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1.发明授权
公开(公告)号：US6049380A
公开(公告)日：2000-04-11
申请号：US169025
申请日：1998-10-09
CPC分类号： G01N21/6428 , G01N21/6408
摘要： Single fluorescent molecules in a flowing sample stream are distinguished and identified using only a single laser excitation wavelength. A sample stream is formed containing a dilute mixture of single molecule fluorophores, wherein each one of the fluorophores is serially ordered in the sample stream. The sample stream is illuminated with s single excitation wavelength laser effective to excite each fluorophore one at a time. Fluorescence emission photons from each said fluorophore are detected. A burst size is determined for each fluorophore to identify each fluorophore. A pulsed laser may be used, where burst size and an intra-burst fluorescence decay rate for each fluorophore are determined simultaneously from the detected fluorescence emission photons. The burst size and the decay rate are correlated to identify each fluorophore.
摘要翻译： 流动样品流中的单个荧光分子仅使用单个激光激发波长来识别和识别。 形成包含单分子荧光团的稀混合物的样品流，其中每个荧光团在样品流中连续排列。 用单个激发波长激光器照射样品流，有效地一次激发每个荧光团。 检测来自每个所述荧光团的荧光发射光子。 确定每个荧光团的突发尺寸以识别每个荧光团。 可以使用脉冲激光，其中从检测到的荧光发射光子同时测定每个荧光团的突发尺寸和爆发内荧光衰减速率。 脉冲串大小和衰减速率相关，以识别每个荧光团。
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公开(公告)号：US06309886B1
公开(公告)日：2001-10-30
申请号：US09449280
申请日：1999-11-24
申请人： W. Patrick Ambrose , W. Kevin Grace , Peter M. Goodwin , James H. Jett , Alan Van Orden , Richard A. Keller
发明人： W. Patrick Ambrose , W. Kevin Grace , Peter M. Goodwin , James H. Jett , Alan Van Orden , Richard A. Keller
IPC分类号： G01N2164
CPC分类号： G01N21/6456 , G01N15/1434 , G01N15/1459 , G01N21/6458 , G01N2015/1006
摘要： Apparatus and method enable imaging multiple fluorescent sample particles in a single flow channel. A flow channel defines a flow direction for samples in a flow stream and has a viewing plane perpendicular to the flow direction. A laser beam is formed as a ribbon having a width effective to cover the viewing plane. Imaging optics are arranged to view the viewing plane to form an image of the fluorescent sample particles in the flow stream, and a camera records the image formed by the imaging optics.
摘要翻译： 装置和方法能够在单个流动通道中成像多个荧光样品颗粒。 流动通道限定流动流中的样品的流动方向，并且具有垂直于流动方向的观察平面。 激光束形成为具有有效覆盖观察平面的宽度的带状物。 成像光学器件被布置成观看观察平面以形成流动流中的荧光样品颗粒的图像，并且相机记录由成像光学元件形成的图像。
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公开(公告)号：US5558998A
公开(公告)日：1996-09-24
申请号：US486400
申请日：1995-06-05
IPC分类号： C12Q1/68 , G01N15/14 , G01N33/483 , G01N35/08
CPC分类号： C12Q1/6827 , G01N15/14 , Y10T436/117497
摘要： A method is provided for sizing DNA fragments using high speed detection systems, such as flow cytometry to determine unique characteristics of DNA pieces from a sample. In one characterization the DNA piece is fragmented at preselected sites to produce a plurality of DNA fragments. The DNA piece or the resulting DNA fragments are treated with a dye effective to stain stoichiometrically the DNA piece or the DNA fragments. The fluorescence from the dye in the stained fragments is then examined to generate an output functionally related to the number of nucleotides in each one of the DNA fragments. In one embodiment, the intensity of the fluorescence emissions from each fragment is linearly related to the fragment length. The distribution of DNA fragment sizes forms a characterization of the DNA piece for use in forensic and research applications.
