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1. 发明授权

US07989220B2 Metal-enhanced fluorescence for polarization-based affinity assays 有权
标题翻译：金属增强荧光用于基于偏振的亲和力测定
公开(公告)号：US07989220B2
公开(公告)日：2011-08-02
申请号：US12427722
申请日：2009-04-21
申请人： Joseph R. Lakowicz , Henryk Szmacinski
发明人： Joseph R. Lakowicz , Henryk Szmacinski
IPC分类号： G01N33/553
CPC分类号： G01N33/587 , G01N21/6428 , G01N21/6445 , G01N21/6452
摘要： A method and kit for determining the quantity of an analyte include providing a functionalized substrate and a reagent. The functionalized substrate includes metallic nanoparticles and a plurality of substantively identical bioactive target molecules affixed to a substrate. The bioactive target molecule binds to a particular analyte. The reagent includes identical detection molecules. Each detection molecule includes a fluorophore, and binds to a particular analyte or competes with a particular analyte for binding to the target molecule. The functionalized substrate is contacted to a test sample and the reagent. The functionalized substrate and a covering solution are exposed to polarized electromagnetic waves that excite the fluorophore. A quantity of the particular analyte in the test sample is determined based on measuring polarization anisotropy of fluorescent emissions from the substrate and the covering solution.
摘要翻译：用于确定分析物的量的方法和试剂盒包括提供官能化的底物和试剂。功能化底物包括金属纳米颗粒和固定到基底上的多个基本相同的生物活性靶分子。生物活性靶分子与特定分析物结合。该试剂包括相同的检测分子。每个检测分子包括荧光团，并结合特定分析物或与特定分析物竞争结合靶分子。将功能化底物与测试样品和试剂接触。将功能化基材和覆盖溶液暴露于激发荧光团的极化电磁波。基于测量来自基底和覆盖溶液的荧光发射的偏振各向异性来测定测试样品中特定分析物的量。

2. 发明授权

US06472221B1 Lifetime-based sensing of sodium and potassium 失效
标题翻译：基于终生的钠和钾感测
公开(公告)号：US06472221B1
公开(公告)日：2002-10-29
申请号：US09666721
申请日：2000-09-20
申请人： Joseph R. Lakowicz , Henryk Szmacinski
发明人： Joseph R. Lakowicz , Henryk Szmacinski
IPC分类号： C12Q100
CPC分类号： G01N21/6408 , G01N33/84 , Y10T436/25375
摘要： A system and method of optically measuring Na+ and K+ in a sample such as blood which contains high concentrations Na+ (up to 160 mM) and K+ (up to 6.5 mM) using a photoluminescent probe having intrinsic analyte-induced lifetime changes. Specifically, the use of lifetime-based sensing of Na+ and K+ at the extracellular concentrations present in whole blood or, blood serum. The preferred embodiment uses phase-modulation fluorometry.
摘要翻译：使用具有固有分析物引起的寿命变化的光致发光探针，在含有高浓度Na +（至多160mM）和K +（至多6.5mM））的样品（例如血液）中光学测量Na +和K +的系统和方法。具体来说，在全血或血清中存在的细胞外浓度使用基于寿命的Na +和K +感测。优选实施例使用相位调制荧光测定法。

3. 发明授权

US5660991A Long lifetime anisotropy (polarization) probes for clinical chemistry, immunoassays, affinity assays and biomedical research 失效
标题翻译：用于临床化学，免疫测定，亲和力测定和生物医学研究的长寿命各向异性（极化）探针
公开(公告)号：US5660991A
公开(公告)日：1997-08-26
申请号：US330743
申请日：1994-10-28
申请人： Joseph R. Lakowicz , Henryk Szmacinski , Ewald Terpetschnig
发明人： Joseph R. Lakowicz , Henryk Szmacinski , Ewald Terpetschnig
IPC分类号： G01N33/536 , G01N33/542 , G01N33/58 , G01N33/53 , G01N33/533
CPC分类号： G01N33/542 , G01N33/582
摘要： A method of conducting an immunoassay of a sample of interest is described, including the steps of (A) coupling a luminescent asymmetric metal-ligand complex to the sample of interest to form a coupled sample, (B) exciting the coupled sample with linearly polarized electromagnetic energy to cause the coupled sample to emit fluorescent light; and (C) measuring the polarization of the fluorescent emission as a measure of a biological characteristic of the sample of interest.
摘要翻译：描述了进行感兴趣的样品的免疫测定的方法，包括以下步骤：（A）将发光的不对称金属 - 配体复合物偶联到感兴趣的样品上以形成耦合的样品，（B）用线偏振电磁能量使耦合的样品发射荧光; 和（C）测量荧光发射的极化作为感兴趣样品的生物特征的量度。

