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71. 发明授权

US5625048A Modified green fluorescent proteins 失效
标题翻译：改良的绿色荧光蛋白
公开(公告)号：US5625048A
公开(公告)日：1997-04-29
申请号：US337915
申请日：1994-11-10
申请人： Roger Y. Tsien , Roger Heim
发明人： Roger Y. Tsien , Roger Heim
IPC分类号： C12N15/09 , C07K14/435 , C12N5/10 , C12N15/12 , C12N15/65 , C12P21/04 , C12Q1/25 , C12Q1/68 , G01N33/533 , C07H21/04
CPC分类号： C07K14/43595 , C12N15/65 , G01N33/533 , G01N2333/43595 , Y10S530/855
摘要： Modifications in the sequence of Aequorea wild-type GFP provide products having markedly different excitation and emission spectra from corresponding products from wild-type GFP. In one class of modifications, the product derived from the modified GFP exhibits an alteration in the ratio of two main excitation peaks observed with the product derived from wild-type GFP. In another class, the product derived from the modified GFP fluoresces at a shorter wavelength than the corresponding product from wild-type GFP. In yet another class of modifications, the product derived from the modified GFP exhibits only a single excitation peak and enhanced emission relative to the product derived from wild-type GFP.
摘要翻译： Aequorea野生型GFP序列的修饰提供了来自野生型GFP相应产物的产生明显不同的激发和发射光谱的产物。在一类修饰中，衍生自经修饰的GFP的产物表现出与用野生型GFP产生的产物观察到的两个主要激发峰的比例的变化。在另一类中，衍生自经修饰的GFP的产物在比来自野生型GFP的相应产物更短的波长处发荧光。在另一类修饰中，衍生自经修饰的GFP的产物相对于衍生自野生型GFP的产物仅显示单个激发峰和增强的发射。

72. 发明授权

US5602258A Azide-containing chelators whose affinity for calcium ion is decreased by illumination 失效
标题翻译：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。

73. 发明授权

US5283433A Scanning confocal microscope providing a continuous display 失效
标题翻译：扫描共焦显微镜提供连续显示
公开(公告)号：US5283433A
公开(公告)日：1994-02-01
申请号：US958038
申请日：1992-10-05
申请人： Roger Y. Tsien
发明人： Roger Y. Tsien
IPC分类号： G01N21/64 , G02B21/00 , H01J3/14
CPC分类号： G02B21/008 , G02B21/0076
摘要： A scanning confocal microscope scans a sample with an incident beam of radiation, in a raster scan pattern, causing the sample to fluoresce and emit visible radiation in at least two wavelengths. A portion of the fluorescent light retraces a portion of the path optical of incident beam, to a dichroic mirror that separates it from the incident beam for detection by a pair of photomultiplier tubes. A data processor accumulates digital data from the photomultiplier tubes to form a succession of image data frames of the sample being scanned, in the two wavelengths. Image data for a selected number of frames in each wavelength is averaged and then recorded on a single track of a recording system, in an alternating fashion with averaged data for the other wavelength. In addition, a ratio of the averaged data for the two wavelengths is delivered to a video display.
摘要翻译：扫描共焦显微镜以光栅扫描图案中的入射光束扫描样品，导致样品在至少两个波长内发出可见光。荧光的一部分将入射光束的光路的一部分返回到分色镜，该分色镜将其与入射光束分离，以便由一对光电倍增管检测。数据处理器累积来自光电倍增管的数字数据，以在两个波长内形成扫描样本的一系列图像数据帧。对每个波长中所选择的帧数的图像数据进行平均化，然后以与其他波长的平均数据交替的方式记录在记录系统的单个轨道上。此外，两个波长的平均数据的比率被传送到视频显示器。

