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1. 发明授权

US08715572B2 Method and apparatus for detection, analysis, and collection of rare cellular events 有权
标题翻译：用于检测，分析和收集罕见细胞事件的方法和装置
公开(公告)号：US08715572B2
公开(公告)日：2014-05-06
申请号：US13473495
申请日：2012-05-16
申请人： Jiong Wu , Mahesh Junnarkar , Giacomo Vacca
发明人： Jiong Wu , Mahesh Junnarkar , Giacomo Vacca
IPC分类号： G01N21/64 , G01N15/06 , G06K9/00 , G01N15/02 , G01N21/49 , G01N21/85 , G01J3/30 , G01N21/00
CPC分类号： G01N15/1459 , G01N15/1434 , G01N21/05 , G01N21/53 , G01N21/6428 , G01N21/645 , G01N2021/0346 , G01N2021/6482 , G01N2021/6493
摘要： Systems and methods for the detection, analysis, and collection of rare cellular events, wherein rare cellular events are defined by events comprising less than 5% of a total number of cells in a sample. The systems and methods generally include: (1) a flow cell dimensioned so as to permit a flow of a sample through the flow cell at a flow rate greater than 300,000 cells per second; (2) a laser positioned to emit a laser beam directed to the flow cell; (3) one or more deflector components disposed between the laser and the flow cell, wherein the deflector component is configured to affect a position of the laser beam relative to the sample flow; (4) one or more fluorescence emission detectors; and (5) one or more processor configured to detect rare cellular events based on fluorescence emission from cell-binding surface markers introduced into the sample prior to the sample being flowed through the flow cell.
摘要翻译：用于检测，分析和收集罕见细胞事件的系统和方法，其中罕见的细胞事件由包含小于样品中总细胞数量的5％的事件定义。系统和方法通常包括：（1）流动池，其尺寸设计成允许样品以大于每秒300,000个细胞的流速流过流动池; （2）定位成发射指向流动池的激光束的激光器; （3）设置在所述激光器和所述流动池之间的一个或多个偏转器部件，其中所述偏转器部件被配置为影响所述激光束相对于所述样品流的位置; （4）一个或多个荧光发射检测器; 以及（5）一个或多个处理器，被配置为基于在样品流过所述流动池之前引入到样品中的来自细胞结合表面标记物的荧光发射来检测罕见的细胞事件。

2. 发明申请

US20120295339A1 Method and Apparatus for Detection, Analysis, and Collection of Rare Cellular Events 有权
标题翻译：用于检测，分析和收集稀有细胞事件的方法和装置
公开(公告)号：US20120295339A1
公开(公告)日：2012-11-22
申请号：US13473495
申请日：2012-05-16
申请人： Jiong Wu , Mahesh Junnarkar , Giacomo Vacca
发明人： Jiong Wu , Mahesh Junnarkar , Giacomo Vacca
IPC分类号： C12M1/42 , C12M1/36
CPC分类号： G01N15/1459 , G01N15/1434 , G01N21/05 , G01N21/53 , G01N21/6428 , G01N21/645 , G01N2021/0346 , G01N2021/6482 , G01N2021/6493
摘要： Systems and methods for the detection, analysis, and collection of rare cellular events, wherein rare cellular events are defined by events comprising less than 5% of a total number of cells in a sample. The systems and methods generally include: (1) a flow cell dimensioned so as to permit a flow of a sample through the flow cell at a flow rate greater than 300,000 cells per second; (2) a laser positioned to emit a laser beam directed to the flow cell; (3) one or more deflector components disposed between the laser and the flow cell, wherein the deflector component is configured to affect a position of the laser beam relative to the sample flow; (4) one or more fluorescence emission detectors; and (5) one or more processor configured to detect rare cellular events based on fluorescence emission from cell-binding surface markers introduced into the sample prior to the sample being flowed through the flow cell.
摘要翻译：用于检测，分析和收集罕见细胞事件的系统和方法，其中罕见的细胞事件由包含小于样品中总细胞数量的5％的事件定义。系统和方法通常包括：（1）流动池，其尺寸设计成允许样品以大于每秒300,000个细胞的流速流过流动池; （2）定位成发射指向流动池的激光束的激光器; （3）设置在所述激光器和所述流动池之间的一个或多个偏转器部件，其中所述偏转器部件被配置为影响所述激光束相对于所述样品流的位置; （4）一个或多个荧光发射检测器; 以及（5）一个或多个处理器，被配置为基于在样品流过所述流动池之前引入到样品中的来自细胞结合表面标记的荧光发射来检测罕见的细胞事件。

