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公开(公告)号：EP0170345A3
公开(公告)日：1987-03-25
申请号：EP85302094
申请日：1985-03-26
申请人： TECHNICON INSTRUMENTS CORPORATION (a New York corporation) , THE MOUNT SINAI SCHOOL OF MEDICINE
CPC分类号： G01N33/56972
摘要： A flow cytometry method reproducibly detects and counts a lymphocyte population of interest in a leukocyte suspension or whole blood sample in which the red cells are subsequently lysed. The suspension (or sample) is combined with a reagent comprising a primary antibody, either native, carrying an attached enzyme or biotin or other label, and a fixative reagent, in either order. Where the enzyme is not attached, an enzyme is coupled specifically to the primary antibody. The fixed suspension is reacted with a color- producing enzyme-cytochemical reagent. The suspension, now including stained and unstained fixed cells, is passed through a flow cytometer and the cells are characterized and counted on the basis of their light-scattering and light-absorbing properties.
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2.发明授权Reagent compositions and their use in the identification and characterization of reticulocytes in whole blood 失效试剂组合物及其在识别和使用全血中网织红细胞的表征
公开(公告)号：EP0545315B1
公开(公告)日：1998-01-07
申请号：EP92120309.7
申请日：1992-11-27
发明人： Fan, Sophie S. , Ben-David, Daniel , Collela, Gregory M. , Cupo, Albert , Fischer, Gena , Ornstein, Leonard
CPC分类号： G01N33/80 , G01N1/30 , G01N1/38 , G01N15/1459 , G01N15/1463 , G01N33/5094 , G01N2015/0076 , G01N2015/1486 , Y10S436/80 , Y10T436/10 , Y10T436/101666
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3.发明公开Reagent compositions and their use in the identification and characterization of reticulocytes in whole blood 失效试剂组合物及其在全血中网织红细胞的鉴定和表征使用。
公开(公告)号：EP0545314A1
公开(公告)日：1993-06-09
申请号：EP92120308.9
申请日：1992-11-27
发明人： Colella, Gregory M. , Ben-David, Daniel , Cupo, Albert , Fan, Sophie S. , Fischer, Gena , Martin, Grace E. , Ornstein, Leonard
CPC分类号： G01N33/80 , G01N1/30 , G01N33/5094 , Y10S436/80 , Y10T436/10 , Y10T436/101666 , Y10T436/107497
摘要： Methods for characterizing and distinguishing reticulocytes and mature red blood cells use reagent compositions which include organic cationic dyes for staining the reticulocytes in the blood sample and buffer solutions for maintaining a pH of about 6 to about 9. The dyes may be the blue absorption dyes Oxazine 750 or New Methylene Blue. When a zwitterionic surfactant is included in the reagent composition for isovolumetric sphering of the reticulocytes and erythrocytes, and the reagent composition and whole blood sample mixture is passed through the sensing region of a flow cytometer, the light scattered and absorbed by each cell is measured, the erythrocytes can be distinguished from reticulocytes and the volume, hemoglobin concentration and the hemoglobin content of each reticulocyte or erythrocyte, and the mean cell volume, mean corpuscular hemoglobin concentration, and mean cell hemoglobin of the reticulocytes or erythrocytes are calculated from the measured cell-by-cell volume and hemoglobin concentration. One method also includes adjusting the measured absorption signals for pseudo-absorption.
摘要翻译： 用于表征和区分网织红细胞和成熟红细胞的方法使用的试剂组合物，其包括有机阳离子染料在血液样品和缓冲溶液染色网织红细胞进行的pH为约6维护至约9的染料可以是蓝光吸收染料恶嗪 750新亚甲基蓝。 当两性离子表面活性剂包括在用于网状细胞和红细胞的等容球状，和所述试剂组合物和全血样品混合物中的试剂组合物通过流的感测区流式细胞仪，由每个细胞的散射和吸收的光被测量通过， 红细胞可以从网织红细胞和体积，血红蛋白浓度和每个网织红细胞或红血球，平均细胞体积的血红蛋白含量来区分，平均红细胞血红蛋白浓度，平均网织红细胞或红细胞的细胞血红蛋白是从所测量的小区计算 通过细胞体积和血红蛋白浓度。 因此，一个方法包括调节用于伪吸收测得的吸收信号。
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公开(公告)号：EP0170345A2
公开(公告)日：1986-02-05
申请号：EP85302094.9
申请日：1985-03-26
CPC分类号： G01N33/56972
摘要： A flow cytometry method reproducibly detects and counts a lymphocyte population of interest in a leukocyte suspension or whole blood sample in which the red cells are subsequently lysed. The suspension (or sample) is combined with a reagent comprising a primary antibody, either native, carrying an attached enzyme or biotin or other label, and a fixative reagent, in either order. Where the enzyme is not attached, an enzyme is coupled specifically to the primary antibody. The fixed suspension is reacted with a color- producing enzyme-cytochemical reagent. The suspension, now including stained and unstained fixed cells, is passed through a flow cytometer and the cells are characterized and counted on the basis of their light-scattering and light-absorbing properties.
