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1. 发明申请

US20140147836A1 METHODS AND APPARATUS FOR IMAGING MOLECULES IN LIVING SYSTEMS 审中-公开
标题翻译：用于在生活系统中成像分子的方法和装置
公开(公告)号：US20140147836A1
公开(公告)日：2014-05-29
申请号：US13820528
申请日：2011-09-13
申请人： David Grunwald , Robert H. Singer
发明人： David Grunwald , Robert H. Singer
IPC分类号： G01N21/64
CPC分类号： G01N21/6486 , G01N21/6458 , G01N2021/6421 , G01N2021/6441
摘要： Methods and apparatus are disclosed for imaging molecular interactions in living cells at high resolution, low light levels and high acquisition speeds.
摘要翻译：公开了用于以高分辨率，低光级和高采集速度对活细胞中的分子相互作用进行成像的方法和装置。

2. 发明申请

US20110318732A1 PREDICTION OF CHEMOTHERAPEUTIC RESPONSE VIA SINGLE-CELL PROFILING OF TRANSCRIPTION SITE ACTIVATION 审中-公开
标题翻译：通过单细胞表达转录酶活性预测化疗反应
公开(公告)号：US20110318732A1
公开(公告)日：2011-12-29
申请号：US12736885
申请日：2009-05-07
申请人： Robert H. Singer , Rosanna Pezo , Leonard H. Augenlicht
发明人： Robert H. Singer , Rosanna Pezo , Leonard H. Augenlicht
IPC分类号： C12Q1/68
CPC分类号： G01N33/5011 , C12Q1/6886 , C12Q2600/106 , C12Q2600/158
摘要： The present invention generally relates to methods for determining tumor resistance or sensitivity to chemotherapeutic agents and the likelihood of tumor reoccurrence based on the expression levels of genes known to correlate to the chemotherapeutic agent. In particular, the expression levels of TYMS, MRGX, ATP7B and/or BAK in tumor cells, as measured by the number of active transcription sites detected by fluorescence in situ hybridization (FISH), are predictive of resistance and sensitivity to chemotherapy and the likelihood of reoccurrence following chemotherapy treatment.
摘要翻译：本发明一般涉及用于确定肿瘤抗性或对化学治疗剂的敏感性的方法以及基于已知与化学治疗剂相关的基因的表达水平的肿瘤再发生的可能性。特别地，通过荧光原位杂交（FISH）检测到的活性转录位点的数量，肿瘤细胞中的TYMS，MRGX，ATP7B和/或BAK的表达水平预示着对化疗的抗性和敏感性以及可能性化疗后复发。

3. 发明授权

US06534266B1 Assay of transcription sites by multi-fluor fish 失效
标题翻译：通过多氟鱼测定转录位点
公开(公告)号：US06534266B1
公开(公告)日：2003-03-18
申请号：US09556134
申请日：2000-04-21
申请人： Robert H. Singer
发明人： Robert H. Singer
IPC分类号： C12Q168
CPC分类号： C12Q1/6841 , C12Q2565/102 , C12Q2563/185 , C12Q2563/107 , C12Q2537/143
摘要： An in situ hybridization method for detecting and specifically identifying transcription of a multiplicity of different target sequences in a cell is disclosed. The method includes assigning a different bar code to at least five target sequences, with each target sequence containing at least one predetermined subsequence. Each bar code contains at least one fluorochrome, and at least one bar code comprises at least two different, spectrally distinguishable fluorochromes. A probe set specific for each target sequence is provided in the method. Each probe set contains a hybridization probe complementary to each subsequence in the target sequence. Each probe is labeled with a fluorochrome, and the fluorochromes in each probe set collectively correspond to the bar code for the target sequence of that probe set.
摘要翻译：公开了用于检测和特异性鉴定细胞中多种不同靶序列的转录的原位杂交方法。该方法包括将不同的条形码分配给至少五个目标序列，每个目标序列包含至少一个预定的子序列。每个条形码包含至少一个荧光染料，并且至少一个条形码包含至少两个不同的光谱可区分的荧光染料。在该方法中提供了针对每个靶序列特异的探针组。每个探针组包含与目标序列中每个亚序列互补的杂交探针。每个探针用荧光染料标记，并且每个探针组中的荧光染料集合对应于该探针组的靶序列的条形码。

