公开(公告)号：US5350695A

公开(公告)日：1994-09-27

申请号：US802593

申请日：1991-12-05

申请人： Gregory M. Colella ,  Daniel Ben-David ,  Albert Cupo ,  Sophie S. Fan ,  Gena Fischer ,  Grace E. Martin ,  Leonard Ornstein

发明人： Gregory M. Colella ,  Daniel Ben-David ,  Albert Cupo ,  Sophie S. Fan ,  Gena Fischer ,  Grace E. Martin ,  Leonard Ornstein

IPC分类号： G01N15/14 ,  G01N1/30 ,  G01N21/27 ,  G01N21/64 ,  G01N33/48 ,  G01N33/487 ,  G01N33/49 ,  G01N33/50 ,  G01N33/72 ,  G01N33/80

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： 表征和区分网织红细胞和成熟红细胞的方法使用试剂组合物，其包括用于染色血液样品中的网织红细胞的有机阳离子染料和用于保持约6至约9的pH的缓冲溶液。染料可以是蓝色吸收染料恶嗪 750或新亚甲基蓝。 当用于网织红细胞和红细胞的等容体积的试剂组合物中包含两性离子表面活性剂时，并且将试剂组合物和全血样品混合物通过流式细胞仪的感测区域，测量每个细胞散射和吸收的光， 红细胞可以与网织红细胞区分开，并从测定的细胞 - 细胞计算出网织红细胞或红细胞的体积，血红蛋白浓度和每种网织红细胞或红细胞的血红蛋白含量，平均细胞体积，平均红细胞血红蛋白浓度和平均细胞血红蛋白， 细胞内体积和血红蛋白浓度。 一种方法还包括调整测量的吸收信号用于伪吸收。

公开(公告)号：US5284771A

公开(公告)日：1994-02-08

申请号：US802674

申请日：1991-12-05

申请人： Sophie S. Fan ,  Daniel Ben-David ,  Albert Cupo ,  Gena Fischer ,  Grace E. Martin ,  Leonard Ornstein ,  Gregory M. Colella

发明人： Sophie S. Fan ,  Daniel Ben-David ,  Albert Cupo ,  Gena Fischer ,  Grace E. Martin ,  Leonard Ornstein ,  Gregory M. Colella

IPC分类号： G01N15/14 ,  G01N21/27 ,  G01N21/64 ,  G01N33/48 ,  G01N33/49 ,  G01N33/50 ,  G01N33/68 ,  G01N33/80 ,  G01N33/96 ,  G01N1/30

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 for staining a subset of cells is reacted with an anticoagulated 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： 使用试剂组合物进行血液样品制备的方法，所述试剂组合物包括两性离子表面活性剂用于细胞的细胞以消除取向噪声。 当包含用于染色细胞子集的试剂组合物与抗凝血液样品反应，并且反应混合物通过流式细胞仪的感测区域时，测量每个细胞散射和吸收的光，染色的细胞可以 与未染色细胞不同，细胞分别为网织红细胞和成熟红细胞时，每个网织红细胞或红细胞的体积和血红蛋白浓度，血红蛋白含量，平均细胞体积，平均红细胞血红蛋白浓度和平均细胞血红蛋白浓度 网织红细胞或红细胞由测定的细胞间细胞体积和血红蛋白浓度计算。

公开(公告)号：US5438003A

公开(公告)日：1995-08-01

申请号：US961591

申请日：1992-10-15

申请人： Gregory M. Colella ,  Daniel Ben-David ,  Albert Cupo ,  Sophie S. Fan ,  Gena Fischer ,  Grace E. Martin ,  Leonard Ornstein

发明人： Gregory M. Colella ,  Daniel Ben-David ,  Albert Cupo ,  Sophie S. Fan ,  Gena Fischer ,  Grace E. Martin ,  Leonard Ornstein

IPC分类号： G01N15/14 ,  G01N1/30 ,  G01N21/27 ,  G01N21/64 ,  G01N33/48 ,  G01N33/487 ,  G01N33/49 ,  G01N33/50 ,  G01N33/72 ,  G01N33/80

CPC分类号： G01N33/80 ,  G01N1/30 ,  G01N33/5094 ,  Y10S436/80 ,  Y10T436/10 ,  Y10T436/101666 ,  Y10T436/107497

摘要： A reagent composition which includes organic cationic dyes for staining the reticulocytes in a blood sample and buffer solutions for maintaining a pH of about 6 to about 9 is provided. The dyes may be the blue absorption dyes Oxazine 750 or New Methylene Blue. A zwitterionic surfactant is included in the reagent composition for isovolumetric sphering of the reticulocytes and erythrocytes. The reagent composition and whole blood sample mixture are 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.

