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    Detection of analytes using fluorescent energy transfer
    1.
    发明授权
    Detection of analytes using fluorescent energy transfer 失效
    使用荧光能量转移检测分析物

    公开(公告)号：US5439797A

    公开(公告)日：1995-08-08

    申请号：US114103

    申请日：1993-08-30

    申请人： Roger Y. Tsien ,  Susan S. Taylor ,  Stephen R. Adams ,  Ying Ji

    发明人： Roger Y. Tsien ,  Susan S. Taylor ,  Stephen R. Adams ,  Ying Ji

    IPC分类号： C12N9/00 ,  C12Q1/48 ,  G01N21/64 ,  G01N33/542 ,  G01N33/58 ,  G01N33/68 ,  G01N33/533

    CPC分类号： G01N33/542 ,  G01N33/582 ,  Y10S436/80

    摘要： The invention provides labeled proteins suitable for determining the presence of cAMP, other second messengers and organic molecules. The proteins are separately labeled with fluorochromes which, when in close spatial proximity, preferably, less than about 6 nm, interact through the transfer of energy from one fluorochrome to the other.A composition of matter, (S.sub.1.A).sub.n1 (S.sub.2.D).sub.n2 is provided wherein S.sub.1 and S.sub.2 are two proteins which are associated in one state and substantially disassociated in another, the equilibrium between which is controlled by the free concentration of an analyte, and A and D are fluorochromes, the emission wavelength of fluorochrome D overlapping the excitation wavelength of fluorochrome A and the distance between A and D being in sufficiently close proximity to allow the radiationless transfer of energy between the fluorochromes. A and D can be selected from a variety of acceptor, donor pairs such as fluorescein and tetramethylrhodamine and derivatives thereof. The concentration of analytes such as cAMP in a sample can be determined by contacting the sample with (S.sub.1.A).sub.n1 (S.sub.2.D).sub.n2 providing energy near the excitation wavelength of D and measuring the fluorescence of A or D, the concentration of cAMP and other said analytes being determined by the ratio of emission of D to the emission of A, previously calibrated with reference solutions of known analyte concentration.

    摘要翻译： 本发明提供了适合于测定cAMP，其他第二信使和有机分子的存在的标记蛋白质。 蛋白质分别用荧光染料标记，当空间接近时，优选小于约6nm，通过将能量从一个荧光染料转移到另一个荧光染料相互作用。 提供了一种物质组合物（S1.A）n1（S2.D）n2，其中S1和S2是两种蛋白质，两种蛋白质在一种状态下相互关联，并且在另一种状态下基本上脱离，其中的平衡受自由浓度 分析物，A和D是荧光染料，荧光染料D的发射波长与荧光染料A的激发波长重叠，A和D之间的距离足够接近，从而允许在荧光染料之间无辐射传递能量。 A和D可以选自多种受体，诸如荧光素和四甲基罗丹明的供体对及其衍生物。 通过使样品与（S1.A）n1（S2.D）n2接触，在D的激发波长附近提供能量并测量A或D的荧光，可以测定样品中cAMP等分析物的浓度， cAMP和其他所述分析物由D的排放与A的排放比例决定，其先前用已知分析物浓度的参考溶液校准。

    Chelators whose affinity for calcium ion is increased by illumination
    2.
    发明授权
    Chelators whose affinity for calcium ion is increased by illumination 失效
    通过照明增加对钙离子的亲和力的螯合剂

    公开(公告)号：US5141627A

    公开(公告)日：1992-08-25

    申请号：US412852

    申请日：1989-09-25

    申请人： Roger Y. Tsien ,  Stephen R. Adams

    发明人： Roger Y. Tsien ,  Stephen R. Adams

    IPC分类号： C07C245/18

    CPC分类号： C07C245/18

    摘要： The present invention relates to a group of organic chelators whose affinity for calcium ion in solution is increased by electromagnetic radiation. Specifically, the chelators are related to BAPTA and utilize the addition of an electron-withdrawing group (e.g., diazocarbonyl) to a ring of BAPTA, para to the amino group. Photochemical rearrangement of the diazoacetyl group converts the group to the electron-donating carboxymethyl group, causing the calcium ion efficiency to increase 25 to 50 fold. These chelators when incorporated into rat fibroblasts either by microinjection or by incubation as the membrane-permeable, enzymatically-labile tetraacetoxymethyl ester and flash-photolyzed cause a drop in intracellular free calcine ion to or below resting valves of about 10.sup.-7 M. These chelators are used to generate controlled fast removal of intracellular free calcium ion to mimic or modulate a number of important cellular responses, especially in nerve or muscle.