3. 发明授权

US09091624B2 White blood cell analysis system and method 有权
标题翻译：白细胞分析系统及方法
公开(公告)号：US09091624B2
公开(公告)日：2015-07-28
申请号：US13456729
申请日：2012-04-26
申请人： Jiong Wu , Giacomo Vacca
发明人： Jiong Wu , Giacomo Vacca
IPC分类号： G01N15/14 , G01N15/10
CPC分类号： G01N15/1434 , G01N15/1459 , G01N21/6428 , G01N33/49 , G01N2015/008 , G01N2015/1006 , G01N2015/1477 , G01N2015/1488 , G01N2021/6439
摘要： Systems and methods for analyzing blood samples, and more specifically for performing a white blood cell (WBC) differential analysis. The systems and methods screen WBCs by means of fluorescence staining and a fluorescence triggering strategy. As such, interference from unlysed red blood cells (RBCs) and fragments of lysed RBCs is substantially eliminated. The systems and methods also enable development of relatively milder WBC reagent(s), suitable for assays of samples containing fragile WBCs. In one embodiment, the systems and methods include: (a) staining a blood sample with an exclusive cell membrane permeable fluorescent dye, which corresponds in emission spectrum to an excitation source of a hematology instrument; (b) using a fluorescence trigger to screen the blood sample for WBCs; and (c) using measurements of (1) axial light loss, (2) intermediate angle scatter, (3) 90° polarized side scatter, (4) 90° depolarized side scatter, and (5) fluorescence emission to perform a differentiation analysis.
摘要翻译：用于分析血液样品的系统和方法，更具体来说用于进行白细胞（WBC）差异分析。系统和方法通过荧光染色和荧光触发策略筛选WBC。因此，基本上消除了来自未分解的红细胞（RBC）和裂解的RBC的片段的干扰。这些系统和方法还可以开发出相对温和的WBC试剂，适用于含有脆弱WBC的样品的测定。在一个实施方案中，系统和方法包括：（a）用发射光谱对应于血液学仪器的激发源的专用细胞膜可渗透荧光染料染色血液样品; （b）使用荧光触发来筛选白细胞的血样; 和（c）使用（1）轴向光损失，（2）中角散射，（3）90°偏振侧散射，（4）90°去极化侧散射的测量，和（5）荧光发射来进行微分分析。

4. 发明申请

US20120282600A1 Basophil Analysis System and Method 有权
标题翻译：嗜碱粒细胞分析系统和方法
公开(公告)号：US20120282600A1
公开(公告)日：2012-11-08
申请号：US13456744
申请日：2012-04-26
申请人： Jiong Wu , Giacomo Vacca , Michael R. Buhl
发明人： Jiong Wu , Giacomo Vacca , Michael R. Buhl
IPC分类号： C12M1/42 , G01N21/64
CPC分类号： G01N15/1434 , G01N15/1459 , G01N21/6428 , G01N33/49 , G01N2015/0069 , G01N2015/008 , G01N2015/1006 , G01N2015/1402 , G01N2015/1477 , G01N2015/1486 , G01N2015/1488 , G01N2021/6439
摘要： Provided herein are systems and methods for analyzing blood samples, and more specifically for performing a basophil analysis. In one embodiment, the systems and methods include: (a) staining a blood sample with an exclusive cell membrane permeable fluorescent dye; and then (b) using measurements of light scatter and fluorescence emission to distinguish basophils from other WBC sub-populations. In one embodiment, the systems and methods include performing a basophil cluster analysis of the blood sample, based on the combination of light scatter and fluorescence measurements.
摘要翻译：本文提供了用于分析血液样品的系统和方法，并且更具体地用于进行嗜碱性粒细胞分析。在一个实施方案中，系统和方法包括：（a）用专用细胞膜可渗透荧光染料染色血液样品; 然后（b）使用光散射和荧光发射的测量来区分嗜碱性粒细胞与其他WBC亚种群。在一个实施方案中，系统和方法包括基于光散射和荧光测量的组合进行血液样品的嗜碱性粒细胞聚类分析。

