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公开(公告)号：US06911313B2
公开(公告)日：2005-06-28
申请号：US09058323
申请日：1998-04-09
IPC分类号： G01N33/50 , G01N33/569 , G01N33/80 , G01N33/53 , G01N21/76
CPC分类号： G01N33/5094 , G01N15/1459 , G01N33/56972 , G01N33/80 , G01N2015/1006 , G01N2015/1402 , Y10T436/107497 , Y10T436/108331 , Y10T436/13 , Y10T436/25125 , Y10T436/2525 , Y10T436/25375
摘要： A method for discriminating and counting erythroblasts comprises the steps of: (i) staining leukocytes in a hematologic sample by adding a fluorescent labeled antibody capable of binding specifically with leukocytes to the hematologic sample; (ii) raising the permeability only of cell membranes of erythroblasts in the hematologic sample to a nucleotide fluorescent dye which does not permeate a cell membrane usually, the nucleotide fluorescent dye having a fluorescent spectrum capable of being distinguished from that of a fluorescent labeling compound of the fluorescent labeled antibody in step (i); (iii) staining nuclei of the erythroblasts in the hematologic sample with the nucleotide fluorescent dye; (iv) subjecting the hematologic sample to flowcytometry to detect at least two fluorescent signals from each cell; and (v) discriminating and counting the erythroblasts from difference in intensity between the at least two fluorescent signals.
摘要翻译： 用于鉴别和计数红细胞的方法包括以下步骤：（i）通过将能够与白细胞特异性结合的荧光标记抗体加入到血液样品中来染色血液学样品中的白细胞; （ii）将血液学样品中的红细胞的细胞膜的渗透性提高至不透过细胞膜的核苷酸荧光染料，所述核苷酸荧光染料具有与荧光标记化合物的荧光标记化合物不同的荧光光谱 步骤（i）中的荧光标记抗体; （iii）用核苷酸荧光染料染色血液样品中的红细胞核; （iv）使血液样品经流式细胞术检测来自每个细胞的至少两个荧光信号; 和（v）鉴别和计数红细胞与至少两个荧光信号之间的强度差异。
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公开(公告)号：US06900023B1
公开(公告)日：2005-05-31
申请号：US09388899
申请日：1999-09-02
IPC分类号： G01N33/49 , G01N15/14 , G01N33/50 , G01N33/53 , G01N33/536 , G01N33/569
CPC分类号： G01N33/56972 , G01N33/5094 , G01N2333/70546 , Y10S435/962 , Y10S436/80 , Y10S436/824 , Y10S436/825 , Y10T436/101666 , Y10T436/107497 , Y10T436/13 , Y10T436/25 , Y10T436/25125
摘要： A method for classifying and counting leukocytes comprises the steps of: (1) adding to a hematological sample the following fluorescence-labeled antibodies labeled with fluorescent dyes which emit fluorescences distinguishable from each other; (a) a first fluorescence-labeled antibody (1st antibody) which bonds specifically to leukocytes, (b) a second fluorescence-labeled antibody (2nd antibody) which bonds to at least one kind of neutrophilic cells, and (c) a third fluorescence-labeled antibody (3rd antibody) which bonds to at least one kind of immature granulocytic cells, in order to stain leukocytic cells in the sample, and removing erythrocytes from the sample; (2) analyzing the resulting sample using a flow cytometer to measure at least one scattered light signal and three separate fluorescence signals; (3) defining a group of granulocytic cells on the basis of intensity of the scattered light and intensity of fluorescence from the 1st antibody; (4) defining neutrophilic cells in the defined group of granulocylic cells on the basis of the intensity of the fluorescence from the 1st antibody and intensity of fluorescence from the 2nd or 3rd antibody; (5) classifying the defined group of the neutrophilic cells into groups of neutrophilic cells different in degree of maturity on the basis of the intensity of the fluorescence from the 2nd antibody and the intensity of the fluorescence from the 3rd antibody, and counting the number of cells in each of the groups.
