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1. 发明授权

US06834122B2 Visualization and processing of multidimensional data using prefiltering and sorting criteria 失效
标题翻译：使用预过滤和排序标准可视化和处理多维数据
公开(公告)号：US06834122B2
公开(公告)日：2004-12-21
申请号：US09767595
申请日：2001-01-22
申请人： Mary M. Yang , Edward J. Bylina , William J. Coleman , Michael R. Dilworth , Steven J. Robles , Douglas C. Youvan
发明人： Mary M. Yang , Edward J. Bylina , William J. Coleman , Michael R. Dilworth , Steven J. Robles , Douglas C. Youvan
IPC分类号： G06K968
CPC分类号： G06K9/00127 , G06K9/0014 , G06K9/0063 , G06K9/6253 , G06K2009/00644 , G06K2009/4657 , G06T7/0012 , G06T2200/24 , G06T2207/10016 , G06T2207/10056 , G06T2207/30024 , Y10S128/922 , Y10S707/99937
摘要： Complex multidimensional datasets generated by digital imaging spectroscopy can be organized and analyzed by applying software and computer-based methods comprising sorting algorithms. Combinations of these algorithms to images and graphical data, allow pixels or features to be rapidly and efficiently classified into meaningful groups according to defined criteria. Multiple rounds of pixel or feature selection may be performed based on independent sorting criteria. In one embodiment sorting by spectral criteria (e.g., intensity at a given wavelength) is combined with sorting by temporal criteria (e.g., absorbance at a given time) to identify microcolonies of recombinant organisms harboring mutated genes encoding enzymes having desirable kinetic attributes and substrate specificity. Restriction of the set of pixels analyzed in a subsequent sort based on criteria applied in an earlier sort (“sort and lock” analyses) minimize computational and storage resources. User-defined criteria can also be incorporated into the sorting process by means of a graphical user interface that comprises a visualization tools including a contour plot, a sorting bar and a grouping bar, an image window, and a plot window that allow run-time interactive identification of pixels or features meeting one or more criteria, and display of their associated spectral or kinetic data. These methods are useful for extracting information from imaging data in applications ranging from biology and medicine to remote sensing.
摘要翻译：通过数字成像光谱法生成的复杂多维数据集可以通过应用软件和基于计算机的方法进行组织和分析，包括排序算法。这些算法与图像和图形数据的组合允许像素或特征根据定义的标准快速有效地分类为有意义的组。可以基于独立排序标准来执行多轮像素或特征选择。在一个实施方案中，通过光谱标准（例如，给定波长处的强度）的排序与通过时间标准（例如，给定时间的吸光度）排序相结合，以鉴定携带突变基因的重组体的微菌落，所述突变基因编码具有期望的动力学属性和底物特异性的酶。基于先前排序（“排序和锁定”分析）中应用的标准，在后续排序中分析的像素集合的限制最小化了计算和存储资源。用户定义的标准也可以通过包括可视化工具的图形用户界面结合到分类过程中，该可视化工具包括轮廓图，分类栏和分组条，图像窗口和允许运行时间的绘图窗口满足一个或多个标准的像素或特征的交互式识别，以及它们相关联的光谱或动力学数据的显示。这些方法可用于从生物学和医学到遥感等应用中从成像数据中提取信息。

2. 发明授权

US06661909B2 Calibration of fluorescence resonance energy transfer in microscopy 失效
公开(公告)号：US06661909B2
公开(公告)日：2003-12-09
申请号：US09912117
申请日：2001-07-23
申请人： Dougalas C. Youvan , Christopher M. Silva , Edward J. Bylina , William J. Coleman , Michael R. Dilworth , Mary M. Yang
发明人： Dougalas C. Youvan , Christopher M. Silva , Edward J. Bylina , William J. Coleman , Michael R. Dilworth , Mary M. Yang
IPC分类号： G06K900
CPC分类号： G01N21/6458 , G01N21/278 , G01N21/6428 , G01N2021/6441
摘要： Imaging hardware, software, calibrants, and methods are provided to visualize and quantitate the amount of Fluorescence Resonance Energy Transfer (FRET) occurring between donor and acceptor molecules in epifluorescence microscopy. The MicroFRET system compensates for overlap among donor, acceptor, and FRET spectra using well characterized fluorescent beads as standards in conjunction with radiometrically calibrated image processing techniques. The MicroFRET system also provides precisely machined epifluorescence cubes to maintain proper image registration as the sample is illuminated at the donor and acceptor excitation wavelengths. Algorithms are described that pseudocolor the image to display pixels exhibiting radiometrically-corrected fluorescence emission from the donor (blue), the acceptor (green) and FRET (red). The method is demonstrated on samples exhibiting FRET between genetically engineered derivatives of the Green Fluorescent Protein (GFP) bound to the surface of Ni chelating beads by histidine-tags.