74. 发明授权

US5134232A Fluorescent indicator dyes for alkali metal cations 失效
标题翻译：用于碱金属阳离子的荧光指示剂染料
公开(公告)号：US5134232A
公开(公告)日：1992-07-28
申请号：US391879
申请日：1989-08-09
申请人： Roger Y. Tsien , Akwasi Minta
发明人： Roger Y. Tsien , Akwasi Minta
IPC分类号： C07D273/00 , C07D273/08 , C07D413/04 , C07D413/14 , C07D491/04 , G01N33/84
CPC分类号： C07D273/00 , C07D413/04 , C07D413/14 , C07D491/04 , G01N33/84
摘要： The invention discloses fluorescent chelator compounds that are especially useful to monitor and measure cytosolic concentrations of alkali metal cations such as Na.sup.+, K.sup.+, and Li.sup.+. The new compounds are comprised of: (1) crown ethers (that may or may not have substituent groups attached to the core carbons, but will always contain at least one core nitrogen) that are linked via the core nitrogen(s) to at least one (2) fluorophore that contains an additional heteroaromatic liganding center. In the currently preferred dye, SBFI, the core compound is crown ether 1,7-diaza-4,10,13-trioxacyclopentadecane and the heteroaromatic fluorophores are benzofurans that are linked to isophthalate groups. Selectivities for Na.sup.+ over K.sup.+ of about 20 are observed, resulting in effective dissociation constants for Na.sup.+ of about 20 mM against a background of 120 mM K.sup.+. Increasing [Na.sup.+ ] increases the ratio of excitation efficiency at 330-345 nm to that at 370-390 nm with emission collected at 450-550 nm, so that ratio fluorometry and imaging work at the same wavelengths as used with the well-known Ca.sup.2+ indicator fura-2. If the macrocyclic ring is increased in size to a 1,10-diaza-4,7,13,16-tetraoxacyclooctadecane, the chelators become selective for K.sup.+ over Na.sup.+. If the ring is decreased in size, for example to a 1,7-diaza-4,10-dioxacyclododecane, the chelators become selective for Li.sup.+ over Na.sup.+.
摘要翻译：本发明公开了特别可用于监测和测量碱金属阳离子如Na +，K +和Li +的胞质浓度的荧光螯合剂化合物。新化合物包括：（1）冠醚（其可以具有或不具有连接到核心碳上的取代基但总是含有至少一个核心氮），其通过核心氮至少连接至少一个（2）荧光团，其中包含一个额外的杂芳族结合中心。在目前优选的染料中，SBFI，核心化合物是冠醚1,7二氮杂-4,10,13-三氧杂环十五烷，杂芳族荧光团是与间苯二酸酯基团连接的苯并呋喃。观察到Na +在K +上约20的选择性，导致对于120mM K +背景的Na +约20mM的有效解离常数。增加[Na +]增加了在330-345nm处的激发效率与在370-390nm的激发效率的比率，其中在450-550nm收集的发射，使得比例荧光测定和成像工作在与公知的Ca2 + 指标fura-2。如果大环的大小增加到1,10-二氮杂-4,7,13,16-四氧杂环十八烷，则螯合剂对于Na +而言对K +变得选择性。如果环尺寸减小，例如减小到1,7-diaza-4,10-二氧杂环十二烷，那么螯合剂对Na +的选择性变得更高。

75. 发明授权

US4689432A Chelators whose affinity for calcium is decreased by illumination 失效
标题翻译：通过照明降低钙对钙的亲和力的螯合剂
公开(公告)号：US4689432A
公开(公告)日：1987-08-25
申请号：US648536
申请日：1984-09-07
申请人： Roger Y. Tsien , Grzegorz Grynkiewicz , Akwasi Minta
发明人： Roger Y. Tsien , Grzegorz Grynkiewicz , Akwasi Minta
IPC分类号： C07F9/09 , C07C67/00 , C07C201/00 , C07C205/37 , C07C227/00 , C07C227/16 , C07C229/16 , C07C229/18 , C07C301/00 , C07C309/66 , C07D215/40 , C07D317/62 , C07D319/18 , C07F9/60 , C09K3/00 , G01N33/84 , C07C101/26 , C07D215/38
CPC分类号： G01N33/84 , C07D317/62 , Y10S424/06
摘要： The present invention discloses a group of calcium chelating compounds which have a descreased affinity for calcium following illumination. These new compounds contain a photolabile nitrobenzyl derivative coupled to a tetracarboxylate Ca.sup.2+ chelating parent compound having the octacoordinate chelating groups characteristic of EGTA or BAPTA. In a first form, the new compounds are comprised of a BAPTA-like chelator coupled to a single 2-nitrobenzyl derivative, which in turn is a photochemical precursor of a 2-nitrosobenzophenone. In a second form, the new compounds are comprised of a BAPTA-like chelator coupled to two 2-nitrobenzyl derivatives, themselves photochemical prcursors of the related 2-nitrosobenzophenones.The present invention also discloses a novel method for preparing 1-hydroxy- or 1-alkoxy-1-(2-nitroaryl)-1-aryl methanes. Methanes of this type are critical to the preparation of, or actually constitute, the photolabile Ca.sup.2+ chelating compounds disclosed and claimed herein.
摘要翻译：本发明公开了一种钙照射后对钙的亲和力降低的钙螯合化合物。这些新化合物含有偶联到具有EGTA或BAPTA特征的八配位螯合基团的四羧酸盐Ca 2+螯合母体化合物的光不稳定的硝基苄基衍生物。在第一种形式中，新化合物由与单一2-硝基苄基衍生物偶联的BAPTA样螯合剂组成，其又是2-亚硝基二苯甲酮的光化学前体。在第二种形式中，新化合物由与两个2-硝基苄基衍生物偶联的BAPTA样螯合剂组成，它们本身是相关的2-亚硝基二苯甲酮的光化学原子。本发明还公开了一种制备1-羟基 - 或1-烷氧基-1-（2-硝基芳基）-1-芳基甲烷的新方法。这种类型的甲烷对于制备或实际构成本文公开和要求保护的光不稳定的Ca 2+螯合化合物至关重要。