5. 发明授权

US09091625B2 Basophil analysis system and method 有权
标题翻译：嗜碱性粒细胞分析系统及方法
公开(公告)号：US09091625B2
公开(公告)日：2015-07-28
申请号：US13456744
申请日：2012-04-26
申请人： Jiong Wu , Giacomo Vacca , Michael R. Buhl
发明人： Jiong Wu , Giacomo Vacca , Michael R. Buhl
IPC分类号： G01N15/14 , G01N15/10
CPC分类号： G01N15/1434 , G01N15/1459 , G01N21/6428 , G01N33/49 , G01N2015/0069 , G01N2015/008 , G01N2015/1006 , G01N2015/1402 , G01N2015/1477 , G01N2015/1486 , G01N2015/1488 , G01N2021/6439
摘要： Provided herein are systems and methods for analyzing blood samples, and more specifically for performing a basophil analysis. In one embodiment, the systems and methods include: (a) staining a blood sample with an exclusive cell membrane permeable fluorescent dye; and then (b) using measurements of light scatter and fluorescence emission to distinguish basophils from other WBC sub-populations. In one embodiment, the systems and methods include performing a basophil cluster analysis of the blood sample, based on the combination of light scatter and fluorescence measurements.
摘要翻译：本文提供了用于分析血液样品的系统和方法，并且更具体地用于进行嗜碱性粒细胞分析。在一个实施方案中，系统和方法包括：（a）用专用细胞膜可渗透荧光染料染色血液样品; 然后（b）使用光散射和荧光发射的测量来区分嗜碱性粒细胞与其他WBC亚种群。在一个实施方案中，系统和方法包括基于光散射和荧光测量的组合进行血液样品的嗜碱性粒细胞聚类分析。

6. 发明授权

US08906309B2 Method for discriminating red blood cells from white blood cells by using forward scattering from a laser in an automated hematology analyzer 有权
标题翻译：在自动血液分析仪中使用来自激光的前向散射来鉴别红细胞与白细胞的方法
公开(公告)号：US08906309B2
公开(公告)日：2014-12-09
申请号：US12767611
申请日：2010-04-26
申请人： Martin Krockenberger , Jiong Wu , Bodo Roemer , Giacomo Vacca
发明人： Martin Krockenberger , Jiong Wu , Bodo Roemer , Giacomo Vacca
IPC分类号： G01N33/00 , G01N31/00 , G01N15/00 , G01N15/14 , G01N33/49
CPC分类号： G01N15/1404 , G01N15/00 , G01N15/1429 , G01N15/1459 , G01N21/53 , G01N33/49 , G01N33/4915 , G01N33/492 , G01N2015/0073 , G01N2015/0076 , G01N2015/008 , G01N2015/0084 , G01N2015/1006 , G01N2015/1486 , G01N2800/22 , Y10T436/101666
摘要： A method for identifying, analyzing, and quantifying the cellular components of whole blood by use of an automated hematology analyzer and the detection of the light scattered, absorbed, and fluorescently emitted by each cell. More particularly, the aforementioned method involves identifying, analyzing, and quantifying the cellular components of whole blood by use of a light source having a wavelength ranging from about 400 nm to about 450 nm and multiple in-flow optical measurements and staining without the need for lysing red blood cells.
摘要翻译：一种通过使用自动血液分析仪来鉴定，分析和定量全血细胞成分的方法，以及每个细胞散射，吸收和荧光发射的光的检测。更具体地，上述方法包括通过使用波长范围为约400nm至约450nm的光源和多次流入光学测量和染色来识别，分析和定量全血的细胞组分，并且不需要裂解红细胞。

