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1. 发明申请

US20120276578A1 Phasor Method to Fluorescence Lifetime Microscopy to Discriminate Metabolic State of Cells in Living Tissue 审中-公开
标题翻译：荧光终生显微镜的相量方法来鉴别生活组织中细胞的代谢状态
公开(公告)号：US20120276578A1
公开(公告)日：2012-11-01
申请号：US13283356
申请日：2011-10-27
申请人： Chiara Stringari , Enrico Gratton , Michelle Digman , Peter Donovan
发明人： Chiara Stringari , Enrico Gratton , Michelle Digman , Peter Donovan
IPC分类号： G01N21/64 , C12M1/34
CPC分类号： G01N21/6486 , G01N21/6408 , G01N21/6458
摘要： “A label-free imaging method to monitor stem cell metabolism discriminates different states of stem cell as they differentiate in a living tissues. We use intrinsic fluorescence biomarkers and the phasor approach to Fluorescence Lifetime Imaging Microscopy (FLIM). We identify and map intrinsic fluorophores such as collagen, retinol, retinoic acid, flavins, nicotinamide adenine dinucleotide (NADH) and porphyrin. We measure the phasor values of germ cells in C. Elegans germ line. Their metabolic fingerprint cluster according to their differentiation state, reflecting changes in FAD concentration and NADH binding during the differentiation pathway. The phasor approach to lifetime imaging provides a label-free, fit-free and sensitive method to identify different metabolic state of cells during differentiation, to sense small changes in the redox state of cells and may identify symmetric and asymmetric divisions and predict cell fate.”
摘要翻译：监测干细胞代谢的无标记成像方法会区分干细胞在活组织中分化的不同状态。我们使用内在荧光生物标志物和荧光寿命成像显微镜（FLIM）的相量方法。我们识别和映射内在的荧光团，如胶原，视黄醇，视黄酸，黄素，烟酰胺腺嘌呤二核苷酸（NADH）和卟啉。我们测量C. Elegans种系中生殖细胞的相量值。他们的代谢指纹簇根据其分化状态，反映分化途径中FAD浓度和NADH结合的变化。终生成像的相量方法提供了无标记，无配对和敏感的方法，以鉴别细胞在分化期间的不同代谢状态，以感测细胞氧化还原状态的小变化，并且可以鉴定对称和不对称分裂并预测细胞命运。

2. 发明授权

US10222335B2 Phasor method to fluorescence lifetime microscopy to discriminate metabolic state of cells in living tissue 有权
公开(公告)号：US10222335B2
公开(公告)日：2019-03-05
申请号：US13283356
申请日：2011-10-27
申请人： Chiara Stringari , Enrico Gratton , Michelle Digman , Peter Donovan
发明人： Chiara Stringari , Enrico Gratton , Michelle Digman , Peter Donovan
IPC分类号： G01N21/64
摘要： A label-free imaging method to monitor stem cell metabolism discriminates different states of stem cell as they differentiate in a living tissues. We use intrinsic fluorescence biomarkers and the phasor approach to Fluorescence Lifetime Imaging Microscopy (FLIM). We identify and map intrinsic fluorophores such as collagen, retinol, retinoic acid, flavins, nicotinamide adenine dinucleotide (NADH) and porphyrin. We measure the phasor values of germ cells in C. Elegans germ line. Their metabolic fingerprint cluster according to their differentiation state, reflecting changes in FAD concentration and NADH binding during the differentiation pathway. The phasor approach to lifetime imaging provides a label-free, fit-free and sensitive method to identify different metabolic state of cells during differentiation, to sense small changes in the redox state of cells and may identify symmetric and asymmetric divisions and predict cell fate.