76. 发明授权

US08685372B2 Peptides and aptamers for targeting of neuron or nerves 有权
标题翻译：用于靶向神经元或神经的肽和适体
公开(公告)号：US08685372B2
公开(公告)日：2014-04-01
申请号：US13264512
申请日：2010-04-15
申请人： Roger Y. Tsien , Quyen T. Nguyen , Michael Whitney
发明人： Roger Y. Tsien , Quyen T. Nguyen , Michael Whitney
IPC分类号： A61K49/00 , G01N33/53 , A61P25/00 , C07K19/00 , C07K17/02 , A61K38/18 , C07K7/08 , C07K17/10
CPC分类号： C12N15/115 , A61K38/00 , A61K49/0032 , A61K49/0054 , C07K7/08 , C12N2310/16 , G01N33/582
摘要： The present invention provides methods for guiding preservation of neurons or nerves during surgery by administering a fluorescently-labeled peptide or aptamer that specifically binds to the neurons or nerves. The invention further provides targeting molecules of fluorescently-labeled peptides or aptamers that specifically bind to neurons or nerves and for compositions thereof.
摘要翻译：本发明提供了通过施用特异性结合神经元或神经的荧光标记的肽或适体来指导手术期间神经元或神经保护的方法。本发明还提供了特异性结合神经元或神经的荧光标记肽或适体的靶向分子及其组合物。

77. 发明授权

US08679742B2 Method for determining the presence of an enzyme in a sample 失效
标题翻译：测定样品中酶的存在的方法
公开(公告)号：US08679742B2
公开(公告)日：2014-03-25
申请号：US12612580
申请日：2009-11-04
申请人： Roger Y. Tsien , Roger Heim , Andrew Cubitt
发明人： Roger Y. Tsien , Roger Heim , Andrew Cubitt
IPC分类号： C07K14/00
CPC分类号： C12Q1/485 , C07K14/43595 , C07K2319/00 , C12Q1/37 , G01N33/542 , G01N2333/43595
摘要： This invention provides tandem fluorescent protein construct including a donor fluorescent protein moiety, an acceptor fluorescent protein moiety and a linker moiety that couples the donor and acceptor moieties. The donor and acceptor moieties exhibit fluorescence resonance energy transfer which is eliminated upon cleavage. The constructs are useful in enzymatic assays.
摘要翻译：本发明提供了包括供体荧光蛋白部分，受体荧光蛋白部分和连接供体和受体部分的接头部分的串联荧光蛋白构建体。供体和受体部分表现出在切割时消除的荧光共振能量转移。所述构建体可用于酶测定。

78. 发明申请

US20130330718A1 PROTEINS THAT EFFICIENTLY GENERATE SINGLET OXYGEN 有权
标题翻译：蛋白质能有效地产生单一氧气
公开(公告)号：US20130330718A1
公开(公告)日：2013-12-12
申请号：US13992540
申请日：2011-12-07
申请人： Xiaokun Shu , Roger Y. Tsien
发明人： Xiaokun Shu , Roger Y. Tsien
IPC分类号： G01N21/64 , C07K14/00
CPC分类号： G01N21/6486 , C07K14/001 , C07K2319/00 , C12N9/12 , C12Y207/11001
摘要： The present invention provides miniSOG proteins, polynucleotides, and methods of use. When expressed in a bacterial or mammalian cell, miniSOG proteins spontaneously incorporate flavin mononucleotide and produce fluorescence and singlet oxygen upon excitation. Uses include optical and electron microscope imaging, in vivo imaging, detection and localization of protein-protein interactions, and photoablation.
摘要翻译：本发明提供了miniSOG蛋白，多核苷酸和使用方法。当在细菌或哺乳动物细胞中表达时，miniSOG蛋白自发并入黄素单核苷酸，并在激发时产生荧光和单线态氧。用途包括光学和电子显微镜成像，体内成像，蛋白质 - 蛋白质相互作用的检测和定位以及光消融。