摘要翻译： 提供了使用高速检测系统（例如流式细胞术）来定尺寸DNA片段的方法，以确定样品中DNA片段的独特特征。 在一个表征中，DNA片段在预选位点被片段化以产生多个DNA片段。 用有效染色DNA片段或DNA片段化学计量的染料处理DNA片段或得到的DNA片段。 然后检查染色片段中来自染料的荧光，以产生功能上与每个DNA片段中的核苷酸数相关的输出。 在一个实施方案中，来自每个片段的荧光发射强度与片段长度线性相关。 DNA片段大小的分布形成了用于法医学和研究应用的DNA片段的表征。
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4.发明授权Method for rapid base sequencing in DNA and RNA with two base labeling 失效用于在两个碱基标记的DNA和RNA中进行快速碱基测序的方法
公开(公告)号：US5405747A
公开(公告)日：1995-04-11
申请号：US208506
申请日：1994-03-07
申请人： James H. Jett , Richard A. Keller , John C. Martin , Richard G. Posner , Babetta L. Marrone , Mark L. Hammond , Daniel J. Simpson
发明人： James H. Jett , Richard A. Keller , John C. Martin , Richard G. Posner , Babetta L. Marrone , Mark L. Hammond , Daniel J. Simpson
CPC分类号： C12Q1/6816 , C12Q1/6869 , Y10T436/143333
摘要： Method for rapid-base sequencing in DNA and RNA with two-base labeling and employing fluorescent detection of single molecules at two wavelengths. Bases modified to accept fluorescent labels are used to replicate a single DNA or RNA strand to be sequenced. The bases are then sequentially cleaved from the replicated strand, excited with a chosen spectrum of electromagnetic radiation, and the fluorescence from individual, tagged bases detected in the order of cleavage from the strand.
摘要翻译： 在DNA和RNA中进行快速碱基测序的方法，采用双碱基标记，并采用双波长单分子荧光检测。 用于接受荧光标记的碱基用于复制要测序的单个DNA或RNA链。 然后从复制的链中顺序地切割碱基，用所选择的电磁辐射谱激发，并且以从链断裂的顺序检测来自单个的标记碱基的荧光。
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公开(公告)号：US4962037A
公开(公告)日：1990-10-09
申请号：US105375
申请日：1987-10-07
申请人： James H. Jett , Richard A. Keller , John C. Martin , Robert K. Moyzis , Robert L. Ratliff , E. Brooks Shera , Carleton C. Stewart
发明人： James H. Jett , Richard A. Keller , John C. Martin , Robert K. Moyzis , Robert L. Ratliff , E. Brooks Shera , Carleton C. Stewart
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6869 , C12Q1/6813 , C12Q1/6823 , Y10S436/80 , Y10T436/143333 , Y10T436/25125
摘要： A method is provided for the rapid base sequencing of DNA or RNA fragments wherein a single fragment of DNA or RNA is provided with identifiable bases and suspended in a moving flow stream. An exonuclease sequentially cleaves individual bases from the end of the suspended fragment. The moving flow stream maintains the cleaved bases in an orderly train for subsequent detection and identification. In a particular embodiment, individual bases forming the DNA or RNA fragments are individually tagged with a characteristic fluorescent dye. The train of bases is then excited to fluorescence with an output spectrum characteristic of the individual bases. Accordingly, the base sequence of the original DNA or RNA fragment can be reconstructed.
摘要翻译： 提供了用于DNA或RNA片段的快速碱基测序的方法，其中DNA或RNA的单个片段具有可鉴定的碱基并悬浮在移动的流动中。 外切核酸酶从悬浮片段的末端依次切割各个碱基。 移动流动流将切割的基部保持在有序的列中，用于随后的检测和识别。 在一个具体实施方案中，形成DNA或RNA片段的单个碱基用特征荧光染料单独标记。 然后将基因列激发成具有各基因的输出光谱特征的荧光。 因此，可以重构原始DNA或RNA片段的碱基序列。
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公开(公告)号：US5879625A
公开(公告)日：1999-03-09
申请号：US951955
申请日：1997-10-17
CPC分类号： G01N15/1459 , C12N15/1006 , C12Q1/68 , G01N2015/0038 , G01N2015/149 , Y10T436/143333
摘要： Optical selection and collection of DNA fragments. The present invention includes the optical selection and collection of large (>.mu.g) quantities of clonable, chromosome-specific DNA from a sample of chromosomes. Chromosome selection is based on selective, irreversible photoinactivation of unwanted chromosomal DNA. Although more general procedures may be envisioned, the invention is demonstrated by processing chromosomes in a conventional flow cytometry apparatus, but where no droplets are generated. All chromosomes in the sample are first stained with at least one fluorescent analytic dye and bonded to a photochemically active species which can render chromosomal DNA unclonable if activated. After passing through analyzing light beam(s), unwanted chromosomes are irradiated using light which is absorbed by the photochemically active species, thereby causing photoinactivation. As desired chromosomes pass this photoinactivation point, the inactivating light source is deflected by an optical modulator; hence, desired chromosomes are not photoinactivated and remain clonable. The selection and photoinactivation processes take place on a microsecond timescale. By eliminating droplet formation, chromosome selection rates 50 times greater than those possible with conventional chromosome sorters may be obtained. Thus, usable quantities of clonable DNA from any source thereof may be collected.
