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1. 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。

2. 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。

3. 发明授权

US06660379B1 Luminescence spectral properties of CdS nanoparticles 失效
标题翻译： CdS纳米粒子的发光光谱性质
公开(公告)号：US06660379B1
公开(公告)日：2003-12-09
申请号：US09890042
申请日：2001-07-25
申请人： Joseph R. Lakowicz , Ignacy Gryczynski , Zygmunt Gryczynski
发明人： Joseph R. Lakowicz , Ignacy Gryczynski , Zygmunt Gryczynski
IPC分类号： B32B516
CPC分类号： G01N33/588 , B82Y15/00 , C09K11/025 , C09K11/565 , C12Q1/6816 , Y10T428/2982 , Y10T428/2991 , Y10T428/2998
摘要： The steady state and time resolved luminescence spectral properties of two types of novel CdS nanoparticles and nanoparticles are described. CdS nanoparticles formed in the presence of an amine-terminated dendrimer show blue emission. The emission wavelength of these nanoparticles depended on the excitation wavelength. The CdS/dendrimer nanoparticles display polarized emission with the anisotropy rising progressively from 340 to 420 nm excitation, reaching a maximal anisotropy value in excess of 0.3. A new constant positive polarized emission from luminescent nanoparticles is also described. Polyphosphate-stabilized CdS nanoparticles are described that display a longer wavelength red emission maximum than bulk CdS and display a zero anisotropy for all excitation wavelengths. Both nanoparticles display strongly heterogeneous intensity decays with mean decay times of 93 ns and 10 &mgr;s for the blue and red emitting particles, respectively. Both types of nanoparticles were several times more photostable upon continous illumination than fluorescein. In spite of the long decay times the nanoparticles are mostly insensitive to dissolved oxygen but are quenched by iodide. These nanoparticles can provide a new class of luminophores for use in chemical sensing, DNA sequencing, high throughput screening and other applications.
摘要翻译：描述了两种新型CdS纳米颗粒和纳米颗粒的稳态和时间分辨发光光谱性质。在胺封端的树枝状大分子存在下形成的CdS纳米颗粒显示蓝色发射。这些纳米颗粒的发射波长取决于激发波长。 CdS /树枝状大分子纳米粒子显示极化发射，各向异性从340到420nm激发逐渐上升，达到超过0.3的最大各向异性值。还描述了来自发光纳米颗粒的新的恒定的正极化发射。描述了多磷酸盐稳定的CdS纳米颗粒，其显示比体积CdS更长的波长红色发射，并且对于所有激发波长显示零各向异性。两种纳米颗粒都显示强烈的非均匀强度衰变，其平均衰减时间分别为93ns和10μs，分别为蓝色和红色发射粒子。两种类型的纳米颗粒在连续照射下比荧光素多几倍的光稳定性。尽管长时间的衰变时间，纳米颗粒对溶解氧大多不敏感，但被碘化物淬灭。这些纳米颗粒可以提供一类新的发光体，用于化学感测，DNA测序，高通量筛选和其他应用。

4. 发明授权

US06395556B1 Polarization based sensing 失效
标题翻译：基于极化的感测
公开(公告)号：US06395556B1
公开(公告)日：2002-05-28
申请号：US09831625
申请日：2001-08-07
申请人： Joseph R. Lakowicz , Ignacy Gryczynski , Zygmunt Gryczynski
发明人： Joseph R. Lakowicz , Ignacy Gryczynski , Zygmunt Gryczynski
IPC分类号： G01N2164
CPC分类号： G01N21/6445 , G01N21/6428 , G01N2021/6441 , Y10T436/144444 , Y10T436/209163
摘要： The present invention relates to the determination of the presence or concentration of an analyte in a sample by visual or electronic element, using polarization based sensing techniques (14) employing fluorescent sensing (11) and reference molecules (10).
摘要翻译：本发明涉及使用采用荧光检测（11）和参考分子（10）的基于偏振的感测技术（14），通过视觉或电子元件确定样品中分析物的存在或浓度。

5. 发明授权

US5196709A Fluorometry method and apparatus using a semiconductor laser diode as a light source 失效
标题翻译：使用半导体激光二极管作为光源的荧光测定法和装置
公开(公告)号：US5196709A
公开(公告)日：1993-03-23
申请号：US695074
申请日：1991-05-03
申请人： Klaus W. Berndt , Ignacy Gryczynski , Joseph R. Lakowicz
发明人： Klaus W. Berndt , Ignacy Gryczynski , Joseph R. Lakowicz
IPC分类号： G01N21/64
CPC分类号： G01N21/6408 , G01N2201/0612 , G01N2201/0691
摘要： A method and apparatus for detecting the change in phase angle and/or modulation of emitted fluorescence of a fluorophore excited by modulated light from a laser diode. The light is both monochromatic and coherent, and can contain harmonic frequency components. The laser diode can be used in frequency-domain or phase-modulation measurements of fluorescence, using either phase or modulation techniques. The present invention is an improvement of the prior use of a frequency-doubled, cavity-dumped, dye laser in fluorometers. The invention provides an inexpensive light excitation source that is small in size, easily manageable, allows for short measurement times, and has lower power requirements.
摘要翻译：一种用于检测由激光二极管调制的光激发的荧光团的相位角变化和/或发射荧光调制的方法和装置。光是单色和相干的，并且可以包含谐波频率分量。使用相位或调制技术，激光二极管可用于荧光的频域或相位调制测量。本发明是在荧光计中先前使用倍频，空腔倾倒的染料激光器的改进。本发明提供了一种廉价的光激发源，其尺寸小，易于管理，允许短的测量时间，并且具有较低的功率要求。