7. 发明申请

US20120282599A1 Nucleated Red Blood Cell Analysis System and Method 有权
标题翻译：成核红细胞分析系统及方法
公开(公告)号：US20120282599A1
公开(公告)日：2012-11-08
申请号：US13456738
申请日：2012-04-26
申请人： Jiong Wu , Marilou Coleman , Michael R. Buhl , Giacomo Vacca , Emily H. Lin
发明人： Jiong Wu , Marilou Coleman , Michael R. Buhl , Giacomo Vacca , Emily H. Lin
IPC分类号： C12M1/42 , G01N21/64
CPC分类号： G01N15/1434 , G01N15/1459 , G01N15/147 , G01N21/6428 , G01N33/49 , G01N33/80 , G01N2015/0069 , G01N2015/008 , G01N2015/1006 , G01N2015/1402 , G01N2015/1477 , G01N2015/1488 , G01N2021/6439
摘要： Systems and methods for analyzing blood samples, and more specifically for performing a nucleated red blood cell (nRBC) analysis. The systems and methods screen a blood sample by means of fluorescence staining and a fluorescence triggering strategy, to identify nuclei-containing particles within the blood sample. As such, interference from unlysed red blood cells (RBCs) and fragments of lysed RBCs is substantially eliminated. The systems and methods also enable development of relatively milder reagent(s), suitable for assays of samples containing fragile white blood cells (WBCs). In one embodiment, the systems and methods include: (a) staining a blood sample with an exclusive cell membrane permeable fluorescent dye; (b) using a fluorescence trigger to screen the blood sample for nuclei-containing particles; and (c) using measurements of light scatter and fluorescence emission to distinguish nRBCs from WBCs.
摘要翻译：用于分析血液样品的系统和方法，更具体地用于进行有核红细胞（nRBC）分析。系统和方法通过荧光染色和荧光触发策略筛选血液样品，以鉴定血液样品中含有核的颗粒。因此，基本上消除了来自未分解的红细胞（RBC）和裂解的RBC的片段的干扰。这些系统和方法还能够开发适合于含有易碎白细胞（WBC）的样品的测定的较温和的试剂。在一个实施方案中，系统和方法包括：（a）用专用细胞膜可渗透荧光染料染色血液样品; （b）使用荧光触发器来筛选含有核的颗粒的血液样品; 和（c）使用光散射和荧光发射的测量来区分来自WBC的nRBC。

8. 发明授权

US09097704B2 Method for hematology analysis 有权
标题翻译：血液学分析方法
公开(公告)号：US09097704B2
公开(公告)日：2015-08-04
申请号：US13093502
申请日：2011-04-25
申请人： Jiong Wu , Giacomo Vacca
发明人： Jiong Wu , Giacomo Vacca
IPC分类号： G01N15/14 , G01N15/10 , G01N33/50 , G01N33/58 , G01N15/00
CPC分类号： G01N15/1434 , G01N15/147 , G01N21/6428 , G01N33/5047 , G01N33/5094 , G01N33/582 , G01N35/1002 , G01N2015/0065 , G01N2015/0069 , G01N2015/1006 , G01N2015/1486 , G01N2015/149 , G01N2021/6439
摘要： A method whereby one or more fluorescent dyes are used to bind and stain nucleic acids in certain blood cells, such as, for example, white blood cells, nucleated red blood cells, and reticulocytes, and to induce fluorescent emissions upon excitation of photons from a given source of light, such as, for example, a laser, at an appropriate wavelength. More particularly, this invention provides a method whereby a fluorescent trigger is used in a data collection step for collecting events that emit strong fluorescence, in order to separate white blood cells and nucleated red blood cells from red blood cells and platelets without the need for using a lysing agent.
摘要翻译：一种或多种荧光染料用于结合和染色某些血细胞（例如白细胞，有核红细胞和网织红细胞）中的核酸的方法，以及在从给定光源，例如激光，以适当的波长。更具体地，本发明提供了一种方法，其中在用于收集发射强荧光的事件的数据收集步骤中使用荧光触发器，以将白细胞和有核红细胞与红细胞和血小板分离而不需要使用裂解剂。