摘要翻译： 用于白细胞分类和计数的方法包括以下步骤：（1）向血液样品中加入以荧光染料标记的荧光标记的抗体，所述荧光染料发射荧光彼此区分; （a）与白细胞特异性结合的第一荧光标记抗体（第一抗体），（b）与至少一种嗜中性粒细胞结合的第二荧光标记抗体（第二抗体），和（c）第三荧光 标记的抗体（第3抗体），其与至少一种未成熟的粒细胞细胞结合，以染色样品中的白细胞，并从样品中除去红细胞; （2）使用流式细胞仪分析所得样品以测量至少一个散射光信号和三个分离的荧光信号; （3）基于散射光的强度和第一抗体的荧光强度来定义一组粒细胞; （4）基于来自第一抗体的荧光强度和来自第二或第三抗体的荧光强度来定义所定义的颗粒细胞组中的嗜中性细胞; （5）基于来自第二抗体的荧光强度和来自第三抗体的荧光强度，将所定义的嗜中性粒细胞群分类成不同程度的嗜中性粒细胞组，并计数第 每个组中的细胞。
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公开(公告)号：USRE39006E1
公开(公告)日：2006-03-07
申请号：US10704463
申请日：2003-11-07
申请人： Berend Houwen , Yukio Tsujino , Takashi Morikawa , Yoshiro Ikeuchi , Yukio Hamaguchi , Fu-sheng Wang , Kojiro Hirai
发明人： Berend Houwen , Yukio Tsujino , Takashi Morikawa , Yoshiro Ikeuchi , Yukio Hamaguchi , Fu-sheng Wang , Kojiro Hirai
IPC分类号： C12Q1/04
CPC分类号： G01N33/5094
摘要： Improved method for detecting and/or counting the appearance of hematopoietic progenitor cells which comprises the steps of mixing a blood sample with a reagent capable of detecting immature cells without employing any immunological techniques, obtaining cell information about the thus treated sample using a particle analyzer and constructing a cell distribution profile, delineating a portion of the profile as a zone in which at least one subclass of hematopoietic progenitor cells appear, and counting the cells within the zone. The method is capable of detecting the appearance of hematopoietic progenitor cells in a simple and positive manner.
摘要翻译： 用于检测和/或计数造血祖细胞外观的改进方法，其包括以下步骤：将血液样品与能够检测未成熟细胞的试剂混合而不使用任何免疫技术，使用粒子分析仪获得关于如此处理的样品的细胞信息; 构建细胞分布轮廓，将轮廓的一部分描绘为其中存在至少一个造血祖细胞亚类的区域，并对该区域内的细胞进行计数。 该方法能够以简单和积极的方式检测造血祖细胞的外观。
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公开(公告)号：US07049093B2
公开(公告)日：2006-05-23
申请号：US09992221
申请日：2001-11-06
申请人： Tomohiro Tsuji , Yusuke Mori , Takashi Sakata , Yukio Hamaguchi
发明人： Tomohiro Tsuji , Yusuke Mori , Takashi Sakata , Yukio Hamaguchi
IPC分类号： C12Q1/06
CPC分类号： G01N33/5005 , G01N15/1459 , G01N33/5002 , G01N33/6863 , G01N2015/1006 , G01N2015/1402
摘要： A method for classifying and counting nucleated bone marrow cells comprises the steps of: (1) mixing a sample of bone marrow fluid with an erythrocyte lysing agent to lyse erythrocytes in the sample and render leukocytic cells and erythroid cells in the sample suitable for staining, and staining the sample with a fluorescent dye for producing a difference in intensity of fluorescence between the leukocytic cells and the erythroid cells; (2) introducing the resulting sample to a flow cytometer to detect at least one kind of scattered light and at least one kind of fluorescence; (3) classifying and counting nucleated bone marrow cells, the leukocytic cells and the erythroid cells with use of a difference in the intensity of the fluorescence and the scattered light; (4) calculating the ratio of the nucleated bone marrow cells to the erythroid cells or leukocytic cells from the obtained erythroid cell count or leukocytic cell count and the obtained nucleated bone marrow cell count; and (5) calculating the ratio of the leukocytic cells to the erythroid cells from the erythroid cell count and the leukocytic cell count.