6. 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。

7. 发明授权

US06432637B1 Method for determining a base sequence of a nucleotide strand 失效
标题翻译：确定核苷酸链的碱基序列的方法
公开(公告)号：US06432637B1
公开(公告)日：2002-08-13
申请号：US08990539
申请日：1997-12-15
申请人： Joseph R. Lakowicz
发明人： Joseph R. Lakowicz
IPC分类号： C12Q168
CPC分类号： C12Q1/6869 , C07H19/14 , C07H21/00 , C12Q2563/137 , C12Q2535/125 , C12Q2535/101
摘要： A method for determining a base sequence of a nucleotide strand in a sample utilizes a probe including a fluorescent metal-ligand complex coupled to a first oligonucleotide having a sequence complementary to the first fragment of the strand. The first mixture is exposed to an exciting amount of radiation, and the fluorescence of the metal-ligand complex is detected. The first base sequence of the first fragment is identified based on fluorescence of the metal ligand complex. A second probe differing from the first by at least one base is provided. A second base of the second fragment is identified based on the fluorescence of the metal-ligand complex of the second probe. The second base sequence is compared to the first base sequence to identify a difference between the first and second sequences to determine a base sequence of the nucleotide strand.
摘要翻译：用于测定样品中核苷酸链的碱基序列的方法利用包含与具有与该链的第一片段互补的序列的第一寡核苷酸偶联的荧光金属 - 配体复合物的探针。将第一混合物暴露于激发量的辐射，并检测金属 - 配体络合物的荧光。基于金属配体络合物的荧光来鉴定第一片段的第一碱基序列。提供了与第一至少一个基座不同的第二探针。基于第二探针的金属 - 配体复合物的荧光来鉴定第二片段的第二碱基。将第二碱基序列与第一碱基序列进行比较以鉴定第一和第二序列之间的差异，以确定核苷酸链的碱基序列。

8. 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。

9. 发明授权

US5504337A Method and apparatus for performing phase fluorescence lifetime measurements in flow cytometry 失效
标题翻译：用于在流式细胞仪中进行相位荧光寿命测量的方法和装置
公开(公告)号：US5504337A
公开(公告)日：1996-04-02
申请号：US50122
申请日：1993-04-12
申请人： Joseph R. Lakowicz , Klaus W. Berndt , Robert A. Hoffman , Bertram G. Pinsky
发明人： Joseph R. Lakowicz , Klaus W. Berndt , Robert A. Hoffman , Bertram G. Pinsky
IPC分类号： G01N15/14 , G01N21/64
CPC分类号： G01N15/1434 , G01N21/6408 , G01N15/1459 , G01N2015/1454
摘要： A method and apparatus for identifying individual particles or cells which have been labeled with different fluorochromes, on the basis of the lifetime of their fluorescence, or based on different decay times for a fluorochrome in different cells.
摘要翻译： PCT No.PCT / US91 / 07259 Sec。 371日期：1993年4月12日 102（e）日期1993年4月12日PCT 1991年10月10日PCT PCT。公开号WO92 / 07245 日期：1992年4月30日。一种用于识别已经用不同荧光染料标记的单个颗粒或细胞的方法和装置，基于其荧光的寿命，或者基于不同细胞中的荧光染料的不同衰减时间。

10. 发明授权

US5409835A Long-wavelength fluorescent probe compounds for calcium ions and their use in ratiometrically measuring calcium ion concentrations 失效
标题翻译：用于钙离子的长波长荧光探针化合物及其在比例测量钙离子浓度中的用途
公开(公告)号：US5409835A
公开(公告)日：1995-04-25
申请号：US998849
申请日：1992-12-30
申请人： Joseph R. Lakowicz , Engin U. Akkaya
发明人： Joseph R. Lakowicz , Engin U. Akkaya
IPC分类号： C07D277/64 , G01N33/52 , G01N33/58 , G01N33/84 , C07D277/62 , C07D277/70 , C07D335/04
CPC分类号： G01N33/582 , C07D277/64 , G01N33/52 , G01N33/84 , Y10S436/80
摘要： Fluorescent, calcium-binding probe compounds having visible-light excitation and emission wavelengths, and a method of usage for these probe compounds in making wavelength-ratiometric or intensity-ratiometric measurements of calcium ion concentration in samples. The probe compounds generally relate to a heterocyclic part as bonded to a Ca.sup.2+ binding unit, namely, BAPTA, via an ethylenically unsaturated group; preferred examples of the heterocyclic group include benzothiazole, naphthothiazole, thiaflavin, indolenine, chloroindolenine, methoxybenzothiazole, and methoxyindolenine.
摘要翻译：具有可见光激发和发射波长的荧光，钙结合探针化合物以及这些探针化合物在制备样品中钙离子浓度的波长比例或强度比例测量时的使用方法。探针化合物通常涉及通过烯属不饱和基团键合到Ca 2+结合单元即BAPTA的杂环部分; 杂环基的优选实例包括苯并噻唑，萘并噻唑，硫黄素，吲哚宁，氯代二甲苯胺，甲氧基苯并噻唑和甲氧基二氢化茚。

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