3. 发明授权

US5859700A High resolution imaging microscope (HIRIM) and uses thereof 失效
公开(公告)号：US5859700A
公开(公告)日：1999-01-12
申请号：US562272
申请日：1995-11-22
申请人： Mary M. Yang
发明人： Mary M. Yang
IPC分类号： G01J3/28 , G01N21/25 , G01N21/64 , G02B21/00 , G01J3/00
CPC分类号： G01N21/6458 , G01J3/2823 , G01N21/253 , G02B21/0096 , G01N2021/6423
摘要： The invention pertains to a high resolution imaging microscope, and more specifically, to a digital imaging spectrophotometer (DIS) configured to enable the direct determination of the ground state absorption spectra of microscopic samples from violet to near infrared. The instrument is capable of simultaneously recording hundreds of spectra, and can, using image processing techniques identify features (such as a single microbial cell) on a microscope slide or thin section. The invention additionally relates to the use of such an instrument.

4. 发明授权

US5852498A Optical instrument having a variable optical filter 失效
标题翻译：具有可变滤光器的光学仪器
公开(公告)号：US5852498A
公开(公告)日：1998-12-22
申请号：US833351
申请日：1997-04-04
申请人： Douglas C. Youvan , Mary M. Yang
发明人： Douglas C. Youvan , Mary M. Yang
IPC分类号： G01N21/64 , G01J3/12 , G01J3/28 , G01J3/44 , G02B7/00 , G02B21/00
CPC分类号： G01J3/4406 , G01J3/12 , G01J3/2823 , G02B26/007 , G02B26/008
摘要： An optical instrument for use in collecting light from an object, the optical instrument having a post-objective waist with a variable optical filter positioned approximately at the post-objective waist. The invention also comprises an optical instrument for collecting light from an object, comprising an objective lens positioned to receive light from the object, an exit lens positioned to focus the light at a focal position, and a variable optical filter positioned between the objective lens and the focal position approximately at a waist of the light. The optical instrument may be an epifluorescence microscope for use in observing an image formed from light from an object under illumination by a light source. The light source may be, for example, a filtered light source or a monochromatic light source, preferably in an epifluorescence configuration. The variable optical filter may be a circular variable interference filter or a tilting interference filter. In some embodiments, the orientation of the filter may be varied to block light outside of one band at one time and outside of another band at another time. The variable filter preferably is band pass, blocking substantially all light at wavelengths other than a selected wavelength band.
摘要翻译：一种用于从物体收集光的光学仪器，所述光学仪器具有后物镜腰部，其具有大致位于后物镜腰部的可变滤光器。本发明还包括一种用于收集来自物体的光的光学仪器，包括位于接收来自物体的光的物镜，定位成将光聚焦在焦点位置处的出射透镜和位于物镜和焦点位置大约在光的腰部。光学仪器可以是用于观察由光源照明的物体的光形成的图像的落射荧光显微镜。光源可以是例如滤光光源或单色光源，优选地呈落射荧光结构。可变滤光器可以是圆形可变干涉滤光器或倾斜干涉滤光器。在一些实施例中，可以改变滤波器的取向以在另一时间一次和另一频带之外阻挡一个频带外的光。可变滤波器优选是带通，基本上阻挡除了所选波长带之外的波长的所有光。