4. 发明授权

US5827660A Caged fluorochrome-labeled probes and subtraction of autoflourescence background 失效
标题翻译：笼状荧光染料标记的探针和自动鉴定背景的减法
公开(公告)号：US5827660A
公开(公告)日：1998-10-27
申请号：US693786
申请日：1996-08-09
申请人： Robert H. Singer , Joan C. Politz
发明人： Robert H. Singer , Joan C. Politz
IPC分类号： C07H19/10 , C12Q1/68 , G01N15/00 , C12Q1/70 , G01N1/00 , G01N33/53
CPC分类号： C07H19/10 , C12Q1/6816 , C12Q1/6841 , G01N2015/0065 , G01N2021/6439
摘要： Disclosed are methods for removing autofluorescence background from microscopic images or from flow cytometer measurements. The microscopic images can be fluorescent in situ hybridization images or images of living cells. The methods involve the steps of: (1) detecting fluorescence from a cell before "uncaging" a caged fluorochrome label on a probe; (2) uncaging the fluorochrome; (3) detecting fluorescence from the cell; and (4) subtracting fluorescence detected before uncaging from fluorescence detected after uncaging. Also disclosed is a method for tracking the movement of a target molecule in a living cell. Also disclosed is a fluorochrome-uncaging flow cytometer. The fluorochrome-uncaging flow cytometer includes a first fluorescence excitation light beam, an uncaging light beam, a second fluorescence excitation light beam, an electronic data system, a first photodetector operably linked to the electronic data system, and a second photodetector operably linked to the electronic data system.
摘要翻译：公开了从微观图像或流式细胞仪测量中去除自体荧光背景的方法。微观图像可以是荧光原位杂交图像或活细胞图像。所述方法包括以下步骤：（1）在探针上“笼养”笼状荧光染料标记之前检测来自细胞的荧光; （2）褪色荧光染料; （3）检测来自细胞的荧光; 和（4）在解开之后从荧光检测到的荧光减去脱色之前检测到的荧光。还公开了用于跟踪活细胞中靶分子运动的方法。还公开了荧光染料 - 血液流式细胞仪。荧光染料流式细胞仪包括第一荧光激发光束，解吸光束，第二荧光激发光束，电子数据系统，可操作地连接到电子数据系统的第一光电检测器，以及可操作地连接到电子数据系统的第二光电检测器电子数据系统。

5. 发明授权

US06203986B1 Visualization of RNA in living cells 有权
标题翻译：活细胞中RNA的可视化
公开(公告)号：US06203986B1
公开(公告)日：2001-03-20
申请号：US09177268
申请日：1998-10-22
申请人： Robert H. Singer , Edouard Bertrand
发明人： Robert H. Singer , Edouard Bertrand
IPC分类号： A61B1000
CPC分类号： G01N33/5035 , C07K14/005 , C07K14/43595 , C07K2319/00 , C07K2319/09 , C07K2319/42 , C07K2319/735 , C12N15/62 , C12N2795/18122 , C12Q1/6841 , C12Q1/6844 , G01N33/5008 , G01N33/5308
摘要： A method for visualizing the location and movement of a specific RNA of interest in a living cell, in real time, is disclosed. The method includes the following steps: (a) providing a DNA encoding the RNA, which RNA includes a protein-binding site; (b) providing a nucleic acid encoding a fusion protein, which fusion protein comprises a fluorescent domain and an RNA-binding domain; (c) introducing the DNA encoding the RNA, and the nucleic acid encoding the fusion protein, into a eukaryotic cell so that the DNA encoding the RNA and the nucleic acid encoding the fusion protein are expressed in the cell; and (d) detecting fluorescence outside the nucleus or inside the nucleus of the cell, with the fluorescence being from the fusion protein bound to the RNA. In some embodiments, the fusion protein also includes an intracellular localization domain.
摘要翻译：公开了一种用于实时观察活细胞中感兴趣的特定RNA的位置和运动的方法。该方法包括以下步骤：（a）提供编码RNA的DNA，该RNA包括蛋白质结合位点; （b）提供编码融合蛋白的核酸，所述融合蛋白包含荧光结构域和RNA结合结构域; （c）将编码RNA的DNA和编码融合蛋白的核酸引入真核细胞，使得编码RNA的DNA和编码融合蛋白的核酸在细胞中表达; 和（d）检测细胞核外的荧光或细胞核内的荧光，荧光来自与RNA结合的融合蛋白。在一些实施方案中，融合蛋白还包括细胞内定位结构域。