79. 发明申请

US20120148499A1 PEPTIDES AND APTAMERS FOR TARGETING OF NEURON OR NERVES 有权
标题翻译：用于治疗神经或神经的肽和蛋白酶
公开(公告)号：US20120148499A1
公开(公告)日：2012-06-14
申请号：US13264512
申请日：2010-04-15
申请人： Roger Y. Tsien , Quyen T. Nguyen , Michael Whitney
发明人： Roger Y. Tsien , Quyen T. Nguyen , Michael Whitney
IPC分类号： A61K49/00 , G01N33/53 , A61K38/10 , A61P25/00 , C07K19/00 , C07K17/02 , A61K38/18 , C07K7/08 , C07K17/10
CPC分类号： C12N15/115 , A61K38/00 , A61K49/0032 , A61K49/0054 , C07K7/08 , C12N2310/16 , G01N33/582
摘要： The present invention provides methods for guiding preservation of neurons or nerves during surgery by administering a fluorescently-labeled peptide or aptamer that specifically binds to the neurons or nerves. The invention further provides targeting molecules of fluorescently-labeled peptides or aptamers that specifically bind to neurons or nerves and for compositions thereof.
摘要翻译：本发明提供了通过施用特异性结合神经元或神经的荧光标记的肽或适体来指导手术期间神经元或神经保护的方法。本发明还提供了特异性结合神经元或神经的荧光标记肽或适体的靶向分子及其组合物。

80. 发明授权

US08071761B2 Substrates for beta-lactamase and uses thereof 有权
标题翻译： β-内酰胺酶底物及其用途
公开(公告)号：US08071761B2
公开(公告)日：2011-12-06
申请号：US11606642
申请日：2006-11-29
申请人： Roger Y. Tsien , Gregor Zlokarnik
发明人： Roger Y. Tsien , Gregor Zlokarnik
IPC分类号： C07D501/14
CPC分类号： C07D501/36 , C07D501/00 , C07D505/00 , C07D519/00 , C12N9/86 , C12N15/85 , C12N2800/90 , C12N2830/002 , C12P17/184 , C12P17/188 , C12P35/00 , C12Q1/18 , C12Q1/34 , C12Q1/6897 , C12Q2334/00 , G01N2333/986 , G01N2415/00 , Y10S435/805
摘要： Substrates for β-lactamase of the general formula I in which one of X and Y is a fluorescent donor moiety and the other is a quencher (which may or may not re-emit); R′ is selected from the group consisting of H, lower (i.e., alkyl of 1 to about 5 carbon atoms) and (CH2)nOH, in which n is 0 or an integer from 1 to 5; R″ is selected from the group consisting of H, physiologically acceptable metal and ammonium cations, —CHR2OCO(CH2)nCH3, —CHR2OCOC (CH3)3, acylthiomethyl, acyloxy-alpha-benzyl, delta-butyrolactonyl, methoxycarbonyloxymethyl, phenyl, methylsulphinylmethyl, beta-morpholinoethyl, dialkylaminoethyl, acyloxyalkyl, dialkylaminocarbonyloxymethyl and aliphatic, in which R2 is selected from the group consisting of H and lower alkyl; A is selected from the group consisting of S, O, SO, SO2 and CH2; and Z′ and Z″ are linkers for the fluorescent donor and quencher moieties. Methods of assaying β-lactamase activity and monitoring expression in systems using β-lactamase as a reporter gene also are disclosed.
摘要翻译： X和Y中的一个是荧光供体部分，另一个是猝灭剂（其可以或不再发射）的通式I的β-内酰胺酶底物; R'选自H，低级（即1至约5个碳原子的烷基）和（CH 2）nOH，其中n为0或1至5的整数; -CH 2 COO（CH 2）n CH 3，-CHR 2 OCOC（CH 3）3，酰基硫代甲基，酰氧基-α-苄基，δ-丁基内酰胺，甲氧基羰氧基甲基，苯基，甲基亚磺酰基甲基， β-吗啉基乙基，二烷基氨基乙基，酰氧基烷基，二烷基氨基羰氧基甲基和脂族基，其中R2选自H和低级烷基; A选自S，O，SO，SO 2和CH 2; 和Z'和Z“是荧光供体和猝灭剂部分的接头。公开了使用β-内酰胺酶作为报道基因的系统中测定β-内酰胺酶活性和监测表达的方法。

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