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5.发明授权Diluting reagent and method compelling time-independent consistency in MCV assay 有权稀释试剂和方法在MCV测定中引人注目的时间依赖性一致性公开(公告)号：US06225124B1
公开(公告)日：2001-05-01
申请号：US09323689
申请日：1999-06-02
申请人： Berend Houwen , Kinya Uchihashi , Yukio Hamaguchi , Rolf Mast
发明人： Berend Houwen , Kinya Uchihashi , Yukio Hamaguchi , Rolf Mast
IPC分类号： G01N3348
CPC分类号： G01N33/5094 , G01N15/12 , Y10T436/10 , Y10T436/101666 , Y10T436/108331 , Y10T436/2525 , Y10T436/25625
摘要： Aqueous blood-sample diluting reagent and method of its use for compelling a morphological change in a blood sample to yield an MCV value assayed at elapsed time after the sample is drawn to be consistent within a diagnostically acceptable range with the original, immediate post-drawing MCV value. Selection of a small amount of a predetermined surfactant added within a limited range of concentration, and of a salt for adjusting osmotic pressure of the sample is thereby determined. The blood sample is treated with an anti-coagulant agent immediately post-drawing, and for assay in a particle analyzer at post-drawing elapsed time is diluted with the reagent solution. The reagent has an osmotic pressure (&pgr;) of approximately 150-400 mOsm/kg and a pH of 6.0-8.5. The surfactant is present in a 0.0005% to 0.5% concentration and has a hydrophile-lipophile balance (HLB) of 10-20.
摘要翻译： 水性血液样品稀释试剂及其用于强制血液样品中形态学变化的方法，以产生在样品抽出后经过时间测定的MCV值，使其在诊断上可接受的范围内与原始的立即后绘图 MCV值。 由此确定少量在有限浓度范围内加入的预定表面活性剂和用于调节样品渗透压的盐的选择。 血液样品立即在拉伸后用抗凝剂处理，并且在抽出后经过时间的粒子分析仪中用试剂溶液稀释。 试剂的渗透压（pI）为约150-400mOsm / kg，pH为6.0-8.5。 表面活性剂的浓度为0.0005％至0.5％，亲水亲油平衡（HLB）为10-20。
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公开(公告)号：US5830701A
公开(公告)日：1998-11-03
申请号：US829239
申请日：1997-03-28
IPC分类号： G01N33/48 , C12N5/08 , G01N15/00 , G01N15/14 , G01N33/49 , G01N33/50 , C12Q1/04 , C12Q1/06 , G01N21/76
CPC分类号： G01N33/5094
摘要： Improved method for detecting and/or counting the appearance of hematopoietic progenitor cells which comprises the steps of mixing a blood sample with a reagent capable of detecting immature cells without employing any immunological techniques, obtaining cell information about the thus treated sample using a particle analyzer and constructing a cell distribution profile, delineating a portion of the profile as a zone in which at least one subclass of hematopoietic progenitor cells appear, and counting the cells within the zone. The method is capable of detecting the appearance of hematopoietic progenitor cells in a simple and positive manner.
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公开(公告)号：US20050003471A1
公开(公告)日：2005-01-06
申请号：US10613767
申请日：2003-07-03
申请人： Fu-Sheng Wang , Mark Dorfner , Suma Peesapati , Steve Burger , Kojiro Hirai , Yukio Hamaguchi
发明人： Fu-Sheng Wang , Mark Dorfner , Suma Peesapati , Steve Burger , Kojiro Hirai , Yukio Hamaguchi
IPC分类号： C12Q1/02 , C12R1/91 , G01N1/30 , G01N15/10 , G01N15/12 , G01N15/14 , G01N21/64 , G01N33/49 , G01N33/50 , G01N33/48
CPC分类号： G01N21/47 , G01N1/30 , G01N15/12 , G01N15/147 , G01N21/6428 , G01N33/5094 , G01N2015/1037 , G01N2015/105 , G01N2015/1087
摘要： Methods of detecting megakaryocytes are described that include (a) providing a sample that contains a cell; (b) detecting a plurality of morphological information from the cell; (c) generating a scattergram from the plurality of morphological information; and (d) determining whether a population exists in a megakaryocyte region of the scattergram.