4. 发明申请

US20100035335A1 METAL-ENHANCED FLUORESCENCE FOR THE LABEL-FREE DETECTION OF INTERACTING BIOMOLECULES 审中-公开
标题翻译：金属增强荧光检测交互生物分子的无标签
公开(公告)号：US20100035335A1
公开(公告)日：2010-02-11
申请号：US12537544
申请日：2009-08-07
申请人： Joseph R. Lakowicz , Henryk Szmacinski , Krishanu Ray , Mustafa Habib Chowdhury , Kazimierz Nowaczyk
发明人： Joseph R. Lakowicz , Henryk Szmacinski , Krishanu Ray , Mustafa Habib Chowdhury , Kazimierz Nowaczyk
IPC分类号： C12M1/34 , G01N21/64
CPC分类号： G01N21/6486 , G01N21/648
摘要： A method for enhancing fluorescence of a biomolecule includes the step of associating the biomolecule having intrinsic fluorescence with a sensing surface that contains nanostructured metal. Association of the biomolecule with the nanostructured metal enhances its intrinsic fluorescence, which is detected upon exposure to electromagnetic radiation of a suitable wavelength. The sensing surface may include capture or ligand molecule which binds to the biomolecule and sequesters it in proximity to the nanostructured metal, thereby causing its fluorescent signal to be enhanced. The method can be used in label-free bioassays for detection of interacting biomolecules, such as antibody-antigen binding.
摘要翻译：用于增强生物分子荧光的方法包括将具有固有荧光的生物分子与含有纳米结构金属的感测表面缔合的步骤。生物分子与纳米结构金属的结合增强其固有荧光，其在暴露于合适波长的电磁辐射时被检测。感测表面可以包括与生物分子结合并将其螯合到接近纳米结构金属的俘获或配体分子，从而导致其荧光信号被增强。该方法可用于无标记生物测定中，用于检测相互作用的生物分子，如抗体 - 抗原结合。

5. 发明授权

US5628310A Method and apparatus to perform trans-cutaneous analyte monitoring 失效
标题翻译：进行透皮分析物监测的方法和装置
公开(公告)号：US5628310A
公开(公告)日：1997-05-13
申请号：US444927
申请日：1995-05-19
申请人： Govind Rao , Henryk Szmacinski , Joseph R. Lakowicz
发明人： Govind Rao , Henryk Szmacinski , Joseph R. Lakowicz
IPC分类号： G01N21/64 , A61B5/00 , A61B5/145 , A61B5/1455 , A61B10/00
CPC分类号： A61B5/1459 , A61B5/0017 , A61B5/14532 , A61B5/14539
摘要： An apparatus and method to enable minimally invasive transdermal measurements of the fluorescence lifetime of an implanted element without reagent consumption and not requiring painful blood sampling. The monitoring apparatus displays the quantity of a selected substance present in the skin and stores the data in memory. The stored information can be transmitted via modem, or antenna, to a master station for diagnostic purposes or clinical evaluation. The use of this method and apparatus improves control of blood monitoring, and therefore, enhances long-term disease management with fewer complications.
摘要翻译：一种能够进行植入元件的荧光寿命的微创透皮测量而不需要试剂消耗并且不需要痛苦的血液取样的装置和方法。监视装置显示皮肤中存在的选定物质的量并将数据存储在存储器中。存储的信息可以通过调制解调器或天线传送到主站用于诊断目的或临床评估。这种方法和装置的使用改善了对血液监测的控制，因此以更少的并发症增强了长期的疾病管理。