摘要翻译： 流式细胞术方法可重复地检测并计数随后裂解红细胞的白细胞悬浮液或全血样品中感兴趣的淋巴细胞群。 将悬浮液（或样品）与包含天然的，携带附着的酶或生物素或其它标记的一级抗体和固定剂的试剂以任何顺序组合。 当酶不连接时，酶与第一抗体特异性地偶联。 将固定的悬浮液与产生色素的酶 - 细胞化学试剂反应。 将包含染色和未染色的固定细胞的悬浮液通过流式细胞仪，并根据其光散射和光吸收特性对细胞进行表征和计数。
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公开(公告)号：EP0545313B1
公开(公告)日：1997-07-23
申请号：EP92120307.1
申请日：1992-11-27
发明人： Fan, Sophie S. , Ben-David, Daniel , Cupo, Albert , Fischer, Gena , Martin, Grace E. , Ornstein, Leonard , Colella, Gregory M.
CPC分类号： G01N33/5002 , G01N15/1459 , G01N33/80 , G01N33/96 , G01N2015/1402 , G01N2015/1477 , Y10T436/10 , Y10T436/101666 , Y10T436/102499 , Y10T436/107497 , Y10T436/108331
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6.发明公开Reagent compositions and their use in the identification and characterization of reticulocytes in whole blood 失效试剂组合物及其在全血中网织红细胞的鉴定和表征使用。
公开(公告)号：EP0545315A1
公开(公告)日：1993-06-09
申请号：EP92120309.7
申请日：1992-11-27
发明人： Fan, Sophie S. , Ben-David, Daniel , Collela, Gregory M. , Cupo, Albert , Fischer, Gena , Ornstein, Leonard
CPC分类号： G01N33/80 , G01N1/30 , G01N1/38 , G01N15/1459 , G01N15/1463 , G01N33/5094 , G01N2015/0076 , G01N2015/1486 , Y10S436/80 , Y10T436/10 , Y10T436/101666
摘要： Methods for characterizing and distinguishing reticulocytes and mature red blood cells use reagent compositions which include an organic cationic dye for staining the reticulocytes in the blood sample and a buffer solution for maintaining a pH of about 6 to about 9. The dyes may be the red excitable fluorescent dye Oxazine 750, or the blue excitable fluorescent dyes Acridine Orange or derivatives of Acridine Orange. When a zwitterionic surfactant is included in the reagent composition for isovolumetric sphering of the reticulocytes and erythrocytes, and the reagent composition and whole blood sample mixture is passed through the sensing region of a flow cytometer, the light scattered through at least one angular interval and that fluoresced by each cell is measured, the erythrocytes can be distinguished from reticulocytes and the volume, hemoglobin concentration and the hemoglobin content of each reticulocyte or erythrocyte, and the mean cell volume, mean corpuscular hemoglobin concentration, and mean cell hemoglobin of the reticulocytes and/or erythrocytes are calculated from the measured cell-by-cell volume and hemoglobin concentration.