摘要翻译： DNA片段的光学选择和收集。 本发明包括从染色体样品中大量（>μg）量的可克隆，染色体特异性DNA的光学选择和收集。 染色体选择基于不需要的染色体DNA的选择性不可逆光激活。 尽管可以设想更一般的程序，但是通过在常规流式细胞术装置中处理染色体但是不产生液滴来证明本发明。 样品中的所有染色体首先用至少一种荧光分析染料染色，并与光化学活性物质结合，如果激活可使染色体DNA不可克隆。 在通过分析光束之后，使用被光化学活性物质吸收的光照射不想要的染色体，从而引起光激活。 根据需要，染色体通过该光激活点，灭活光源被光学调制器偏转; 因此，期望的染色体不是光激活的并且保持克隆。 选择和光激活过程在微秒的时间尺度上进行。 通过消除液滴形成，可以获得比常规染色体分选机可能的染色体选择率高50倍的染色体选择率。 因此，可以收集来自其任何来源的可克隆DNA的可用量。
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公开(公告)号：US5085673A
公开(公告)日：1992-02-04
申请号：US671330
申请日：1991-03-19
IPC分类号： B03C3/014
CPC分类号： B03C3/014
摘要： Method and apparatus for removing material from a gas. A mist created by a piezoelectric ultrasonic transducer is contacted with the gas and both gas and mist are passed through baffled separators. Liquid effluent from the separators contains solid material removed from the gas and gaseous material which reacted with the liquid or was absorbed by the liquid. The invention is useful for collecting a sample of material in a gas, such as a vapor in the atmosphere, and in cleaning a gas. A relatively concentrated solution of a material present in a gas in a very small concentration can be obtained.
摘要翻译： 从气体中去除材料的方法和装置。 由压电超声换能器产生的雾气与气体接触，气体和雾气均通过挡板分离器。 来自分离器的液体流出物包含从与液体反应或被液体吸收的气体和气态物质中去除的固体材料。 本发明可用于收集诸如气氛中的气体的气体中的材料样品和清洁气体。 可以获得以非常小的浓度存在于气体中的材料的相对浓缩的溶液。
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公开(公告)号：US5707808A
公开(公告)日：1998-01-13
申请号：US632743
申请日：1996-04-15
CPC分类号： G01N15/1459 , C12N15/1006 , C12Q1/68 , G01N2015/0038 , G01N2015/149 , Y10T436/143333
摘要： Optical selection and collection of DNA fragments. The present invention includes the optical selection and collection of large (>.mu.g) quantities of clonable, chromosome-specific DNA from a sample of chromosomes. Chromosome selection is based on selective, irreversible photoinactivation of unwanted chromosomal DNA. Although more general procedures may be envisioned, the invention is demonstrated by processing chromosomes in a conventional flow cytometry apparatus, but where no droplets are generated. All chromosomes in the sample are first stained with at least one fluorescent analytic dye and bonded to a photochemically active species which can render chromosomal DNA unclonable if activated. After passing through analyzing light beam(s), unwanted chromosomes are irradiated using light which is absorbed by the photochemically active species, thereby causing photoinactivation. As desired chromosomes pass this photoinactivation point, the inactivating light source is deflected by an optical modulator; hence, desired chromosomes are not photoinactivated and remain clonable. The selection and photoinactivation processes take place on a microsecond timescale. By eliminating droplet formation, chromosome selection rates 50 times greater than those possible with conventional chromosome sorters may be obtained. Thus, usable quantities of clonable DNA from any source thereof may be collected.
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9.发明授权
公开(公告)号：US4573796A
公开(公告)日：1986-03-04
申请号：US568768
申请日：1984-01-06
申请人： John C. Martin , James H. Jett
发明人： John C. Martin , James H. Jett
CPC分类号： G01N21/645 , G01N15/1429 , G01N21/6428 , G01N2015/1438 , G01N2021/6439
摘要： The disclosure is directed to an apparatus for eliminating background interference during fluorescence measurements in a multiple laser flow cytometer. A biological particle stained with fluorescent dyes is excited by a laser. A fluorescence detector detects the fluorescence. The particle scatters light and a gate signal is generated and delayed until the biological particle reaches the next laser. The delayed signal turns on this next laser, which excites a different stained component of the same biological particle.
摘要翻译： 本发明涉及一种用于在多激光流式细胞仪中消除荧光测量期间的背景干扰的装置。 用荧光染料染色的生物颗粒被激光激发。 荧光检测器检测荧光。 颗粒散射光，并产生并延迟栅极信号，直到生物颗粒到达下一个激光。 延迟信号打开下一个激光，激发相同生物粒子的不同染色成分。
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