公开(公告)号：US07625712B2

公开(公告)日：2009-12-01

申请号：US11130492

申请日：2005-05-17

申请人： Ronald D. Paul ,  James L. Wyatt ,  Barbara Carrillo ,  Oilda Rubio ,  Diana B. Careaga ,  Lidice L. Lopez

发明人： Ronald D. Paul ,  James L. Wyatt ,  Barbara Carrillo ,  Oilda Rubio ,  Diana B. Careaga ,  Lidice L. Lopez

IPC分类号： G01N33/567

CPC分类号： G01N33/56966 ,  G01N15/1459 ,  G01N33/56972 ,  G01N2015/1006 ,  G01N2015/1402 ,  Y10T436/101666 ,  Y10T436/107497 ,  Y10T436/13 ,  Y10T436/25125

摘要： A method useful for the enumeration of cell populations in a biological sample includes the steps of reacting in a single reaction mixture a sample, a first antibody labeled with a fluorochrome having a first emission spectrum and an additional antibody. The first antibody binds to an antigenic determinant differentially expressed on leukocytes and non-leukocytes. The additional antibody binds to an antigenic determinant differentially expressed on mature and immature granulocytes or myeloid cells, and is labeled either with the first fluorochrome or an additional fluorochrome having an emission spectrum distinguishable from the first emission spectrum. The reaction mixture can be mixed with a nucleic acid dye having an emission spectrum that overlaps with one of the first or additional emission spectra. The reaction mixture may be treated with a lytic system that differentially lyses non-nucleated red blood cells and conserves leukocytes. Populations of hematological cells are detected and enumerated using at least two parameters (fluorescence, optical, and electrical) for each population.

摘要翻译： 可用于计数生物样品中细胞群体的方法包括以下步骤：在单一反应混合物中使样品，用具有第一发射光谱的荧光染料标记的第一抗体和另外的抗体反应。 第一抗体结合在白细胞和非白细胞上差异表达的抗原决定簇。 另外的抗体结合在成熟和未成熟粒细胞或骨髓细胞上差异表达的抗原决定簇，并且用第一荧光染料或具有与第一发射光谱可区分的发射光谱的附加荧光染料进行标记。 反应混合物可以与具有与第一或另外的发射光谱之一重叠的发射光谱的核酸染料混合。 反应混合物可以用裂解体系来处理，所述裂解系统差异地裂解非成核红细胞并保存白细胞。 使用每个群体的至少两个参数（荧光，光学和电）检测和计数血细胞的群体。

公开(公告)号：US07674598B2

公开(公告)日：2010-03-09

申请号：US11475313

申请日：2006-06-27

申请人： Ronald D. Paul ,  Oilda Rubio ,  Diana B. Careaga ,  Lidice L. Lopez ,  Ravindra Mylvaganam

发明人： Ronald D. Paul ,  Oilda Rubio ,  Diana B. Careaga ,  Lidice L. Lopez ,  Ravindra Mylvaganam

IPC分类号： G01N33/567

CPC分类号： G01N33/56966 ,  G01N15/1459 ,  G01N33/56972 ,  G01N2015/1006 ,  G01N2015/1402 ,  Y10T436/101666 ,  Y10T436/107497 ,  Y10T436/13 ,  Y10T436/25125

摘要： Methods for differentially identifying cells in an instrument employ compositions containing a combination of selected antibodies and fluorescent dyes having different cellular distribution patterns and specificities, as well as antibodies and fluorescent dyes characterized by overlapping emission spectra which form non-compensatable spectral patterns. When utilizing the compositions described herein consisting of fluorescent dyes and fluorochrome labeled antibodies with overlapping spectra that cannot be separated or distinguished based upon optical or electronic compensation means, a new fluorescent footprint is established. This new fluorescent footprint is a result of the overlapping spectra and the combined cellular staining patterns of the dyes and fluorochrome labeled antibodies chosen for the composition. The new fluorescent footprint results in histogram patterns that are useful for the identification of additional cell populations or subtypes in hematological disease.