摘要翻译： 提供了一种试剂组合物，其包括用于染色血液样品中的网织红细胞的有机阳离子染料和用于保持约6至约9的pH的缓冲溶液。 染料可以是蓝色吸收染料Oxazine 750或New Methylene Blue。 用于网织红细胞和红细胞的等容体积的试剂组合物中包含两性离子表面活性剂。 试剂组合物和全血样品混合物通过流式细胞仪的感测区域。 测量每个电池散射和吸收的光; 红细胞可以与网织红细胞区分开，并从测定的细胞 - 细胞计算出网织红细胞或红细胞的体积，血红蛋白浓度和每种网织红细胞或红细胞的血红蛋白含量，平均细胞体积，平均红细胞血红蛋白浓度和平均细胞血红蛋白， 细胞内体积和血红蛋白浓度。

公开(公告)号：US5633167A

公开(公告)日：1997-05-27

申请号：US362744

申请日：1994-12-22

申请人： Sophie S. Fan ,  Daniel Ben-David ,  Albert Cupo ,  Gena Fischer ,  Grace E. Martin ,  Leonard Ornstein ,  Gregory M. Colella

发明人： Sophie S. Fan ,  Daniel Ben-David ,  Albert Cupo ,  Gena Fischer ,  Grace E. Martin ,  Leonard Ornstein ,  Gregory M. Colella

IPC分类号： G01N15/14 ,  G01N21/27 ,  G01N21/64 ,  G01N33/48 ,  G01N33/49 ,  G01N33/50 ,  G01N33/68 ,  G01N33/80 ,  G01N33/96

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： 使用试剂组合物进行血液样品制备的方法，所述试剂组合物包括两性离子表面活性剂用于细胞的细胞以消除取向噪声。 当包含用于染色细胞子集的染料的试剂组合物与血液样品反应，并且反应混合物通过流式细胞仪的感测区域时，测量每个细胞散射和吸收的光，染色的细胞可以 与未染色细胞不同，细胞分别为网织红细胞和成熟红细胞时，每个网织红细胞或红细胞的体积和血红蛋白浓度，血红蛋白含量，平均细胞体积，平均红细胞血红蛋白浓度和平均细胞血红蛋白浓度 网织红细胞或红细胞由测定的细胞间细胞体积和血红蛋白浓度计算。

公开(公告)号：US5360739A

公开(公告)日：1994-11-01

申请号：US802585

申请日：1991-12-05

申请人： Sophie S. Fan ,  Daniel Ben-David ,  Gregory M. Colella ,  Albert Cupo ,  Gena Fischer ,  Leonard Ornstein

发明人： Sophie S. Fan ,  Daniel Ben-David ,  Gregory M. Colella ,  Albert Cupo ,  Gena Fischer ,  Leonard Ornstein

IPC分类号： G01N1/30 ,  G01N15/14 ,  G01N21/27 ,  G01N21/64 ,  G01N33/48 ,  G01N33/487 ,  G01N33/49 ,  G01N33/50 ,  G01N33/72 ,  G01N33/80

CPC分类号： G01N33/80 ,  G01N1/30 ,  G01N15/1459 ,  G01N33/5094 ,  G01N1/38 ,  G01N15/1463 ,  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： 表征和区分网织红细胞和成熟红细胞的方法使用试剂组合物，其包括用于染色血液样品中的网织红细胞的有机阳离子染料和用于保持约6至约9的pH的缓冲溶液。染料可以是红色可激发 荧光染料Oxazine 750，或蓝色可激发荧光染料吖啶橙或吖啶橙衍生物。 当用于网织红细胞和红细胞的等容体积的试剂组合物中包含两性离子表面活性剂时，试剂组合物和全血样品混合物通过流式细胞计数器的感测区域时，散射在至少一个角度间隔的光， 测量每个细胞发荧光的红细胞，网织红细胞和网织红细胞或红细胞的体积，血红蛋白浓度和血红蛋白含量以及网织红细胞的平均细胞体积，平均红细胞血红蛋白浓度和平均细胞血红蛋白和/ 或红细胞从测量的细胞逐个体积和血红蛋白浓度计算。

公开(公告)号：US5411891A

公开(公告)日：1995-05-02

申请号：US961582

申请日：1992-10-15

申请人： Sophie S. Fan ,  Daniel Ben-David ,  Gregory M. Colella ,  Albert Cupo ,  Gena Fischer ,  Leonard Ornstein