6. 发明授权

US5866331A Single molecule detection by in situ hybridization 失效
标题翻译：通过原位杂交检测单分子
公开(公告)号：US5866331A
公开(公告)日：1999-02-02
申请号：US546072
申请日：1995-10-20
申请人： Robert H. Singer , Andrea M. Femino
发明人： Robert H. Singer , Andrea M. Femino
IPC分类号： C12Q1/68 , G01J1/58 , G02B21/34
CPC分类号： C12Q1/6841 , C12Q1/6816
摘要： Disclosed are methods for accurately determining the total emission intensity of a single fluorochrome, under imaging conditions, using a digital imaging fluorescence microscopy system. Also disclosed are methods for detecting and localizing probe-target molecule binding. The detection method has sufficient resolution and sensitivity to locate and detect a single target-bound probe.
摘要翻译：公开了使用数字成像荧光显微镜系统在成像条件下精确测定单个荧光染料的总发光强度的方法。还公开了用于检测和定位探针 - 靶分子结合的方法。检测方法具有足够的分辨率和灵敏度来定位和检测单个靶结合探针。

7. 发明授权

US5641675A Cis-acting sequences for intracellular localization of RNA 失效
标题翻译：用于RNA的细胞内定位的顺式作用序列
公开(公告)号：US5641675A
公开(公告)日：1997-06-24
申请号：US319836
申请日：1994-10-07
申请人： Robert H. Singer , Edward H. Kislauskis
发明人： Robert H. Singer , Edward H. Kislauskis
IPC分类号： C07K14/47 , C12N15/85 , C12N5/00 , C07H21/04
CPC分类号： C07K14/4716 , C12N15/85
摘要： Disclosed are the particular 3' noncoding nucleotide sequences (zipcode and zipcode elements) that cause .beta.-actin mRNA (and .beta.-actin protein synthesis) to be localized in specific regions of the cytoplasm. Also disclosed is the discovery that zipcode function can be fully inhibited by antisense oligonucleotides.
摘要翻译：公开了引起β-actin mRNA（和β-肌动蛋白蛋白合成）定位于细胞质特定区域的特定3'非编码核苷酸序列（zipcode和zipcode元件）。还公开了这样的发现：反义寡核苷酸可以完全抑制zipcode功能。

8. 发明授权

US5523204A Detection of nucleic acids in cells by strand displacement amplification 失效
标题翻译：通过链置换扩增检测细胞中的核酸
公开(公告)号：US5523204A
公开(公告)日：1996-06-04
申请号：US165719
申请日：1993-12-10
申请人： Robert H. Singer , Jean Marie Mathys , Kenton L. Lohman
发明人： Robert H. Singer , Jean Marie Mathys , Kenton L. Lohman
IPC分类号： C12N15/09 , C12N15/00 , C12Q1/68 , C12Q1/70 , C12R1/92 , C12N15/34
CPC分类号： C12Q1/6844 , C12Q1/6841 , C12Q1/6881 , C12Q1/703
摘要： Strand Displacement Amplification for amplification of nucleic acid target sequences in situ in cells in suspension, on slides or in tissues. Excellent specimen morphology is preserved, and either DNA targets, or RNA targets, or both may be selectively amplified. In situ amplification by SDA is compatible with immunochemical techniques, so that both amplification of target sequences and immunological staining can be performed on the same specimen.
摘要翻译：链置换扩增用于在悬浮液，载玻片或组织中的细胞中原位扩增核酸靶序列。保留了良好的标本形态，并且可以选择性地扩增DNA靶或RNA靶，或两者。 SDA的原位扩增与免疫化学技术相容，可以在同一标本上进行靶序列的扩增和免疫染色。