摘要翻译： 用于表征和区分网织红细胞和成熟红细胞的方法使用的试剂组合物，其包括在有机阳离子染料染色的血液样品和为pH约6维护至约9的染料的缓冲液在网织红细胞可以是红色激发的 荧光染料恶嗪750，或蓝色激发的荧光染料吖啶橙或吖啶橙的衍生物。 当两性离子表面活性剂包括在用于网状细胞和红细胞的等容球状，和所述试剂组合物和全血样品混合物中的试剂组合物通过流的感测区流式细胞仪通过，光散射通过至少一个角度间隔，做 由每个细胞发出的荧光进行测量，红细胞可以从网织红细胞区分并且音量，血红蛋白浓度和每个网织红细胞或红血球，平均细胞体积的血红蛋白含量，平均红细胞血红蛋白浓度，平均网织红细胞和细胞血红蛋白/ 或红细胞是从所测量的小区由小区体积和血红蛋白浓度计算。
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公开(公告)号：EP0545313A1
公开(公告)日：1993-06-09
申请号：EP92120307.1
申请日：1992-11-27
发明人： Fan, Sophie S. , Ben-David, Daniel , Cupo, Albert , Fischer, Gena , Martin, Grace E. , Ornstein, Leonard , Colella, Gregory M.
CPC分类号： G01N33/5002 , G01N15/1459 , G01N33/80 , G01N33/96 , G01N2015/1402 , G01N2015/1477 , Y10T436/10 , Y10T436/101666 , Y10T436/102499 , Y10T436/107497 , Y10T436/108331
摘要： Methods for blood sample preparation using reagent compositions which include a zwitterionic surfactant for sphering of cells to eliminate orientational noise. When the reagent composition including a dye for staining a subset of cells is reacted with a blood sample, and the reaction mixture is passed through the sensing region of a flow cytometer, the light scattered and absorbed by each cell is measured, the stained cells can be distinguished from the unstained cells, and when the cells are reticulocytes and mature erythrocytes, respectively, the volume and hemoglobin concentration of each reticulocyte or erythrocyte, and the hemoglobin content, mean cell volume, mean corpuscular hemoglobin concentration, and mean cell hemoglobin of the reticulocytes or erythrocytes are calculated from the measured cell-by-cell volume and hemoglobin concentration.
摘要翻译： 使用的试剂组合物用于血液样品制备方法包括用于细胞的球形两性离子表面以消除取向噪声。 当试剂组合物包括对染色细胞的一个子集进行反应与血样的染料，并且将反应混合物通过流的感测区流式细胞仪通过，由每个细胞的散射和吸收的光进行测量，将染色的细胞可以 从未染色的细胞区分开来，并且当所述细胞是网织红细胞和成熟红细胞，分别各网织红细胞或红细胞和血红蛋白含量，平均细胞体积的体积和血红蛋白浓度，平均红细胞血红蛋白浓度，平均的细胞血红蛋白 网织红细胞或红细胞是从所测量的小区由小区体积和血红蛋白浓度计算。
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8.发明授权Reagent compositions and their use in the identification and characterization of reticulocytes in whole blood 失效试剂组合物及其在识别和使用全血中网织红细胞的表征
公开(公告)号：EP0545314B1
公开(公告)日：1998-01-07
申请号：EP92120308.9
申请日：1992-11-27
发明人： Colella, Gregory M. , Ben-David, Daniel , Cupo, Albert , Fan, Sophie S. , Fischer, Gena , Martin, Grace E. , Ornstein, Leonard
CPC分类号： G01N33/80 , G01N1/30 , G01N33/5094 , Y10S436/80 , Y10T436/10 , Y10T436/101666 , Y10T436/107497
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公开(公告)号：EP0154522A3
公开(公告)日：1988-11-02
申请号：EP85301390
申请日：1985-02-28
发明人： Ornstein, Leonard , Kim, Young Ran
CPC分类号： G01N33/48 , Y10T436/101666 , Y10T436/107497
摘要： A one-step method (and sphering agent/fixing agent useful therein) for treating mammalian red blood cells in a sample which can be effectively measured electrooptically for determination of red blood cell volumes, which comprises treating an anticoagulated whole blood sample with a single aqueous sphering agent/fixing agent reagent at a temperature in the range of 4°C to 37°C over a reaction period of up to 180 seconds employing a whole blood dilution factor of from 1:300 to 1:4,000 wherein said ona step, single reagent treatment provides isovoluments sphering of the erythrocytes in the semple independent of the concentration of said erythrocytes in the whole Diood sample and without inducing lysis of said erychwocytes in the resulting diluted semple. The preferred sptering agent is an alkali metal salt of an alkyl sulfate irr an amount 0.5 to 3.0 mg %, and the preferred fixing agent is an aldehyde in an amount from 0.05 to 0.3%.
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