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公开(公告)号：US09448231B2
公开(公告)日：2016-09-20
申请号：US13402567
申请日：2012-02-22
申请人： Chol Steven Yun , Brian Daniel Kelly , Julia Ashworth-Sharpe , Christopher A. Bieniarz , Pascal Bamford , Adrian E. Murillo
发明人： Chol Steven Yun , Brian Daniel Kelly , Julia Ashworth-Sharpe , Christopher A. Bieniarz , Pascal Bamford , Adrian E. Murillo
CPC分类号： G01N33/5308 , B82Y15/00 , C07D487/04 , C09B57/08 , C09B69/10 , C09B69/109 , C12Q1/6816 , C12Q1/6886 , C12Q2600/158 , G01N33/588 , C12Q2563/107 , C12Q2563/155
摘要： Embodiments of a system, method, and kit for visualizing a nucleus are disclosed. A tissue sample is pretreated with a protease to permeabilize the nucleus, and then incubated with a nanoparticle/DNA-binding moiety conjugate. The DNA-binding moiety includes at least one DNA-binding molecule. The conjugate binds to DNA within the nucleus, and the nanoparticle is visualized, thereby visualizing the nucleus. Computer and image analysis techniques are used to evaluate nuclear features such as chromosomal distribution, ploidy, shape, size, texture features, and/or contextual features. The method may be used in combination with other multiplexed tests on the tissue sample, including fluorescence in situ hybridization. Kits for performing the method include a protease enzyme composition, a nanoparticle/DNA-binding moiety conjugate, and a reaction buffer.
摘要翻译： 公开了用于可视化核的系统，方法和试剂盒的实施例。 用蛋白酶预处理组织样品以使细胞核透化，然后与纳米颗粒/ DNA结合部分缀合物一起温育。 DNA结合部分包括至少一个DNA结合分子。 该结合物与细胞核内的DNA结合，并且纳米颗粒被可视化，从而可视化细胞核。 计算机和图像分析技术用于评估核特征，如染色体分布，倍性，形状，大小，纹理特征和/或语境特征。 该方法可以与组织样品上的其他复用测试结合使用，包括荧光原位杂交。 用于进行该方法的试剂盒包括蛋白酶组合物，纳米颗粒/ DNA结合部分缀合物和反应缓冲液。
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2.发明授权Substrates for chromogenic detection and methods of use in detection assays and kits 有权用于显色检测的底物和用于检测测定和试剂盒的方法公开(公告)号：US09127028B2
公开(公告)日：2015-09-08
申请号：US13809354
申请日：2011-08-12
IPC分类号： C07F9/12 , C12Q1/70 , C07F9/6503 , G01N21/78
CPC分类号： C07F9/65031 , G01N21/78
摘要： Embodiments of substrates and processes for chromogenic detection, and in particular pyrazolyl dihydrogen phosphate compounds, are disclosed.
摘要翻译： 公开了用于显色检测的底物和方法，特别是吡唑基二氢磷酸酯化合物的实施方案。
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3.发明授权Method for chromogenic detection of two or more target molecules in a single sample 有权一种样品中两种或更多种靶分子的显色检测方法公开(公告)号：US08481270B2
公开(公告)日：2013-07-09
申请号：US13059274
申请日：2009-08-21
申请人： Richard Gniewek , Michael Farrell , Hiroaki Nitta , Megan Lehrkamp , Jerome Kosmeder , Brian Daniel Kelly , Thomas Grogan , Fabien Gaire , Mary Padilla , Christopher Bieniarz
发明人： Richard Gniewek , Michael Farrell , Hiroaki Nitta , Megan Lehrkamp , Jerome Kosmeder , Brian Daniel Kelly , Thomas Grogan , Fabien Gaire , Mary Padilla , Christopher Bieniarz
CPC分类号： C12Q1/6841 , G01N33/53 , G01N33/581 , G01N33/583
摘要： The present invention provides a method and kit for detection of two or more target molecules in a single tissue sample, such as for gene and protein dual detection in a single tissue sample. Methods comprise treating a tissue sample with a first binding moiety that specifically binds a first target molecule. Methods further comprise treating the tissue sample with a solution containing a soluble electron-rich aromatic compound prior to or concomitantly with contacting the tissue sample with a hapten-labeled binding moiety and detecting a second target molecule. In one example, the first target molecule is a protein and the second is a nucleic acid sequence, the first target molecule being detected by immunohistochemistry and the second by in situ hybridization. The disclosed method reduces background due to non-specific binding of the hapten-labeled specific binding moiety to an insoluble electron rich compound deposited near the first target molecule.