摘要翻译： 用于差异鉴定仪器中的细胞的方法使用含有选择的抗体和具有不同细胞分布模式和特异性的荧光染料的组合的组合物，以及以形成不可补偿光谱图案的重叠发射光谱为特征的抗体和荧光染料。 当利用由本文描述的由荧光染料和荧光染料标记的抗体组成的组合物时，基于光学或电子补偿装置不能被分离或区分的重叠光谱，建立了新的荧光足迹。 这种新的荧光足迹是由组合物选择的染料和荧光染料标记的抗体的重叠光谱和组合的细胞染色模式的结果。 新的荧光足迹导致直方图模式，可用于鉴定血液疾病中的其他细胞群体或亚型。

公开(公告)号：US20050260766A1

公开(公告)日：2005-11-24

申请号：US11130492

申请日：2005-05-17

申请人： Ronald Paul ,  James Wyatt ,  Barbara Carrillo ,  Oilda Rubio ,  Diana Careaga ,  Lidice Lopez

发明人： Ronald Paul ,  James Wyatt ,  Barbara Carrillo ,  Oilda Rubio ,  Diana Careaga ,  Lidice Lopez

IPC分类号： G01N33/53 ,  G01N33/569

CPC分类号： G01N33/56966 ,  G01N15/1459 ,  G01N33/56972 ,  G01N2015/1006 ,  G01N2015/1402 ,  Y10T436/101666 ,  Y10T436/107497 ,  Y10T436/13 ,  Y10T436/25125

摘要： A method useful for the enumeration of cell populations in a biological sample includes the steps of reacting in a single reaction mixture a sample, a first antibody labeled with a fluorochrome having a first emission spectrum and an additional antibody. The first antibody binds to an antigenic determinant differentially expressed on leukocytes and non-leukocytes. The additional antibody binds to an antigenic determinant differentially expressed on mature and immature granulocytes or myeloid cells, and is labeled either with the first fluorochrome or an additional fluorochrome having an emission spectrum distinguishable from the first emission spectrum. The reaction mixture can be mixed with a nucleic acid dye having an emission spectrum that overlaps with one of the first or additional emission spectra. The reaction mixture may be treated with a lytic system that differentially lyses non-nucleated red blood cells and conserves leukocytes. Populations of hematological cells are detected and enumerated using at least two parameters (fluorescence, optical, and electrical) for each population.

摘要翻译： 可用于计数生物样品中细胞群体的方法包括以下步骤：在单一反应混合物中使样品，用具有第一发射光谱的荧光染料标记的第一抗体和另外的抗体反应。 第一抗体结合在白细胞和非白细胞上差异表达的抗原决定簇。 另外的抗体结合在成熟和未成熟粒细胞或骨髓细胞上差异表达的抗原决定簇，并且用第一荧光染料或具有与第一发射光谱可区分的发射光谱的附加荧光染料进行标记。 反应混合物可以与具有与第一或另外的发射光谱之一重叠的发射光谱的核酸染料混合。 反应混合物可以用裂解体系来处理，所述裂解系统差异地裂解非成核红细胞并保存白细胞。 使用每个群体的至少两个参数（荧光，光学和电）检测和计数血细胞的群体。

公开(公告)号：US20060269970A1

公开(公告)日：2006-11-30

申请号：US11475313

申请日：2006-06-27

申请人： Ronald Paul ,  Oilda Rubio ,  Diana Careaga ,  Lidice Lopez ,  Ravindra Mylvaganam

发明人： Ronald Paul ,  Oilda Rubio ,  Diana Careaga ,  Lidice Lopez ,  Ravindra Mylvaganam

IPC分类号： G01N33/567 ,  C07K16/46

CPC分类号： G01N33/56966 ,  G01N15/1459 ,  G01N33/56972 ,  G01N2015/1006 ,  G01N2015/1402 ,  Y10T436/101666 ,  Y10T436/107497 ,  Y10T436/13 ,  Y10T436/25125

