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1. 发明申请

US20090218516A1 Ratiometric Surface Plasmon Coupled Emission Detector 有权
标题翻译：比例表面等离子体耦合发射检测器
公开(公告)号：US20090218516A1
公开(公告)日：2009-09-03
申请号：US12356288
申请日：2009-01-20
申请人： Zygmunt Gryczynski , Ignacy Gryczynski , Evgenia Matveeva , Julian Borejdo
发明人： Zygmunt Gryczynski , Ignacy Gryczynski , Evgenia Matveeva , Julian Borejdo
IPC分类号： G01N21/64
CPC分类号： G01N21/648 , G01N21/553
摘要： The present invention includes methods for ratiometric detection of analytes by surface plasmon coupled emission detection that includes disposing a target on the metal layer of a surface plasmon resonance detection system; coupling a first analyte to a first fluorescent dye and a second analyte to a second fluorescent dye; contacting the first and second analytes to the target on the surface plasmon resonance detection system; and measuring the intensity of a first and a second surface plasmon resonance enhanced fluorescence emission ring, wherein the first and second rings, respectively, quantitatively represents the amount of first and second analyte within 50 nanometers of the metal surface.
摘要翻译：本发明包括通过表面等离子体激元耦合发射检测对分析物进行比例检测的方法，其包括在表面等离子体共振检测系统的金属层上设置目标物; 将第一分析物与第一荧光染料和第二分析物耦合到第二荧光染料; 使第一和第二分析物与表面等离子体共振检测系统上的靶接触; 以及测量第一和第二表面等离子体共振增强荧光发射环的强度，其中第一和第二环分别定量地表示在金属表面的50纳米内的第一和第二分析物的量。

2. 发明申请

US20050053974A1 Apparatus and methods for surface plasmon-coupled directional emission 审中-公开
标题翻译：表面等离子体耦合定向发射的装置和方法
公开(公告)号：US20050053974A1
公开(公告)日：2005-03-10
申请号：US10849465
申请日：2004-05-20
申请人： Joseph Lakowicz , Joanna Malicka , Ignacy Gryczynski , Zygmunt Gryczynski
发明人： Joseph Lakowicz , Joanna Malicka , Ignacy Gryczynski , Zygmunt Gryczynski
IPC分类号： C12Q1/68 , G01N21/55 , G01N21/64 , G01N21/76
CPC分类号： G01N21/6428 , G01N21/4788 , G01N21/6452 , G01N21/648 , G01N21/76 , G01N33/54373 , G01N2021/6432
摘要： Methods and apparatus for fluorescence detection which can increase sensitivity by as much as 20 to 1000-fold are described. This method can preferably also decrease the contribution of sample autofluorescence to the detected signal. The method uses coupling of excited fluorophores with the surface plasmon resonance present in thin conductive films, for example silver, gold, aluminum, copper, or the like. The phenomenon of surface plasmon-coupled emission (SPCE) occurs for fluorophores in a volume adjacent to the conductive layer. This interaction is independent of the mode of excitation, that is, does not require evanescent wave or surface-plasmon excitation. However, such modes of excitation can be advantageous. SPCE can occur over a narrow angular distribution, converting normally isotropic emission into easily collected directional emission. In preferred embodiments, up to 50% of the emission from unoriented samples can be collected, usually much more than typical fluorescence collection efficiencies, which can be 1% or less. Examples are presented showing how simple optical configurations can be used in diagnostics, sensing, or biotechnology applications. Surface plasmon-coupled emission is likely to find widespread applications throughout the biosciences.
摘要翻译：描述了将灵敏度提高多达20到1000倍的用于荧光检测的方法和装置。该方法也可以优选地降低样品自发荧光对检测信号的贡献。该方法使用激发的荧光团与存在于薄导电膜（例如银，金，铝，铜等）中的表面等离子体共振的耦合。在与导电层相邻的体积中的荧光团发生表面等离激元耦合发射（SPCE）的现象。这种相互作用与激发模式无关，也就是说，不需要ev逝波或表面等离子体激发。然而，这种激发模式可能是有利的。 SPCE可以发生在窄角分布上，将正常的各向同性发射转换成易于收集的定向发射。在优选的实施方案中，可以收集来自无取向样品的高达50％的发射，通常比典型的荧光收集效率高得多，其可以为1％或更少。现在举例说明如何在诊断，感测或生物技术应用中使用简单的光学配置。表面等离子体耦合发射很可能在整个生物科学领域得到广泛的应用。

