们最近的邻近物一个距离，其大于将样本成像到传感器阵列上的空间分辨率极限，其又大于物质分子之间的平均距离。根据记录有传感器阵列的测量信号的强度分布，该位置仅对于在测量信号提供状态中离它们最近的相邻分子一定距离的物质的那些分子确定，其大于空间分辨率极限。

9. 发明申请

US20090134342A1 High spatial resolution imaging of a structure of interest in a specimen 有权
标题翻译：样本中感兴趣结构的高空间分辨率成像
公开(公告)号：US20090134342A1
公开(公告)日：2009-05-28
申请号：US12128948
申请日：2008-05-29
申请人： Stefan Hell , Christian Eggeling , Alexander Egner , Jonas Folling , Andreas Schonle , Mariano Bossi
发明人： Stefan Hell , Christian Eggeling , Alexander Egner , Jonas Folling , Andreas Schonle , Mariano Bossi
IPC分类号： G01J1/58 , G01N1/30
CPC分类号： G01N21/6458 , G01N21/6428 , G02B21/16 , G02B21/367 , G02B27/58
摘要： For the high spatial resolution imaging of a structure of interest in a specimen, a substance is selected from a group of substances which have a fluorescent first state and a nonfluorescent second state; which can be converted fractionally from their first state into their second state by light which excites them into fluorescence, and which return from their second state into their first state; the specimen's structure of interest is imaged onto a sensor array, a spatial resolution limit of the imaging being greater (i.e. worse) than an average spacing between closest neighboring molecules of the substance in the specimen; the specimen is exposed to light in a region which has dimensions larger than the spatial resolution limit, fractions of the substance alternately being excited by the light to emit fluorescent light and converted into their second state, and at least 10% of the molecules of the substance that are respectively in the first state lying at a distance from their closest neighboring molecules in the first state which is greater than the spatial resolution limit; and the fluorescent light, which is spontaneously emitted by the substance from the region, is registered in a plurality of images recorded by the sensor array during continued exposure of the specimen to the light.
摘要翻译：对于样本中感兴趣的结构的高空间分辨率成像，物质选自具有荧光第一状态和非荧光第二状态的物质组; 其可以通过将它们激发成荧光的光从其第一状态分数转换成第二状态，并且从它们的第二状态返回到其第一状态; 将感兴趣的标本的结构成像到传感器阵列上，成像的空间分辨率极限比样品中物质的最接近的相邻分子之间的平均间距更大（即更差）试样在具有大于空间分辨率极限的尺寸的区域中暴露于光，该物质的部分交替地被光激发以发射荧光并转换成其第二状态，并且至少10％的分别处于第一状态的物质，其处于与第一状态相比最大相邻分子一定距离，该距离大于空间分辨率极限; 并且由该区域内的物质自发地发出的荧光被记录在传感器阵列记录的多个图像中，该样本在样品持续暴露于光下。

10. 发明申请

US20080018891A1 METHOD OF PRODUCING SPATIAL FINE STRUCTURES 有权
标题翻译：生产空间细微结构的方法
公开(公告)号：US20080018891A1
公开(公告)日：2008-01-24
申请号：US11856887
申请日：2007-09-18
申请人： Stefan Hell , Volker Westphal , Norbert Quaas
发明人： Stefan Hell , Volker Westphal , Norbert Quaas
IPC分类号： G01J3/30
CPC分类号： G01N21/6458 , G01N21/6489 , G01N21/91 , G01N21/956 , G02B21/0076 , G03F7/105 , G03F7/70616 , G03F7/70675
摘要： A method of producing spatial fine structures comprises the steps of: selecting a luminophore from the group of luminophores displaying two different states, one of the two states being an active state in which luminescence light is obtainable from the luminophore, the other of the two states being an inactive state in which no luminescence light is obtainable from the luminophore, and the luminophore being reversibly, but essentially completely, transferable out the one state into the other state by means of an optical signal; adding the luminophore to a material; forming a spatial fine structure of the material; and fluorescence-microscopically examining whether the desired fine structure is present. The step of fluorescence-microscopically examining comprises the sub-steps of: outside measuring points of interest, transferring the luminophore into the other state with the optical signal, the luminophore being essentially completely transferred into the inactive state outside the measuring points, and measuring luminescence light only emitted by the luminophore in the active state, to even resolve lines of the fine structure at a distance of less than 100 nm.
摘要翻译：一种产生空间精细结构的方法包括以下步骤：从显示两种不同状态的发光体组中选择发光体，两种状态中的一种状态是可从发光体获得发光的活性状态，两种状态中的另一种状态是其中从发光体不能获得发光的无活性状态，并且发光体是可逆的，但基本上完全可通过光信号转移到一个状态到另一状态; 将发光体添加到材料中; 形成材料的空间精细结构; 并进行荧光显微镜检查是否存在所需的精细结构。荧光显微镜检查的步骤包括以下子步骤：在感兴趣的测量点之外，用光信号将发光体转移到另一状态，发光体基本上完全转移到测量点外的无效状态，并测量发光发光体仅在活性状态下发光，甚至在小于100nm的距离处解决精细结构的线。

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