3. 发明授权

US09110282B2 Nanometer-scale optical imaging by the modulation tracking (MT) method 有权
标题翻译：通过调制跟踪（MT）方法进行纳米级光学成像
公开(公告)号：US09110282B2
公开(公告)日：2015-08-18
申请号：US13435387
申请日：2012-03-30
申请人： Luca Lanzano , Michelle Digman , Enrico Gratton
发明人： Luca Lanzano , Michelle Digman , Enrico Gratton
IPC分类号： G01C3/08 , G02B21/00 , G01B9/04 , G01N21/64 , B82Y35/00
CPC分类号： G02B21/0012 , B82Y35/00 , G01B9/04 , G01N21/6458 , G02B2207/113
摘要： An optical imaging method based on a feedback principle in which the specific scan pattern is adapted according to the shape of the sample. The feedback approach produces nanometer-resolved three dimensional images of very small and moving features in live cells and in a matter of seconds. Images of microvilli in live cultured opossum kidney cells expressing NaPi co-transporter proteins with different GFP constructs and images of cell protrusions in a collagen matrix are produced with a resolution of about 20 nm. Along cell protrusions in three dimensional cellular adhesions could be identified to the extracellular matrix.
摘要翻译：基于反馈原理的光学成像方法，其中特定扫描图案根据样品的形状进行调整。反馈方法在活细胞中产生非常小且移动的特征的纳米解析的三维图像，并且在几秒钟内。用活性培养的负鼠肾细胞中表达具有不同GFP构建体的NaPi共转运蛋白和胶原基质中细胞突起的图像的微绒毛的图像以约20nm的分辨率产生。沿着三维细胞粘连中的细胞突起可以被鉴定为细胞外基质。

4. 发明申请

US20120250000A1 NANOMETER-SCALE OPTICAL IMAGING BY THE MODULATION TRACKING (MT) METHOD 有权
标题翻译：通过调制跟踪（MT）方法进行的NANOMETER-SCALE OPTICAL IMAGING
公开(公告)号：US20120250000A1
公开(公告)日：2012-10-04
申请号：US13435387
申请日：2012-03-30
申请人： Luca Lanzano , Michelle Digman , Enrico Gratton
发明人： Luca Lanzano , Michelle Digman , Enrico Gratton
IPC分类号： G01C3/08 , G01B11/24
CPC分类号： G02B21/0012 , B82Y35/00 , G01B9/04 , G01N21/6458 , G02B2207/113
摘要： An optical imaging method based on a feedback principle in which the specific scan pattern is adapted according to the shape of the sample. The feedback approach produces nanometer-resolved three dimensional images of very small and moving features in live cells and in a matter of seconds. Images of microvilli in live cultured opossum kidney cells expressing NaPi co-transporter proteins with different GFP constructs and images of cell protrusions in a collagen matrix are produced with a resolution of about 20 nm. Along cell protrusions in three dimensional cellular adhesions could be identified to the extracellular matrix.
摘要翻译：基于反馈原理的光学成像方法，其中特定扫描图案根据样品的形状进行调整。反馈方法在活细胞中产生非常小且移动的特征的纳米解析的三维图像，并且在几秒钟内。用活性培养的负鼠肾细胞中表达具有不同GFP构建体的NaPi共转运蛋白和胶原基质中细胞突起的图像的微绒毛的图像以约20nm的分辨率产生。沿着三维细胞粘连中的细胞突起可以被鉴定为细胞外基质。

5. 发明授权

US08256898B2 Method and device for tear film analysis 有权
标题翻译：用于泪膜分析的方法和装置
公开(公告)号：US08256898B2
公开(公告)日：2012-09-04
申请号：US12813423
申请日：2010-06-10
申请人： Enrico Gratton , Kaveh Azartash , Luisa Marsili
发明人： Enrico Gratton , Kaveh Azartash , Luisa Marsili
IPC分类号： A61B3/14 , A61B3/10
CPC分类号： A61B3/101
摘要： Tear film stability has an important role in the quality of vision. A system and method for performing Fluctuation Analysis of Spatial Image Correlation (FASIC) provides for a non-invasive system and method for evaluating the dynamics of the tear film surface using spatial autocorrelation analysis. With FASIC, a series of images are obtained using illumination and a camera. The spatial autocorrelation is calculated for image frames produced by the camera. A sinusoidal background appears in this correlation together with other features. The changes in the sinusoidal background of the spatial autocorrelation is extracted and monitored over time. The spatial period of this sinusoidal background correlates with the thickness of the tear film. In this regard, one is able to derive the tear film thickness from the period of this sinusoidal background.
摘要翻译：撕裂膜的稳定性在视觉质量中起着重要的作用。用于执行空间图像相关性（FASIC）的波动分析的系统和方法提供了一种用于使用空间自相关分析来评估泪膜表面的动力学的非侵入性系统和方法。使用FASIC，使用照明和相机获得一系列图像。对由相机产生的图像帧计算空间自相关。这种相关性与其他特征一起出现正弦背景。空间自相关正弦背景的变化随时间推移和监测。该正弦背景的空间周期与泪膜的厚度相关。在这方面，能够从该正弦背景的周期中导出泪膜厚度。