3. 发明申请

US20110207614A1 Polarized Flourescent Nanospheres 有权
标题翻译：极化荧光纳米球
公开(公告)号：US20110207614A1
公开(公告)日：2011-08-25
申请号：US13030048
申请日：2011-02-17
申请人： Zygmunt Gryczynski , Ignacy Gryczynski , Rafal Luchowski , Julian Borejdo
发明人： Zygmunt Gryczynski , Ignacy Gryczynski , Rafal Luchowski , Julian Borejdo
IPC分类号： C40B30/00 , C12Q1/02 , C09K11/06 , C09K11/02 , H01F1/00 , H01L31/00 , B82Y15/00
CPC分类号： B82Y15/00 , B82Y20/00 , B82Y25/00 , C09K11/02 , C09K11/06 , H01F1/0063
摘要： The present invention provides partially fluorescent nanoparticles and methods of making and using the partially fluorescent nanoparticle having a nanoparticle with a matrix and a fluorescent dye dispersed in or about the matrix, wherein at least a portion of the fluorescent dye has been anisotropically bleached.
摘要翻译：本发明提供部分荧光纳米颗粒和制备和使用具有分散在基质中或其周围的基质和荧光染料的纳米颗粒的部分荧光纳米颗粒的方法，其中至少一部分荧光染料已被各向异性漂白。

4. 发明授权

US07956989B2 Surface plasmon assisted microscope 有权
标题翻译：表面等离子体辅助显微镜
公开(公告)号：US07956989B2
公开(公告)日：2011-06-07
申请号：US12018107
申请日：2008-01-22
申请人： Zygmunt Gryczynski , Ignacy Gryczynski , Nils Calander , Julian Borejdo
发明人： Zygmunt Gryczynski , Ignacy Gryczynski , Nils Calander , Julian Borejdo
IPC分类号： G01N1/00
CPC分类号： G01Q60/20 , G01N21/6428 , G01N21/6458 , G01N21/648 , G01N2021/6482 , G02B21/0076
摘要： The present invention includes a microscope and a method for using the microscope for single molecule with reduced photobleaching of a fluorophore (20) that includes a light translucent material (16); a metal layer (18) disposed on the light translucent material (16); a medium (15) disposed on the metal layer (18), the medium (15) having one or more fluorophores (20) capable of binding a target analyte (e.g., inside a cell); a microscope positioned to observe the surface plasmon emissions from the one or more fluorophores (20) within 50 nanometers of the surface of the metal layer (18); an excitation source capable of exciting the one or more fluorophores (20), the excitation source positioned to strike the light translucent material (16) at a first angle; and a light detector (38) that selectively detects emitted light generated by excited fluorophores (20) at a second angle (22), wherein light emitted by the one or more fluorophores (20) at the surface plasmon angle is detected through the microscope, such that single molecules may be detected without significantly degrading fluorophore (20) emissions.
摘要翻译：本发明包括一种显微镜和一种使用该显微镜用于具有减少的光漂白的单分子的显微镜，所述荧光团（20）包括光半透明材料（16）; 设置在所述光半透明材料（16）上的金属层（18）; 设置在所述金属层（18）上的介质（15），所述介质（15）具有一个或多个能够结合目标分析物（例如细胞内）的荧光团（20）; 位于观察来自金属层（18）的表面的50纳米内的一个或多个荧光团（20）的表面等离子体激元发射的显微镜; 激励源，其能够激发所述一个或多个荧光团（20），所述激发源被定位成以第一角度撞击所述光半透明材料（16）; 以及光检测器（38），其以第二角度（22）选择性地检测由激发的荧光团（20）产生的发射光，其中通过显微镜检测由所述一个或多个荧光团（20）以表面等离子体角度发射的光，使得可以在不显着降解荧光团（20）排放的情况下检测单分子。