    摘要翻译： 本发明涉及通过电磁辐射增加溶液中对钙离子的亲和力的一组有机螯合剂。 具体地说，螯合剂与BAPTA有关，并利用向吸附对氨基的BAPTA环中加入吸电子基团（如重氮羰基）。 重氮乙酰基的光化学重排将该基团转移到给电子的羧甲基基团，导致钙离子效率增加25-50倍。 这些螯合剂通过显微注射或通过孵育作为膜可渗透的，酶促不稳定的四乙酰氧基甲基酯并且闪光光引发而引入大鼠成纤维细胞时，导致细胞内游离的煅烧离子降至约10-7M的静止阀或其下面。这些螯合剂 用于产生受控的快速去除细胞内游离钙离子以模拟或调节许多重要的细胞反应，特别是在神经或肌肉中。

    Azide-containing chelators whose affinity for calcium ion is decreased by illumination
    3.
    发明授权
    Azide-containing chelators whose affinity for calcium ion is decreased by illumination 失效
    含钙螯合剂，其对钙离子的亲和力通过照射而降低

    公开(公告)号：US5552555A

    公开(公告)日：1996-09-03

    申请号：US475047

    申请日：1995-06-06

    申请人： Roger Y. Tsien ,  Stephen R. Adams

    发明人： Roger Y. Tsien ,  Stephen R. Adams

    IPC分类号： C07D277/28 ,  C07D413/04 ,  C07D413/14

    CPC分类号： C07D413/04 ,  C07D413/14

    摘要： The present invention relates to a group of organic chelators whose affinity for calcium ion in solution is decreased by electromagnetic radiation. Specifically, the chelators are related to fura-2 and utilize the addition of an azide group to the 3-position of the benzofuran ring of a fura-2 type structure. Photolysis of the azide group causes the calcium ion affinity to decrease 100 to 1000 fold. These chelators when incorporated into rat fibroblasts either by microinjection or by incubation as the membrane-permeable, enzymatically-labile esters and flash-photolyzed cause large increases in intracellular free calcium ion. These chelators are used to generate controlled fast elevation of intracellular free calcium ion concentration to mimic or modulate a number of important cellular responses, especially in nerve or muscle.

    摘要翻译： 本发明涉及通过电磁辐射降低溶液中对钙离子的亲和力的一组有机螯合剂。 具体地说，螯合剂与fura-2相关，并且利用在fura-2型结构的苯并呋喃环的3-位上添加叠氮基。 叠氮化物基团的光解使钙离子亲和力降低100〜1000倍。 当这些螯合剂通过显微注射或通过温育作为膜可渗透的酶不稳定的酯并且闪光光解引入大鼠成纤维细胞时，引起细胞内游离钙离子的大量增加。 这些螯合剂用于产生受控的快速升高的细胞内游离钙离子浓度以模拟或调节许多重要的细胞反应，特别是在神经或肌肉中。

    Light-triggered indicators that memorize analyte concentrations
    4.
    发明授权
    Light-triggered indicators that memorize analyte concentrations 失效
    记录分析物浓度的光触发指标

    公开(公告)号：US06180411B2

    公开(公告)日：2001-01-30

    申请号：US09134668

    申请日：1998-07-30

    申请人： Roger Y. Tsien ,  Stephen R. Adams

    发明人： Roger Y. Tsien ,  Stephen R. Adams

    IPC分类号： G01N33532

    CPC分类号： C07D307/82 ,  G01N33/84 ,  Y10S436/805

    摘要： A new class of optical indicators which are capable of memorizing and preserving the spatial localization of intracellular analytes in a time resolved manner is described. The compounds comprise a chromophore carrying a photolabile group capable of undergoing an irreversible and detectable chemical transformation upon irradiation by light. The chromophore is linked to a binding site capable of binding an analyte, wherein binding of the analyte to the binding site alters an optical property of the chromophore, thus altering the ability of the photolabile group to undergo the chemical transformation. Methods and kits for memorizing the spatial localization of the analytes are also described.