6. 发明授权

US5323010A Time resolved optical array detectors and CCD cameras for frequency domain fluorometry and/or phosphorimetry 失效
标题翻译：用于频域荧光测定和/或荧光测定的时间分辨光学阵列检测器和CCD照相机
公开(公告)号：US5323010A
公开(公告)日：1994-06-21
申请号：US983829
申请日：1992-12-01
申请人： Enrico Gratton , Martin VandeVen , Beniamino Barbieri
发明人： Enrico Gratton , Martin VandeVen , Beniamino Barbieri
IPC分类号： G01J3/44 , G01N21/64
CPC分类号： G01N21/6408 , G01J3/4406 , G01N2021/1772 , G01N2021/1789
摘要： An apparatus for cross-correlation frequency domain fluorometry-phosphorimetry comprises a source of electromatic radiation and means for amplitude modulating the radiation at the first frequency. The amplitude modulated radiation is directed at a sample, while an optical array detector measures the resulting luminescence of the sample. A signal is provided coherent with the amplitude modulated electromagnetic radiation signals, at a second frequency which is different from the first frequency. The apparatus has the capability for shutting off and turning on the coherent signal at the second frequency in a cycle which is at a third frequency that is different from the difference between the first and second frequencies. This produces a resultant signal at a frequency derived from the difference and the third frequency. The resultant signal, when turned on, modulates the gain of the detecting means or multiplies its output, depending upon the nature of the detecting means. The amount of luminescence measured by the optical array detector is read when the coherent signal is off in its cycle of the third frequency. A signal from the reader described above is then detected at a frequency of the resultant signal to determine phase shift and modulation changes of the luminescence.
摘要翻译：用于互相关频域荧光测定法的装置包括电离辐射源和用于以第一频率幅度调制辐射的装置。幅度调制辐射指向样本，而光学阵列检测器测量样品的所得发光。在与第一频率不同的第二频率处提供与调幅电磁辐射信号相干的信号。该装置具有在与第一和第二频率之间的差异不同的周期中以第二频率关闭和接通相干信号的能力。这产生一个从差值和第三个频率得到的频率的合成信号。所得到的信号在导通时根据检测装置的性质调制检测装置的增益或乘法器的输出。当相干信号在其第三频率的周期中关闭时，读取由光学阵列检测器测量的发光量。然后在所得信号的频率处检测来自上述读取器的信号，以确定发光的相移和调制变化。

7. 发明授权

US5212386A High speed cross-correlation frequency domain fluorometry-phosphorimetry 失效
标题翻译：高速互相关频域荧光测定法 - 荧光测定法
公开(公告)号：US5212386A
公开(公告)日：1993-05-18
申请号：US807261
申请日：1991-12-13
申请人： Enrico Gratton , Beniamino Barbieri
发明人： Enrico Gratton , Beniamino Barbieri
IPC分类号： G01N21/64
CPC分类号： G01N21/6408
摘要： Apparatus for cross-correlation frequency domain fluorometry and/or phosphorimetry in which means are provided for sequentially performing runs of the cross correlation frequency domain fluorometry and/or phosphorimetry at sequentially differing first and second frequencies. The intensities of signal responses of the runs are detected at the respective cross-correlation frequency in each run. The detection of the signal response is prolonged in each run until an integrated signal with a specified standard deviation has been acquired at each of the differing runs. Preferably the sequential runs are automatically executed by a program. Also, the waveforms sensed by deriving the resultant signal response in each run are folded. That is: corresponding segments of the waveforms are superimposed to obtain an average waveform value for each run having an increased signal to noise ratio over the individual waveform segments. Also, preferably, a software-created variable frequency digital filter is used to filter signal responses at the cross-correlation frequency.