5. 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。

6. 发明授权

US08159676B2 Ratiometric surface plasmon coupled emission detector 有权
标题翻译：比例表面等离子体耦合发射检测器
公开(公告)号：US08159676B2
公开(公告)日：2012-04-17
申请号：US12356288
申请日：2009-01-20
申请人： Zygmunt Gryczynski , Ignacy Gryczynski , Evgenia Matveeva , Julian Borejdo
发明人： Zygmunt Gryczynski , Ignacy Gryczynski , Evgenia Matveeva , Julian Borejdo
IPC分类号： G01N21/55
CPC分类号： G01N21/648 , G01N21/553
摘要： The present invention includes methods for ratiometric detection of analytes by surface plasmon coupled emission detection that includes disposing a target on the metal layer of a surface plasmon resonance detection system; coupling a first analyte to a first fluorescent dye and a second analyte to a second fluorescent dye; contacting the first and second analytes to the target on the surface plasmon resonance detection system; and measuring the intensity of a first and a second surface plasmon resonance enhanced fluorescence emission ring, wherein the first and second rings, respectively, quantitatively represents the amount of first and second analyte within 50 nanometers of the metal surface.
摘要翻译：本发明包括通过表面等离子体激元耦合发射检测对分析物进行比例检测的方法，其包括在表面等离子体共振检测系统的金属层上设置目标物; 将第一分析物与第一荧光染料和第二分析物耦合到第二荧光染料; 使第一和第二分析物与表面等离子体共振检测系统上的靶接触; 以及测量第一和第二表面等离子体共振增强荧光发射环的强度，其中第一和第二环分别定量地表示在金属表面的50纳米内的第一和第二分析物的量。

7. 发明授权

US5759767A Two-photon and multi-photon measurement of analytes in animal and human tissues and fluids 失效
标题翻译：动物和人体组织和液体中分析物的双光子和多光子测量
公开(公告)号：US5759767A
公开(公告)日：1998-06-02
申请号：US731270
申请日：1996-10-11
申请人： Joseph R. Lakowicz , Thomas G. Burke , Ignacy Gryczynski , Henryk Malak
发明人： Joseph R. Lakowicz , Thomas G. Burke , Ignacy Gryczynski , Henryk Malak
IPC分类号： A61K41/00 , G01N33/487 , C12Q1/00 , G01N33/53 , G01N33/567
CPC分类号： A61K41/008 , A61N5/062 , Y10S435/968 , Y10S436/80
摘要： A method of measuring an analyte present in animal (e.g., human) tissue or fluids such as blood or plasma. The analyte is multi-photon excitable (e.g., two-photon excitable) at a first wavelength at which the animal tissue is substantially non-absorbing. The analyte fluoresces at a second wavelength upon being excited at the first wavelength. The animal tissue is irradiated with radiation at the first wavelength so as to excite the analyte through absorption by the analyte of two or more photons of the radiation at the first wavelength. Excitation of the analyte results in a fluorescent emission from the analyte of radiation at the second wavelength. The emission at the second wavelength is detected, and the concentration of analyte determined based on the detected emission.
摘要翻译：测量存在于动物（例如人）组织或诸如血液或血浆的流体中的分析物的方法。分析物是动物组织基本上不吸收的第一波长处的多光子可激发（例如，双光子可激发的）。当第一波长被激发时，分析物以第二波长发荧光。用第一波长的辐射照射动物组织，以便通过被第一波长的辐射的两个或多个光子的分析物吸收来激发分析物。分析物的激发导致来自第二波长的辐射分析物的荧光发射。检测第二波长的发射，并且基于检测到的发射确定分析物的浓度。

8. 发明授权

US09376315B2 Polarized fluorescent nanospheres 有权
标题翻译：极化荧光纳米球
公开(公告)号：US09376315B2
公开(公告)日：2016-06-28
申请号：US13030048
申请日：2011-02-17
申请人： Zygmunt Gryczynski , Ignacy Gryczynski , Rafal Luchowski , Julian Borejdo
发明人： Zygmunt Gryczynski , Ignacy Gryczynski , Rafal Luchowski , Julian Borejdo
IPC分类号： C09K11/06 , B82Y20/00 , B82Y5/00 , B82Y15/00 , B82Y25/00 , C09K11/02 , H01F1/00
CPC分类号： B82Y15/00 , B82Y20/00 , B82Y25/00 , C09K11/02 , C09K11/06 , H01F1/0063
摘要： The present invention provides partially fluorescent nanoparticles and methods of making and using the partially fluorescent nanoparticle having a nanoparticle with a matrix and a fluorescent dye dispersed in or about the matrix, wherein at least a portion of the fluorescent dye has been anisotropically bleached.
摘要翻译：本发明提供部分荧光纳米颗粒和制备和使用具有分散在基质中或其周围的基质和荧光染料的纳米颗粒的部分荧光纳米颗粒的方法，其中至少一部分荧光染料已经各向异性漂白。