6. 发明授权

US5624847A Method for optically measuring chemical analytes 失效
标题翻译：光学测量化学分析物的方法
公开(公告)号：US5624847A
公开(公告)日：1997-04-29
申请号：US102806
申请日：1993-08-06
申请人： Joseph R. Lakowicz , Henryk Szmacinski
发明人： Joseph R. Lakowicz , Henryk Szmacinski
IPC分类号： G01N15/14 , G01N21/64 , G01N31/22 , G01N33/49 , G01N33/52 , G01N33/58 , G01N33/84 , G01N21/80
CPC分类号： G01N33/52 , G01N21/6408 , G01N31/223 , G01N33/4925 , G01N33/582 , G01N33/84
摘要： A system and method in which a photoluminescent ligand is added to a sample to be analyzed in the form of a photoluminescent probe having intrinsic analyte-induced lifetime changes. The method preferably employs phase-modulation fluorometry to measure the lifetime changes. Specific probes are disclosed for measuring various analytes, particularly ionic solutes, including H.sup.+, Ca.sup.2+ and K.sup.+.
摘要翻译：将光致发光配体以具有固有分析物诱导的寿命变化的光致发光探针的形式加入到待分析样品中的系统和方法。该方法优选采用相位调制荧光测定来测量寿命变化。公开了用于测量各种分析物，特别是离子溶质（包括H +，Ca 2+和K +）的特异性探针。

7. 发明授权

US5485530A Method and apparatus for multi-dimensional phase fluorescence lifetime imaging 失效
标题翻译：多维相荧光寿命成像的方法和装置
公开(公告)号：US5485530A
公开(公告)日：1996-01-16
申请号：US094016
申请日：1993-07-23
申请人： Joseph R. Lakowicz , Klaus W. Berndt , Kazimierz Nowaczyk , Henryk Szmacinski , Michael L. Johnson
发明人： Joseph R. Lakowicz , Klaus W. Berndt , Kazimierz Nowaczyk , Henryk Szmacinski , Michael L. Johnson
IPC分类号： G01K11/20 , G01N15/14 , G01N21/64 , G06K9/00
CPC分类号： G01K11/20 , G01N21/6408 , G01N15/1427 , G01N2015/144 , G01N2015/1454 , G01N2021/6439 , G01N21/6456
摘要： A method and apparatus for detection and/or measurement of physical characteristics of a sample based on multi-dimensional phase-modulation fluorescence lifetime imaging using at least one fluorescent probe having known and/or variable fluorescent lifetimes.
摘要翻译： PCT No.PCT / US92 / 00242 Sec。 371日期：1993年7月23日 102（e）日期1993年7月23日PCT提交1993年7月23日PCT公布。出版物WO92 / 13265 日期：1992年8月6日。一种用于使用具有已知和/或可变荧光寿命的至少一个荧光探针，基于多维相位调制荧光寿命成像来检测和/或测量样品的物理特性的方法和装置。

8. 发明申请

US20170168048A1 PLASMONIC SUBSTRATES FOR METAL-ENHANCED FLUORESCENCE BASED SENSING, IMAGING AND ASSAYS FOR CELLS 审中-公开
公开(公告)号：US20170168048A1
公开(公告)日：2017-06-15
申请号：US14965560
申请日：2015-12-10
申请人： Henryk Szmacinski , Joseph R. Lakowicz
发明人： Henryk Szmacinski , Joseph R. Lakowicz
IPC分类号： G01N33/553 , G01N21/552 , G01N21/64 , G01N33/68 , G01N33/543
CPC分类号： G01N33/553 , G01N21/274 , G01N21/6408 , G01N21/648 , G01N33/54346
摘要： Techniques for metal enhanced fluorescence include determining a calibration curve that relates concentration of a particular analyte to at least one of intensity or lifetime of fluorescent emissions at a plasmonic substrate in response to incident light, for a plurality of known concentrations of the particular analyte mixed with a reagent. The reagent comprises a detection molecule. A concentration of the particular analyte in a vicinity of a cell in a sample is determined directly from the calibration curve and measurements, in response to the incident light, of at least one of intensity or lifetime of fluorescent emissions at the plasmonic substrate in contact with the cell and reagent.

