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    • 2. 发明授权
    • Low frequency modulation sensors using nanosecond fluorophores
    • 低频调制传感器使用纳秒级荧光团
    • US06806089B1
    • 2004-10-19
    • US09786627
    • 2001-04-17
    • Joseph R. LakowiczIgnacy Gryczynski
    • Joseph R. LakowiczIgnacy Gryczynski
    • G01N2164
    • G01N21/6408G01N21/6428G01N2021/6415Y10T436/144444
    • 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.
    • 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。
    • 3. 发明授权
    • Luminescence spectral properties of CdS nanoparticles
    • CdS纳米粒子的发光光谱性质
    • US06660379B1
    • 2003-12-09
    • US09890042
    • 2001-07-25
    • Joseph R. LakowiczIgnacy GryczynskiZygmunt Gryczynski
    • Joseph R. LakowiczIgnacy GryczynskiZygmunt Gryczynski
    • B32B516
    • G01N33/588B82Y15/00C09K11/025C09K11/565C12Q1/6816Y10T428/2982Y10T428/2991Y10T428/2998
    • 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.
    • 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。
    • 7. 发明申请
    • Apparatus and methods for surface plasmon-coupled directional emission
    • 表面等离子体耦合定向发射的装置和方法
    • US20050053974A1
    • 2005-03-10
    • US10849465
    • 2004-05-20
    • Joseph LakowiczJoanna MalickaIgnacy GryczynskiZygmunt Gryczynski
    • Joseph LakowiczJoanna MalickaIgnacy GryczynskiZygmunt Gryczynski
    • C12Q1/68G01N21/55G01N21/64G01N21/76
    • G01N21/6428G01N21/4788G01N21/6452G01N21/648G01N21/76G01N33/54373G01N2021/6432
    • 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.
    • 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。
    • 9. 发明授权
    • Surface plasmon assisted microscope
    • 表面等离子体辅助显微镜
    • US07956989B2
    • 2011-06-07
    • US12018107
    • 2008-01-22
    • Zygmunt GryczynskiIgnacy GryczynskiNils CalanderJulian Borejdo
    • Zygmunt GryczynskiIgnacy GryczynskiNils CalanderJulian Borejdo
    • G01N1/00
    • G01Q60/20G01N21/6428G01N21/6458G01N21/648G01N2021/6482G02B21/0076
    • 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.
    • 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。