发明人： Sophie S. Fan ,  Daniel Ben-David ,  Gregory M. Colella ,  Albert Cupo ,  Gena Fischer ,  Leonard Ornstein

IPC分类号： G01N1/30 ,  G01N15/14 ,  G01N21/27 ,  G01N21/64 ,  G01N33/48 ,  G01N33/487 ,  G01N33/49 ,  G01N33/50 ,  G01N33/72 ,  G01N33/80

CPC分类号： G01N33/80 ,  G01N1/30 ,  G01N15/1459 ,  G01N33/5094 ,  G01N1/38 ,  G01N15/1463 ,  G01N2015/0076 ,  G01N2015/1486 ,  Y10S436/80 ,  Y10T436/10 ,  Y10T436/101666

摘要： A reagent composition 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 is provided. The dyes may be the red excitable fluorescent dye Oxazine 750, or the blue excitable fluorescent dyes Acridine Orange or derivatives of Acridine Orange. A zwitterionic surfactant is included in the reagent composition for isovolumetric sphering of the reticulocytes and erythrocytes. 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： 提供了一种试剂组合物，其包括用于染色血液样品中的网织红细胞的有机阳离子染料和用于保持约6至约9的pH的缓冲溶液。 染料可以是红色可激发荧光染料Oxazine 750，或蓝色可激发荧光染料吖啶橙或吖啶橙衍生物。 用于网织红细胞和红细胞的等容体积的试剂组合物中包含两性离子表面活性剂。 试剂组合物和全血样品混合物通过流式细胞仪的感测区域，测量散布在至少一个角度间隔内并由每个细胞发荧光的光，红细胞可与网织红细胞和体积，血红蛋白浓度 并根据测定的细胞间细胞体积和血红蛋白浓度计算网织红细胞或红细胞的血红蛋白含量，以及网织红细胞和/或红细胞的平均细胞体积，平均红细胞血红蛋白浓度和平均细胞血红蛋白。

公开(公告)号：US07670798B2

公开(公告)日：2010-03-02

申请号：US10442706

申请日：2003-05-20

申请人： Leonard Ornstein ,  Gena Fischer ,  David Zelmanovic ,  Pamela Elsins ,  Jolanta Kunicka ,  Michael J. Malin

发明人： Leonard Ornstein ,  Gena Fischer ,  David Zelmanovic ,  Pamela Elsins ,  Jolanta Kunicka ,  Michael J. Malin

IPC分类号： G01N1/00 ,  G01N7/00 ,  G01N31/00 ,  G01N33/48 ,  G01N33/546

CPC分类号： G01N33/96 ,  A01N1/00 ,  G01N15/1459 ,  G01N2015/0073 ,  G01N2015/1486 ,  Y10T436/10 ,  Y10T436/11 ,  Y10T436/2525

摘要： The present invention describes semi- and fully-automated methods and reagents therefor for the assay and analysis of body fluid samples, particularly non-blood samples. The methods and reagents are especially useful for the assay and analysis of cerebrospinal fluid (CSF) samples. The reagent compositions sphere and fix all cells in the sample in suspension. Reported results can include red blood cell (RBC) and white blood cell (WBC) counts, WBC differential values, cell-by-cell volumes and dry-mass concentrations.

摘要翻译： 本发明描述了用于体液样品，特别是非血液样品的测定和分析的半自动和全自动化方法和试剂。 方法和试剂对于脑脊液（CSF）样品的测定和分析特别有用。 试剂组合物球体并将悬浮液中的样品中的所有细胞固定。 报告的结果可以包括红细胞（RBC）和白血球（WBC）计数，WBC差异值，逐个细胞体积和干质量浓度。

公开(公告)号：US20050026240A1

公开(公告)日：2005-02-03

申请号：US10896338

申请日：2004-07-21

申请人： Michael Sommer ,  Pasquale Degiorgio ,  Bronislaw Czech ,  Gena Fischer ,  E. Chapman ,  David Zelmanovic ,  Jolanta Kunicka

发明人： Michael Sommer ,  Pasquale Degiorgio ,  Bronislaw Czech ,  Gena Fischer ,  E. Chapman ,  David Zelmanovic ,  Jolanta Kunicka

IPC分类号： G01N33/49 ,  G01N1/30 ,  G01N15/14 ,  G01N33/48 ,  G01N33/50

CPC分类号： G01N33/5002 ,  G01N2015/1486 ,  G01N2015/1488 ,  Y10T436/101666 ,  Y10T436/107497

摘要： The enumeration and analysis of residual white blood cells in a sample of leukocyte-reduced blood products is conducted by forming a suspension of the leukocyte-reduced blood products with a sufficient amount of a lysing reagent. The lysing reagent comprises a buffer with a low molar concentration, and a non-ionic surfactant. The suspension of leukocyte-reduced blood products and the lysing reagent is incubated for a sufficient time at a temperature sufficient to selectively lyse the platelets and red blood cells without damaging the white blood cells. The white blood cells of the lysed blood products are then contacted with a suitable dye to stain the white blood cells and the number of stained white blood cells is measured. The lysing reagent is free of harsh organic solvents which can damage the plastic components of automated clinical analyzers.