9. 发明申请

US20090275145A1 METAL-ENHANCED FLUORESCENCE FOR POLARIZATION-BASED AFFINITY ASSAYS 有权
标题翻译：金属增强荧光的基于极化的亲和力测定
公开(公告)号：US20090275145A1
公开(公告)日：2009-11-05
申请号：US12427722
申请日：2009-04-21
申请人： Joseph R. Lakowicz , Henryk Szmacinski
发明人： Joseph R. Lakowicz , Henryk Szmacinski
IPC分类号： G01N33/566 , G01N21/76 , G01N33/553
CPC分类号： G01N33/587 , G01N21/6428 , G01N21/6445 , G01N21/6452
摘要： A method and kit for determining the quantity of an analyte include providing a functionalized substrate and a reagent. The functionalized substrate includes metallic nanoparticles and a plurality of substantively identical bioactive target molecules affixed to a substrate. The bioactive target molecule binds to a particular analyte. The reagent includes identical detection molecules. Each detection molecule includes a fluorophore, and binds to a particular analyte or competes with a particular analyte for binding to the target molecule. The functionalized substrate is contacted to a test sample and the reagent. The functionalized substrate and a covering solution are exposed to polarized electromagnetic waves that excite the fluorophore. A quantity of the particular analyte in the test sample is determined based on measuring polarization anisotropy of fluorescent emissions from the substrate and the covering solution.
摘要翻译：用于确定分析物的量的方法和试剂盒包括提供官能化的底物和试剂。功能化底物包括金属纳米颗粒和固定到基底上的多个基本相同的生物活性靶分子。生物活性靶分子与特定分析物结合。该试剂包括相同的检测分子。每个检测分子包括荧光团，并结合特定分析物或与特定分析物竞争结合靶分子。将功能化底物与测试样品和试剂接触。将功能化基材和覆盖溶液暴露于激发荧光团的极化电磁波。基于测量来自基底和覆盖溶液的荧光发射的偏振各向异性来测定测试样品中特定分析物的量。

10. 发明授权

US06806089B1 Low frequency modulation sensors using nanosecond fluorophores 失效
标题翻译：低频调制传感器使用纳秒级荧光团
公开(公告)号：US06806089B1
公开(公告)日：2004-10-19
申请号：US09786627
申请日：2001-04-17
申请人： Joseph R. Lakowicz , Ignacy Gryczynski
发明人： Joseph R. Lakowicz , Ignacy Gryczynski
IPC分类号： G01N2164
CPC分类号： G01N21/6408 , G01N21/6428 , G01N2021/6415 , Y10T436/144444
摘要： Described is a new approach to fluorescence sensing based on a mixture of fluorophores, one of which is sensitive to the desired analyte. If a long lifetime analyte-insensitive fluorophore is mixed with a short lifetime analyte-sensitive fluorophore, the modulation of the emission at conveniently low frequencies becomes equal to the fractional fluorescence intensity of the sensing fluorophore. Under these conditions the modulation can be used to determine the analyte concentration. This can be used with any fluorophore which changes intensity in response to analyte, and does not require the sensing fluorophore to display a change in lifetime. The feasibility of modulation-based sensing was demonstrated using mixtures of 6-carboxyfluorescein and [Ru2,2′(bipyridyl)3]2+ as a pH sensor and of the calcium probe Fluo-3 and [Ru2,2′(bipyridyl)3]2+ as a calcium sensor.
摘要翻译：描述了基于荧光团的混合物的荧光感测的新方法，其中一种对所需分析物敏感。如果长寿命分析物不敏感的荧光团与短寿命分析物敏感的荧光团混合，则在方便的低频率下的发射调制变得等于感测荧光团的分数荧光强度。在这些条件下，调节可用于确定分析物浓度。这可以与响应于分析物改变强度的任何荧光团一起使用，并且不需要感测荧光团显示寿命的变化。使用6-羧基荧光素和[Ru2,2'（联吡啶基）3] 2+作为pH传感器和钙探针Fluo-3和[Ru2,2'（联吡啶基）的混合物来证明基于调制的感测的可行性）3] 2+作为钙传感器。

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