8. 发明授权

US08692998B2 Apparatus and method for light emission detection for in-depth imaging of turbid media 有权
标题翻译：用于浑浊介质深度成像的发光检测装置和方法
公开(公告)号：US08692998B2
公开(公告)日：2014-04-08
申请号：US13444696
申请日：2012-04-11
申请人： Enrico Gratton , Alexander Dvornikov , Viera Crosignani
发明人： Enrico Gratton , Alexander Dvornikov , Viera Crosignani
IPC分类号： G01N21/00
CPC分类号： G01N21/6456 , G01N21/4795
摘要： An apparatus and method for in-depth fluorescence imaging using two-photon fluorescence imaging in turbid media. The apparatus includes a detector which can significantly enhance the use of a detection method that allows to efficiently collect scattered fluorescence photons from a wide area of the turbid sample. By using this detector it is possible to perform imaging of turbid samples, simulating brain tissue at depths up to 3 mm, where the two-photon induced fluorescence signal is too weak to be detected by previous means used in conventional two-photon microscopy. The detector separates the excitation and detection optics which allows a more efficient collection of fluorescence and enhancing the possible imaging depth.
摘要翻译：用于在混浊介质中使用双光子荧光成像进行深度荧光成像的装置和方法。该装置包括能够显着增强检测方法的使用的检测器，该检测方法允许有效地从浑浊样品的广泛区域收集散射的荧光光子。通过使用该检测器，可以进行浑浊样品的成像，模拟高达3mm深度的脑组织，其中双光子诱导的荧光信号太弱，无法通过传统双光子显微镜中使用的先前手段检测。检测器分离激发和检测光学器件，其允许更有效地收集荧光并增强可能的成像深度。

9. 发明授权

US07528384B2 Methods and devices for characterizing particles in clear and turbid media 有权
标题翻译：在透明和混浊介质中表征颗粒的方法和装置
公开(公告)号：US07528384B2
公开(公告)日：2009-05-05
申请号：US11342273
申请日：2006-01-27
申请人： Enrico Gratton , Guido Motolese , Abdel Tahari
发明人： Enrico Gratton , Guido Motolese , Abdel Tahari
IPC分类号： G01N21/64
CPC分类号： G01N15/1463 , G01N21/6408 , G01N21/6428 , G01N21/6458 , G01N2015/0088 , G01N2015/1486 , G01N2015/1497 , G01N2021/6482
摘要： The invention provides methods and devices for detecting, identifying, classifying and characterizing particles in a fluid sample. Optical analyzers are provided having a rotating and/or translating sample container for measuring the concentrations of fluorescent particles present in very low concentrations and for characterizing fluorescent particles on the basis of size, shape, diffusion constant and/or composition. Scanning optical analyzers are provided using pattern recognitions data analysis techniques and multichannel detection.
摘要翻译：本发明提供用于检测，识别，分类和表征流体样品中的颗粒的方法和装置。提供具有旋转和/或平移样品容器的光学分析器，用于测量以非常低的浓度存在的荧光颗粒的浓度，并且基于尺寸，形状，扩散常数和/或组成来表征荧光颗粒。使用模式识别数据分析技术和多通道检测来提供扫描光学分析仪。

10. 发明申请

US20080009748A1 METHOD AND APPARATUS FOR THE DETERMINATION OF INTRINSIC SPECTROSCOPIC TUMOR MARKERS BY BROADBAND-FREQUENCY DOMAIN TECHNOLOGY 审中-公开
标题翻译：用于通过宽带频域技术确定内在光谱肿瘤标记物的方法和装置
公开(公告)号：US20080009748A1
公开(公告)日：2008-01-10
申请号：US11749704
申请日：2007-05-16
申请人： Enrico Gratton , Shwayta Kukreti , Albert Cerussi , Bruce Tromburg
发明人： Enrico Gratton , Shwayta Kukreti , Albert Cerussi , Bruce Tromburg
IPC分类号： A61B6/00
CPC分类号： A61B5/0059 , A61B5/06 , G01N21/359
摘要： The illustrated embodiment is an improvement in a method of optically analyzing tissue in vivo in an individual to obtain a unique spectrum for the tissue of the individual, the improvement including the steps of optically measuring the tissue of the individual to obtain a spectrum of an optical parameter, and identifying a spectral signature specific to a metabolic or physiologic state in the tissue of the individual with a unique spectrum for the tissue by considering only the spectral differences between a first metabolic or physiologic state of the tissue of the individual and one or more other metabolic or physiologic states of the tissue of the individual such that identification of the spectral signature is self-referencing with respect to intra-individual metabolic or physiologic variations. The method also includes separating benign and malignant lesions only using the shape or a characteristic of the spectrum.
摘要翻译：所示实施例是在个体中在体内光学分析组织以获得个体组织的独特光谱的方法的改进，改进包括光学测量个体的组织以获得光学的光谱的步骤参数，并且通过仅考虑个体组织的第一代谢或生理状态与一个或多个组织之间的光谱差异，通过组织的唯一光谱来识别特定于个体组织中代谢或生理状态的光谱特征个体组织的其他代谢或生理状态，使得光谱特征的鉴定在个体内代谢或生理变化方面是自参照的。该方法还包括仅使用光谱的形状或特征分离良性和恶性病变。

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