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    High spatial resolution imaging of a structure of interest in a specimen
    3.
    发明授权
    High spatial resolution imaging of a structure of interest in a specimen 有权
    样本中感兴趣结构的高空间分辨率成像

    公开(公告)号：US08174692B2

    公开(公告)日：2012-05-08

    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： G01J3/30

    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。

    NOVEL HYDROPHILIC AND LIPOPHILIC RHODAMINES FOR LABELLING AND IMAGING
    4.
    发明申请
    NOVEL HYDROPHILIC AND LIPOPHILIC RHODAMINES FOR LABELLING AND IMAGING 有权
    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。

    Hydrophilic and lipophilic rhodamines for labelling and imaging
    5.
    发明授权
    Hydrophilic and lipophilic rhodamines for labelling and imaging 有权
    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。

    HIGH SPATIAL RESOLUTION IMAGING OF A STRUCTURE OF INTEREST IN A SPECIMEN
    6.
    发明申请
    HIGH SPATIAL RESOLUTION IMAGING OF A STRUCTURE OF INTEREST IN A SPECIMEN 有权
    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 。

    High spatial resolution imaging of a structure of interest in a specimen
    7.
    发明授权
    High spatial resolution imaging of a structure of interest in a specimen 有权
    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 。

    High spatial resolution imaging of a structure of interest in a specimen
    8.
    发明申请
    High spatial resolution imaging of a structure of interest in a specimen 有权
    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。

    High spatial resolution imaging of a structure of interest in a specimen
    9.
    发明授权
    High spatial resolution imaging of a structure of interest in a specimen 有权
    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。