摘要： Methods for differentially identifying cells in an instrument employ compositions containing a combination of selected antibodies and fluorescent dyes having different cellular distribution patterns and specificities, as well as antibodies and fluorescent dyes characterized by overlapping emission spectra which form non-compensatable spectral patterns. When utilizing the compositions described herein consisting of fluorescent dyes and fluorochrome labeled antibodies with overlapping spectra that cannot be separated or distinguished based upon optical or electronic compensation means, a new fluorescent footprint is established. This new fluorescent footprint is a result of the overlapping spectra and the combined cellular staining patterns of the dyes and fluorochrome labeled antibodies chosen for the composition. The new fluorescent footprint results in histogram patterns that are useful for the identification of additional cell populations or subtypes in hematological disease.

摘要翻译： 用于差异鉴定仪器中的细胞的方法使用含有选择的抗体和具有不同细胞分布模式和特异性的荧光染料的组合的组合物，以及以形成不可补偿光谱图案的重叠发射光谱为特征的抗体和荧光染料。 当利用由本文描述的由荧光染料和荧光染料标记的抗体组成的组合物时，基于光学或电子补偿装置不能被分离或区分的重叠光谱，建立了新的荧光足迹。 这种新的荧光足迹是由组合物选择的染料和荧光染料标记的抗体的重叠光谱和组合的细胞染色模式的结果。 新的荧光足迹导致直方图模式，可用于鉴定血液疾病中的其他细胞群体或亚型。

公开(公告)号：US07354773B2

公开(公告)日：2008-04-08

申请号：US10437695

申请日：2003-05-14

申请人： Oilda Rubio ,  Carlos Aparicio ,  John A. Maples ,  Julie Wilkinson ,  Cecilia Smith ,  Frank J. Lucas

发明人： Oilda Rubio ,  Carlos Aparicio ,  John A. Maples ,  Julie Wilkinson ,  Cecilia Smith ,  Frank J. Lucas

IPC分类号： G01N1/18

CPC分类号： G01N1/30 ,  G01N1/405 ,  G01N15/147 ,  G01N33/5005 ,  G01N33/536 ,  Y10S435/971 ,  Y10T436/107497 ,  Y10T436/111666 ,  Y10T436/113332 ,  Y10T436/25375

摘要： A method and apparatus for preparing biological cell samples for intracellular analysis. The invention is based upon the recognition that many of the steps of the conventional methods for such sample preparation can be eliminated, leading to a process that readily lends itself to automation and the advantages associated therewith. The method of the invention comprises the steps of (a) cell-fixation, (b) permeabilization and (c) staining (or labeling) of intracellular molecules of interest by probes that are readily detectable by flow cytometric techniques, all without any intervening cell-washing (and re-suspension) steps. Rather, the single cell-washing step is effected after these three steps have been carried out. Preferably, the washing step is carried out by passing the fixed, permeabilized and stained cell sample through a semi-permeable membrane that serves to filter out (by transmission) interferants to waste while retaining the cells of interest.

摘要翻译： 一种用于制备用于细胞内分析的生物细胞样品的方法和装置。 本发明基于这样的认识，即可以消除用于这种样品制备的常规方法的许多步骤，导致易于适应自动化的过程以及与其相关的优点。 本发明的方法包括以下步骤：（a）细胞固定，（b）透化和（c）通过流式细胞术技术容易检测的探针染色（或标记）感兴趣的细胞内分子，所有这些都不具有任何中间细胞 洗涤（和重新暂停）步骤。 相反，在进行这三个步骤之后实现单细胞洗涤步骤。 优选地，洗涤步骤通过使固定的，透化的和染色的细胞样品通过半透膜进行，半透膜用于将（通过传输）干燥剂过滤掉，同时保留感兴趣的细胞。