摘要翻译： 通过用足够量的裂解试剂形成白细胞减少的血液制品的悬浮液来进行白细胞减少的血液制品样品中的残余白细胞的计数和分析。 裂解试剂包含具有低摩尔浓度的缓冲液和非离子表面活性剂。 将白细胞减少的血液制品和裂解试剂的悬浮液在足以在不损伤白细胞的情况下足以选择性地裂解血小板和红细胞的温度下孵育足够的时间。 然后将裂解的血液制品的白细胞与合适的染料接触以染色白细胞，并测量染色的白细胞数。 裂解试剂不含苛刻的有机溶剂，可能会损坏自动化临床分析仪的塑料部件。

公开(公告)号：US5075556A

公开(公告)日：1991-12-24

申请号：US444255

申请日：1989-12-01

申请人： Sophie Fan ,  Gena Fischer

发明人： Sophie Fan ,  Gena Fischer

IPC分类号： G01N21/64 ,  C07D219/08 ,  C09B15/00 ,  G01N15/14 ,  G01N33/48 ,  G01N33/80

CPC分类号： G01N33/80 ,  C09B15/00 ,  Y10T436/145555

摘要： Quaternized derivatives of acridine orange and reagents incorporating such derivatives and their use in quantitatively determining reticulocyte levels in whole blood specimen by fluorescence flow cytometry techniques are disclosed. The quaternized derivatives of acridine orange are of the general formula: ##STR1## wherein Y is bromide (Br.sup.-) or iodide (I.sup.-), andX may be R.sub.1 and/or R.sub.2 substituted benzyl group ##STR2## R.sub.1 is hydrogen or fluorine, and R.sub.2 is fluorine, trifluoromethyl or hydrogen, or X may be hydroxyl ethylene.

公开(公告)号：US07745219B2

公开(公告)日：2010-06-29

申请号：US10896338

申请日：2004-07-21

申请人： Michael J. Sommer ,  Pasquale Degiorgio ,  Bronislaw P. Czech ,  Gena Fischer ,  E. Sabrinah Chapman ,  David Zelmanovic ,  Jolanta Kunicka

发明人： Michael J. Sommer ,  Pasquale Degiorgio ,  Bronislaw P. Czech ,  Gena Fischer ,  E. Sabrinah Chapman ,  David Zelmanovic ,  Jolanta Kunicka

IPC分类号： G01N33/48 ,  G01N33/49

CPC分类号： G01N33/5002 ,  G01N2015/1486 ,  G01N2015/1488 ,  Y10T436/101666 ,  Y10T436/107497

摘要： The enumeration and analysis of residual white blood cells in a sample of leukocyte-reduced blood products is conducted by forming a suspension of the leukocyte-reduced blood products with a sufficient amount of a lysing reagent. The lysing reagent comprises a buffer with a low molar concentration, and a non-ionic surfactant. The suspension of leukocyte-reduced blood products and the lysing reagent is incubated for a sufficient time at a temperature sufficient to selectively lyse the platelets and red blood cells without damaging the white blood cells. The white blood cells of the lysed blood products are then contacted with a suitable dye to stain the white blood cells and the number of stained white blood cells is measured. The lysing reagent is free of harsh organic solvents which can damage the plastic components of automated clinical analyzers.

摘要翻译： 通过用足够量的裂解试剂形成白细胞减少的血液制品的悬浮液来进行白细胞减少的血液制品样品中的残余白细胞的计数和分析。 裂解试剂包含具有低摩尔浓度的缓冲液和非离子表面活性剂。 将白细胞减少的血液制品和裂解试剂的悬浮液在足以在不损伤白细胞的情况下足以选择性地裂解血小板和红细胞的温度下孵育足够的时间。 然后将裂解的血液制品的白细胞与合适的染料接触以染色白细胞，并测量染色的白细胞数。 裂解试剂不含苛刻的有机溶剂，可能会损坏自动化临床分析仪的塑料部件。