9. 发明申请

US20110089317A1 Polarization Standards for Microscopy 审中-公开
标题翻译：显微镜极化标准
公开(公告)号：US20110089317A1
公开(公告)日：2011-04-21
申请号：US12909628
申请日：2010-10-21
申请人： Ignacy Gryczynski , Zygmunt Gryczynski , Rafal Luchowski , Julian Borejdo
发明人： Ignacy Gryczynski , Zygmunt Gryczynski , Rafal Luchowski , Julian Borejdo
IPC分类号： G01D18/00 , G01N31/00 , C09K11/06
CPC分类号： G01J3/28 , A61B5/0075 , A61B2560/0233 , G01J3/42 , G01J3/447 , G01N21/21 , G01N21/278 , G01N21/6445 , G01N21/6458 , G01N2021/6417
摘要： The present invention describes the development of thin film calibration strips for microscopy/spectroscopy systems and a simple method/routine to conduct instrument calibration using partially (uniaxially) oriented strip to calibrate microscopy system without the prior knowledge of exact polarization of the strip. The invention describes results from studies including a styryl derivative (LDS 798) embedded in poly(vinyl alcohol) (PVA) film. These films were progressively stretched up to 8 folds. Vertical and horizontal components of absorptions and fluorescence were measured and dichroic ratios were determined for different film stretching ratios. The stretched films have high polarization values for isotropic excitation. The isotropic and stretched PVA films doped with LDS 798 can be used as etalons in near infra red (NIR) spectroscopic measurements. The high polarization standards of the present invention have applications in NIR imaging microscopy where they can be used for correcting for instrumental factor in polarization measurements.
摘要翻译：本发明描述了用于显微镜/光谱系统的薄膜校准条的开发以及使用部分（单轴）取向的条进行仪器校准的简单方法/程序，以校准显微镜系统，而无需预先知道条的精确极化。本发明描述了嵌入聚（乙烯醇）（PVA）膜中的苯乙烯基衍生物（LDS 798）的研究结果。这些胶片逐渐拉伸至8倍。测量吸收和荧光的垂直和水平分量，并确定不同膜拉伸比的二向色比。拉伸膜对各向同性激发具有高极化值。掺杂LDS 798的各向同性和拉伸PVA膜可用于近红外（NIR）光谱测量中的标准具。本发明的高极化标准在NIR成像显微镜中具有应用，可用于校正极化测量中的仪器因素。

10. 发明申请

US20080231834A1 SURFACE PLASMON ASSISTED MICROSCOPE 有权
标题翻译：表面等离子体辅助显微镜
公开(公告)号：US20080231834A1
公开(公告)日：2008-09-25
申请号：US12018107
申请日：2008-01-22
申请人： Zygmunt Gryczynski , Ignacy Gryczynski , Nils Calander , Julian Borejdo
发明人： Zygmunt Gryczynski , Ignacy Gryczynski , Nils Calander , Julian Borejdo
IPC分类号： G01N1/00
CPC分类号： G01Q60/20 , G01N21/6428 , G01N21/6458 , G01N21/648 , G01N2021/6482 , G02B21/0076
摘要： The present invention includes a microscope and a method for using the microscope for single molecule with reduced photobleaching of a fluorophore (20) that includes a light translucent material (16); a metal layer (18) disposed on the light translucent material (16); a medium (15) disposed on the metal layer (18), the medium (15) having one or more fluorophores (20) capable of binding a target analyte (e.g., inside a cell); a microscope positioned to observe the surface plasmon emissions from the one or more fluorophores (20) within 50 nanometers of the surface of the metal layer (18); an excitation source capable of exciting the one or more fluorophores (20), the excitation source positioned to strike the light translucent material (16) at a first angle; and a light detector (38) that selectively detects emitted light generated by excited fluorophores (20) at a second angle (22), wherein light emitted by the one or more fluorophores (20) at the surface plasmon angle is detected through the microscope, such that single molecules may be detected without significantly degrading fluorophore (20) emissions.
摘要翻译：本发明包括一种显微镜和一种使用该显微镜用于具有减少的光漂白的单分子的显微镜，所述荧光团（20）包括光半透明材料（16）; 设置在所述光半透明材料（16）上的金属层（18）; 设置在所述金属层（18）上的介质（15），所述介质（15）具有一个或多个能够结合目标分析物（例如细胞内）的荧光团（20）; 位于观察来自金属层（18）的表面的50纳米内的一个或多个荧光团（20）的表面等离子体激元发射的显微镜; 激励源，其能够激发所述一个或多个荧光团（20），所述激发源被定位成以第一角度撞击所述光半透明材料（16）; 以及光检测器（38），其以第二角度（22）选择性地检测由激发的荧光团（20）产生的发射光，其中通过显微镜检测由所述一个或多个荧光团（20）以表面等离子体角度发射的光，使得可以在不显着降解荧光团（20）排放的情况下检测单分子。

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