5. 发明授权

US06738502B1 Multispectral taxonomic identification 失效
标题翻译：多光谱分类鉴定
公开(公告)号：US06738502B1
公开(公告)日：2004-05-18
申请号：US09585951
申请日：2000-06-02
申请人： William Coleman , Michael A. Tanner , Christopher M. Silva , Edward Bylina , Steven J. Robles , Michael R. Dilworth , Douglas C. Youvan , Mary M. Yang
发明人： William Coleman , Michael A. Tanner , Christopher M. Silva , Edward Bylina , Steven J. Robles , Michael R. Dilworth , Douglas C. Youvan , Mary M. Yang
IPC分类号： G06K900
CPC分类号： G01N21/274 , G01N21/6458 , G01N2201/127
摘要： The present invention provides an instrument and methods for a multispectral optical technique that can simultaneously classify individual biological cells within mixed populations. This invention, known as Multispectral Taxonomic Identification (MTID), shows that microscopy can be combined with a software analysis program to measure and categorize the fluorescence and other spectroscopically identifiable signals from complex populations of cells in situ, without cultivation. The invention thus enables high-throughput screening of cells for taxonomic classification.
摘要翻译：本发明提供了一种用于多光谱光学技术的仪器和方法，其可以在混合群体内同时分类各个生物细胞。称为多光谱分类鉴定（MTID）的本发明表明，显微镜可以与软件分析程序相结合，以便对来自复杂原位细胞群的荧光和其他可光谱鉴定的信号进行原位测量和分类，无需培养。因此，本发明使得能够进行用于分类分类的细胞的高通量筛选。

6. 发明授权

US06456734B1 Calibration of fluorescence resonance energy transfer in microscopy 失效
公开(公告)号：US06456734B1
公开(公告)日：2002-09-24
申请号：US09092316
申请日：1998-06-05
申请人： Douglas C. Youvan , Christopher M. Silva , Edward J. Bylina , William J. Coleman , Michael R. Dilworth , Mary M. Yang
发明人： Douglas C. Youvan , Christopher M. Silva , Edward J. Bylina , William J. Coleman , Michael R. Dilworth , Mary M. Yang
IPC分类号： G06K900
CPC分类号： G01N21/6458 , G01N21/278 , G01N21/6428 , G01N2021/6441
摘要： Imaging hardware, software, calibrants, and methods are provided to visualize and quantitate the amount of Fluorescence Resonance Energy Transfer (FRET) occurring between donor and acceptor molecules in epifluorescence microscopy. The MicroFRET system compensates for overlap among donor, acceptor, and FRET spectra using well characterized fluorescent beads as standards in conjunction with radiometrically calibrated image processing techniques. The MicroFRET system also provides precisely machined epifluorescence cubes to maintain proper image registration as the sample is illuminated at the donor and acceptor excitation wavelengths. Algorithms are described that pseudocolor the image to display pixels exhibiting radiometrically-corrected fluorescence emission from the donor (blue), the acceptor (green) and FRET (red). The method is demonstrated on samples exhibiting FRET between genetically engineered derivatives of the Green Fluorescent Protein (GFP) bound to the surface of Ni chelating beads by histidine-tags.

7. 发明授权

US6160617A High resolution imaging microscope (HIRIM) and uses thereof 失效
标题翻译：高分辨率成像显微镜（HIRIM）及其应用
公开(公告)号：US6160617A
公开(公告)日：2000-12-12
申请号：US229462
申请日：1999-01-12
申请人： Mary M. Yang
发明人： Mary M. Yang
IPC分类号： G01J3/28 , G01N21/25 , G01N21/64 , G02B21/00 , G01J3/00
CPC分类号： G01N21/6458 , G01J3/2823 , G01N21/253 , G01N2021/6423 , G02B21/0096
摘要： The invention pertains to a high resolution imaging microscope, and more specifically, to a digital imaging spectrophotometer (DIS) configured to enable the direct determination of the ground state absorption spectra of microscopic samples from violet to near infrared. The instrument is capable of simultaneously recording hundreds of spectra, and can, using image processing techniques identify features (such as a single microbial cell) on a microscope slide or thin section. The invention additionally relates to the use of such an instrument.
摘要翻译：本发明涉及高分辨率成像显微镜，更具体地说，涉及一种数字成像分光光度计（DIS），其被配置为能够将微观样品的基态吸收光谱直接测定为紫色至近红外。该仪器能够同时记录数百个光谱，并且可以使用图像处理技术在显微镜载玻片或薄片上识别特征（如单个微生物细胞）。本发明还涉及这种仪器的使用。