摘要翻译： 本发明提供了用于检测单个组织样品中两种或更多种靶分子的方法和试剂盒，例如用于单一组织样品中的基因和蛋白质双重检测。 方法包括用特异性结合第一靶分子的第一结合部分处理组织样品。 方法还包括在使组织样品与半抗原标记的结合部分接触或同时检测第二靶分子之前或同时使用含有可溶于富电子的芳族化合物的溶液处理组织样品。 在一个实例中，第一靶分子是蛋白质，第二个是核酸序列，第一个靶分子通过免疫组织化学检测，第二个是原位杂交。 所公开的方法由于半抗原标记的特异性结合部分与沉积在第一靶分子附近的不溶性富电子化合物的非特异性结合而减少背景。
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公开(公告)号：US20120219948A1
公开(公告)日：2012-08-30
申请号：US13402567
申请日：2012-02-22
申请人： Chol Steven Yun , Brian Daniel Kelly , Julia Ashworth-Sharpe , Christopher A. Bieniarz , Pascal Bamford , Adrian E. Murillo
发明人： Chol Steven Yun , Brian Daniel Kelly , Julia Ashworth-Sharpe , Christopher A. Bieniarz , Pascal Bamford , Adrian E. Murillo
IPC分类号： C12Q1/68 , G01N21/64 , C07D493/04 , C07D471/04 , B82Y5/00 , B82Y15/00
CPC分类号： G01N33/5308 , B82Y15/00 , C07D487/04 , C09B57/08 , C09B69/10 , C09B69/109 , C12Q1/6816 , C12Q1/6886 , C12Q2600/158 , G01N33/588 , C12Q2563/107 , C12Q2563/155
摘要： Embodiments of a system, method, and kit for visualizing a nucleus are disclosed. A tissue sample is pretreated with a protease to permeabilize the nucleus, and then incubated with a nanoparticle/DNA-binding moiety conjugate. The DNA-binding moiety includes at least one DNA-binding molecule. The conjugate binds to DNA within the nucleus, and the nanoparticle is visualized, thereby visualizing the nucleus. Computer and image analysis techniques are used to evaluate nuclear features such as chromosomal distribution, ploidy, shape, size, texture features, and/or contextual features. The method may be used in combination with other multiplexed tests on the tissue sample, including fluorescence in situ hybridization. Kits for performing the method include a protease enzyme composition, a nanoparticle/DNA-binding moiety conjugate, and a reaction buffer.
摘要翻译： 公开了用于可视化核的系统，方法和试剂盒的实施例。 用蛋白酶预处理组织样品以使细胞核透化，然后与纳米颗粒/ DNA结合部分缀合物一起温育。 DNA结合部分包括至少一个DNA结合分子。 该结合物与细胞核内的DNA结合，并且纳米颗粒被可视化，从而可视化细胞核。 计算机和图像分析技术用于评估核特征，如染色体分布，倍性，形状，大小，纹理特征和/或语境特征。 该方法可以与组织样品上的其他复用测试结合使用，包括荧光原位杂交。 用于进行该方法的试剂盒包括蛋白酶组合物，纳米颗粒/ DNA结合部分缀合物和反应缓冲液。
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5.发明申请SUBSTRATES FOR CHROMOGENIC DETECTION AND METHODS OF USE IN DETECTION ASSAYS AND KITS 有权用于染色体检测的基质和检测方法和试剂盒公开(公告)号：US20130115593A1
公开(公告)日：2013-05-09
申请号：US13809354
申请日：2011-08-12
CPC分类号： C07F9/65031 , G01N21/78
摘要： Embodiments of substrates and processes for chromogenic detection, and in particular pyrazolyl dihydrogen phosphate compounds, are disclosed.
摘要翻译： 公开了用于显色检测的底物和方法，特别是吡唑基二氢磷酸酯化合物的实施方案。
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6.发明申请METHOD FOR CHROMOGENIC DETECTION OF TWO OR MORE TARGET MOLECULES IN A SINGLE SAMPLE 有权用于在单个样品中检测两种或更多种目标分子的方法公开(公告)号：US20110136130A1
公开(公告)日：2011-06-09
申请号：US13059274
申请日：2009-08-21
申请人： Richard Gniewek , Michael Farrell , Hiroaki Nitta , Megan Lehrkamp , Jerome Kosmeder , Brian Daniel Kelly , Fabien Gaire , Mary Padilla , Thomas Grogan , Christopher Bieniarz
发明人： Richard Gniewek , Michael Farrell , Hiroaki Nitta , Megan Lehrkamp , Jerome Kosmeder , Brian Daniel Kelly , Fabien Gaire , Mary Padilla , Thomas Grogan , Christopher Bieniarz
CPC分类号： C12Q1/6841 , G01N33/53 , G01N33/581 , G01N33/583
摘要： The present invention provides a method and kit for detection of two or more target molecules in a single tissue sample, such as for gene and protein dual detection in a single tissue sample. Methods comprise treating a tissue sample with a first binding moiety that specifically binds a first target molecule. Methods further comprise treating the tissue sample with a solution containing a soluble electron-rich aromatic compound prior to or concomitantly with contacting the tissue sample with a hapten-labeled binding moiety and detecting a second target molecule. In one example, the first target molecule is a protein and the second is a nucleic acid sequence, the first target molecule being detected by immunohistochemistry and the second by in situ hybridization. The disclosed method reduces background due to non-specific binding of the hapten-labeled specific binding moiety to an insoluble electron rich compound deposited near the first target molecule.
摘要翻译： 本发明提供了用于检测单个组织样品中两种或更多种靶分子的方法和试剂盒，例如用于单一组织样品中的基因和蛋白质双重检测。 方法包括用特异性结合第一靶分子的第一结合部分处理组织样品。 方法还包括在使组织样品与半抗原标记的结合部分接触或同时检测第二靶分子之前或同时使用含有可溶于富电子的芳族化合物的溶液处理组织样品。 在一个实例中，第一靶分子是蛋白质，第二个是核酸序列，第一个靶分子通过免疫组织化学检测，第二个是原位杂交。 所公开的方法由于半抗原标记的特异性结合部分与沉积在第一靶分子附近的不溶性富电子化合物的非特异性结合而减少背景。
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