9. 发明申请

US20120282598A1 White Blood Cell Analysis System and Method 有权
标题翻译：白细胞分析系统及方法
公开(公告)号：US20120282598A1
公开(公告)日：2012-11-08
申请号：US13456729
申请日：2012-04-26
申请人： Jiong Wu , Giacomo Vacca
发明人： Jiong Wu , Giacomo Vacca
IPC分类号： C12M1/34 , G01N21/64
CPC分类号： G01N15/1434 , G01N15/1459 , G01N21/6428 , G01N33/49 , G01N2015/008 , G01N2015/1006 , G01N2015/1477 , G01N2015/1488 , G01N2021/6439
摘要： Systems and methods for analyzing blood samples, and more specifically for performing a white blood cell (WBC) differential analysis. The systems and methods screen WBCs by means of fluorescence staining and a fluorescence triggering strategy. As such, interference from unlysed red blood cells (RBCs) and fragments of lysed RBCs is substantially eliminated. The systems and methods also enable development of relatively milder WBC reagent(s), suitable for assays of samples containing fragile WBCs. In one embodiment, the systems and methods include: (a) staining a blood sample with an exclusive cell membrane permeable fluorescent dye, which corresponds in emission spectrum to an excitation source of a hematology instrument; (b) using a fluorescence trigger to screen the blood sample for WB Cs; and (c) using measurements of (1) axial light loss, (2) intermediate angle scatter, (3) 90° polarized side scatter, (4) 90° depolarized side scatter, and (5) fluorescence emission to perform a differentiation analysis.
摘要翻译：用于分析血液样品的系统和方法，更具体来说用于进行白细胞（WBC）差异分析。系统和方法通过荧光染色和荧光触发策略筛选WBC。因此，基本上消除了来自未分解的红细胞（RBC）和裂解的RBC的片段的干扰。这些系统和方法还可以开发出相对温和的WBC试剂，适用于含有脆弱WBC的样品的测定。在一个实施方案中，系统和方法包括：（a）用发射光谱对应于血液学仪器的激发源的专用细胞膜可渗透荧光染料染色血液样品; （b）使用荧光触发器筛选WB Cs的血样; 和（c）使用（1）轴向光损失，（2）中角散射，（3）90°偏振侧散射，（4）90°去极化侧散射的测量，和（5）荧光发射来进行微分分析。

10. 发明申请

US20110275064A1 METHOD FOR HEMATOLOGY ANALYSIS 有权
标题翻译：血液学分析方法
公开(公告)号：US20110275064A1
公开(公告)日：2011-11-10
申请号：US13093502
申请日：2011-04-25
申请人： Jiong Wu , Giacomo Vacca
发明人： Jiong Wu , Giacomo Vacca
IPC分类号： C12Q1/68
CPC分类号： G01N15/1434 , G01N15/147 , G01N21/6428 , G01N33/5047 , G01N33/5094 , G01N33/582 , G01N35/1002 , G01N2015/0065 , G01N2015/0069 , G01N2015/1006 , G01N2015/1486 , G01N2015/149 , G01N2021/6439
摘要： A method whereby one or more fluorescent dyes are used to bind and stain nucleic acids in certain blood cells, such as, for example, white blood cells, nucleated red blood cells, and reticulocytes, and to induce fluorescent emissions upon excitation of photons from a given source of light, such as, for example, a laser, at an appropriate wavelength. More particularly, this invention provides a method whereby a fluorescent trigger is used in a data collection step for collecting events that emit strong fluorescence, in order to separate white blood cells and nucleated red blood cells from red blood cells and platelets without the need for using a lysing agent.
摘要翻译：一种或多种荧光染料用于结合和染色某些血细胞（例如白细胞，有核红细胞和网织红细胞）中的核酸的方法，以及在从给定光源，例如激光，以适当的波长。更具体地，本发明提供了一种方法，其中在用于收集发射强荧光的事件的数据收集步骤中使用荧光触发器，以将白细胞和有核红细胞与红细胞和血小板分离而不需要使用裂解剂。

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