9. 发明授权

US06586240B1 Visualization of RNA in living cells 失效
标题翻译：活细胞中RNA的可视化
公开(公告)号：US06586240B1
公开(公告)日：2003-07-01
申请号：US09656010
申请日：2000-09-06
申请人： Robert H. Singer , Edouard Bertrand
发明人： Robert H. Singer , Edouard Bertrand
IPC分类号： C12N1585
CPC分类号： G01N33/5035 , C07K14/005 , C07K14/43595 , C07K2319/00 , C07K2319/09 , C07K2319/42 , C07K2319/735 , C12N15/62 , C12N2795/18122 , C12Q1/6841 , C12Q1/6844 , G01N33/5008 , G01N33/5308
摘要： A method for visualizing the location and movement of a specific RNA of interest in a living cell, in real time, is disclosed. The method includes the following steps: (a) providing a DNA encoding the RNA, which RNA includes a protein-binding site; (b) providing a nucleic acid encoding a fusion protein, which fusion protein comprises a fluorescent domain and an RNA-binding domain; (c) introducing the DNA encoding the RNA, and the nucleic acid encoding the fusion protein, into a eukaryotic cell so that the DNA encoding the RNA and the nucleic acid encoding the fusion protein are expressed in the cell; and (d) detecting fluorescence outside the nucleus or inside the nucleus of the cell, with the fluorescence being from the fusion protein bound to the RNA. In some embodiments, the fusion protein also includes an intracellular localization domain.
摘要翻译：公开了一种用于实时观察活细胞中感兴趣的特定RNA的位置和运动的方法。该方法包括以下步骤：（a）提供编码RNA的DNA，该RNA包括蛋白质结合位点; （b）提供编码融合蛋白的核酸，所述融合蛋白包含荧光结构域和RNA结合结构域; （c）将编码RNA的DNA和编码融合蛋白的核酸引入真核细胞，使得编码RNA的DNA和编码融合蛋白的核酸在细胞中表达; 和（d）检测细胞核外的荧光或细胞核内的荧光，荧光来自与RNA结合的融合蛋白。在一些实施方案中，融合蛋白还包括细胞内定位结构域。

10. 发明授权

US5962332A Detection of trinucleotide repeats by in situ hybridization 失效
标题翻译：通过原位杂交检测三核苷酸重复序列
公开(公告)号：US5962332A
公开(公告)日：1999-10-05
申请号：US570155
申请日：1995-12-11
申请人： Robert H. Singer , Krishan L. Taneja
发明人： Robert H. Singer , Krishan L. Taneja
IPC分类号： C12Q1/68 , G01N33/00 , C07H21/04 , G01N21/76
CPC分类号： C12Q1/6883 , C12Q1/6818 , C12Q1/6827 , C12Q1/6841 , C12Q2600/156 , C12Q2600/158 , Y10S436/80 , Y10S436/805 , Y10T436/143333
摘要： Disclosed is a method of detecting a trinucleotide repeat expansion by in situ hybridization. The disclosed method uses a sample of nucleated cells, a labeled trinucleotide repeat-specific probe and detection of the hybridized probe by a means whose sensitivity distinguishes between the signal from probes hybridized to an expanded repeat and the signal from probes hybridized to a non-expanded repeat.
摘要翻译：公开了通过原位杂交检测三核苷酸重复扩增的方法。所公开的方法使用有核细胞的样品，标记的三核苷酸重复特异性探针和通过灵敏度区分来自与扩增重复杂交的探针的信号的灵敏度区别的杂交探针，以及来自与未扩增的杂交的探针的信号重复。

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