8. 发明授权

US4514278A Trace water sensor 失效
标题翻译：痕量水传感器
公开(公告)号：US4514278A
公开(公告)日：1985-04-30
申请号：US457992
申请日：1983-01-14
申请人： James B. Stephens , Mary M. Yang , Eric G. Laue
发明人： James B. Stephens , Mary M. Yang , Eric G. Laue
IPC分类号： G01N27/42 , G01N27/49 , G01N27/46
CPC分类号： G01N27/4045 , G01N27/423
摘要： A solid electrolytic type hygrometer is described, which operates with high reliability while providing rapid and sensitive response. A gold foil electrode (16) is wrapped about a hollow glass cylinder (18), a sheet (12) of hygroscopic-electrolytic material is wrapped about the foil, and a wire (14) is wound around the outside of the electrolytic sheet. Moisture passing between wire turns can be absorbed by the electrolytic material (12), and then dissociated by current passed by the electrodes (14, 16) through the electrolytic material. The cylinder has a slit (20) extending along its length, to allow resilient expansion to press the sheet of electrolytic material firmly against the electrodes. The wire turns lie against one another to cause rapid dissociation of moisture throughout the electrolytic material. Additional guard wires (42,44, FIG. 2) lie at opposite ends of the electrolytic sheet, and currents pass through them to avoid moisture buildup at the ends of the main wire coil. The electrical current through the sheet or membrane is proportional to the partial pressure of the water-vapor.
摘要翻译：描述了一种固体电解式湿度计，其以高可靠性运行，同时提供快速和敏感的响应。金箔电极（16）围绕中空玻璃圆筒（18）包裹，吸湿电解材料片（12）缠绕在箔上，并且电线（14）缠绕在电解片的外部。导线之间的水分通过电解材料（12）吸收，然后通过电极（14,16）通过电解质的电流解离。气缸具有沿其长度延伸的狭缝（20），以允许弹性膨胀将电解材料片牢固地压靠在电极上。导线相互靠在一起，导致整个电解材料中的水分迅速离解。附加的防护线（图2中的42,44）位于电解片的相对端，并且电流通过它们以避免在主线圈的端部产生水分积聚。通过片材或膜的电流与水蒸气的分压成比例。

9. 发明授权

US5914245A Solid phase enzyme kinetics screening in microcolonies 失效
标题翻译：微菌落中的固相酶动力学筛选
公开(公告)号：US5914245A
公开(公告)日：1999-06-22
申请号：US98202
申请日：1998-06-16
申请人： Edward J. Bylina , William J. Coleman , Michael R. Dilworth , Christopher M. Silva , Mary M. Yang , Douglas C. Youvan
发明人： Edward J. Bylina , William J. Coleman , Michael R. Dilworth , Christopher M. Silva , Mary M. Yang , Douglas C. Youvan
IPC分类号： C12Q1/04 , C12Q1/34 , C12Q1/44 , C12Q1/00 , C12Q1/37 , C12Q1/54
CPC分类号： C12Q1/04 , C12Q1/34 , Y10S435/808 , Y10S435/968
摘要： A MicroColonyImager instrument and solid phase methods to screen cells expressing mutagenized enzymes for enhanced activity. The MicroColonyImager instrument and methods permit high throughput screening of enzyme libraries by time course analyses of single-pixels, using either absorption, fluorescence or FRET. This high throughput assay can detect small differences in enzyme rates within microcolonies grown at a nearly confluent density on an assay disk. Each microcolony is analyzed simultaneously at single-pixel resolution, requiring less than 100 ml substrate/measurement. By simultaneously assaying different substrates tagged with spectrally distinct chromogenic or fluorogenic reporters, the substrate specificity of an enzyme can be changed.
摘要翻译： MicroColonyImager仪器和固相方法筛选表达诱变酶的细胞以增强活性。 MicroColonyImager仪器和方法允许通过使用吸收，荧光或FRET的单个像素的时程分析来酶库的高通量筛选。这种高通量测定可以检测在测定盘上以几乎汇合的密度生长的微菌落内的酶速度的小差异。每个微绒毛在单像素分辨率下同时进行分析，需要少于100毫升的底物/测量。通过同时测定用光谱不同的显色或荧光报告物标记的不同底物，可以改变酶的底物特异性。

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