    摘要翻译： 描述了能够以时间分辨方式记忆和保留细胞内分析物的空间定位的新类型的光学指示器。 所述化合物包含携带光照不稳定基团的发色团，所述光不稳定基团能在光照射时经历不可逆和可检测的化学转化。 发色团连接到能够结合分析物的结合位点，其中分析物与结合位点的结合改变发色团的光学性质，从而改变光不稳定组进行化学转化的能力。 还描述了用于记忆分析物的空间定位的方法和试剂盒。

    Azide-containing chelators whose affinity for calcium ion is decreased by illumination
    5.
    发明授权
    Azide-containing chelators whose affinity for calcium ion is decreased by illumination 失效
    含钙螯合剂，其对钙离子的亲和力通过照射而降低

    公开(公告)号：US5602258A

    公开(公告)日：1997-02-11

    申请号：US968002

    申请日：1992-10-23

    申请人： Roger Y. Tsien ,  Stephen R. Adams

    发明人： Roger Y. Tsien ,  Stephen R. Adams

    IPC分类号： C07D277/28 ,  C07D413/04 ,  C07D413/14 ,  C07D263/30

    CPC分类号： C07D413/04 ,  C07D413/14

    摘要： The present invention relates to a group of organic chelators whose affinity for calcium ion in solution is decreased by electromagnetic radiation. Specifically, the chelators are related to fura-2 and utilize the addition of an azide group to the 3-position of the benzofuron ring of a fura-2 type structure. Photolysis of the azide group causes the calcium ion affinity to decrease 100 to 1000 fold. These chelators when incorporated into rat fibroblasts either by microinjection or by incubation as the membrane-permeable, enzymatically-labile esters and flash-photolyzed cause large increases in intracellular free calcium ion. These chelators are used to generate controlled fast elevation of intracellular free calcium ion concentration to mimic or modulate a number of important cellular responses, especially in nerve or muscle.

    摘要翻译： 本发明涉及通过电磁辐射降低溶液中对钙离子的亲和力的一组有机螯合剂。 具体地说，螯合剂与fura-2相关，并且利用向fura-2型结构的苯并呋喃环的3位添加叠氮基。 叠氮化物基团的光解使钙离子亲和力降低100〜1000倍。 当这些螯合剂通过显微注射或通过温育作为膜可渗透的酶不稳定的酯并且闪光光解引入大鼠成纤维细胞时，引起细胞内游离钙离子的大量增加。 这些螯合剂用于产生受控的快速升高的细胞内游离钙离子浓度以模拟或调节许多重要的细胞反应，特别是在神经或肌肉中。

    System and method for performing path-sensitive value flow analysis on a program
    6.
    发明授权
    System and method for performing path-sensitive value flow analysis on a program 有权
    对程序执行路径敏感值流分析的系统和方法

    公开(公告)号：US07089537B2

    公开(公告)日：2006-08-08

    申请号：US10662942

    申请日：2003-09-15

    申请人： Manuvir Das ,  Stephen R. Adams ,  Nurit Dor

    发明人： Manuvir Das ,  Stephen R. Adams ,  Nurit Dor

    IPC分类号： G06F9/44

    CPC分类号： G06F8/433

    摘要： Described is a method and system for performing path-sensitive value flow analysis on a software program. Concrete state and value alias information is tracked along each statement and each relevant path in an abstract program and is stored as a symbolic state in a symbolic store. The value alias information includes a first set of aliases that identify aliases for a designated value that is being analyzed and a second set of aliases that identify possible aliases for the designated value. The value alias information is obtained using imprecise memory alias analysis. Along each relevant path for each statement, transforms are applied to the sets of aliases to update the first and second sets of aliases. The transforms are applied based on the type of statement being processed. Symbolic states existing at the same location are merged if the value alias information is identical within the symbolic states.

    摘要翻译： 描述了一种用于在软件程序上执行路径敏感值流分析的方法和系统。 每个语句和抽象程序中的每个相关路径跟踪具体状态和值别名信息，并将其作为符号状态存储在符号存储中。 值别名信息包括识别被分析的指定值的别名的第一组别名以及标识指定值的可能别名的第二组别名。 使用不精确的内存别名分析获取值别名信息。 沿着每个语句的每个相关路径，转换将应用于别名集以更新第一和第二组别名。 转换根据正在处理的语句的类型应用。 如果符号状态中的值别名信息相同，则在同一位置存在的符号状态将被合并。