摘要翻译： 描述了检测巨核细胞的方法，其包括（a）提供含有细胞的样品; （b）从小区检测多个形态信息; （c）从所述多个形态信息生成散点图; 和（d）确定群体是否存在于散点图的巨核细胞区域中。
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8.发明授权Reagent for simultaneous determination of hemoglobin and leukocytes in blood 失效用于同时测定血液中血红蛋白和白细胞的试剂
公开(公告)号：US5250437A
公开(公告)日：1993-10-05
申请号：US596207
申请日：1990-10-10
申请人： Syouzou Toda , Takashi Sakata , Yukio Hamaguchi
发明人： Syouzou Toda , Takashi Sakata , Yukio Hamaguchi
CPC分类号： G01N33/721 , G01N33/5094 , Y10T436/10 , Y10T436/101666 , Y10T436/107497 , Y10T436/108331
摘要： A reagent for measuring the hemoglobin concentration and counting the number of leukocytes in a blood sample, which contains at least one of a quaternary ammonium salt or a pyridinium salt, having a concentration capable of hemolyzing erythrocytes in the blood and denaturing hemoglobin and at least one of cationic, nonionic or amphoteric surfactants or an oxidant capable of oxidizing heme in hemoglobin, the reagent being capable of dividing leukocytes into two or three fractions, for example, a fraction of lymphocytes, a fraction of monocytes, eosinophils and basophils, and a fraction of neutrophils.
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9.发明授权
公开(公告)号：US5968832A
公开(公告)日：1999-10-19
申请号：US57250
申请日：1998-04-08
CPC分类号： G01N33/5094 , G01N33/721 , Y10T436/10 , Y10T436/101666 , Y10T436/107497 , Y10T436/108331 , Y10T436/25 , Y10T436/25125 , Y10T436/2525
摘要： A reagent for measurement of leukocytes and hemoglobin concentration in the blood includes a cationic surfactant in an amount sufficient to lyse erythrocytes and denature hemoglobin, at least one of the following hemoglobin stabilizers:(a) sulfosalicylic acid, or its salt, in an amount effective for promoting the conversion of hemoglobin into methemoglobin,(b) 0.2 to 10.0 g/L of a water-soluble chelating agent having a nitrogen atom and a carboxyl group, and(c) piperazine, or its salt, in an amount effective for promoting the conversion of hemoglobin into methemoglobin, anda buffer for maintaining pH at 4 to 6.
摘要翻译： 白血球的测定用试剂和血液中的血红蛋白浓度包括阳离子表面活性剂，其量足以溶解红细胞，变性血红蛋白，以下血红蛋白稳定剂中的至少一种：（a）磺基水杨酸或其盐，有效量 用于促进血红蛋白转化成高铁血红蛋白，（b）0.2〜10.0g / L的具有氮原子和羧基的水溶性螯合剂，和（c）哌嗪或其盐，其量有效促进 将血红蛋白转化成高铁血红蛋白，以及将pH保持在4-6的缓冲液。
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10.再颁专利公开(公告)号：USRE38131E1
公开(公告)日：2003-06-03
申请号：US09977768
申请日：2001-10-15
IPC分类号： G01N3100
CPC分类号： G01N33/5094 , G01N33/721 , Y10T436/10 , Y10T436/101666 , Y10T436/107497 , Y10T436/108331 , Y10T436/25 , Y10T436/25125 , Y10T436/2525
摘要： A reagent for measurement of leukocytes and hemoglobin concentration in the blood includes a cationic surfactant in an amount sufficient to lyse erythrocytes and denature hemoglobin, at least one of the following hemoglobin stabilizers: (a) sulfosalicylic acid, or its salt, in an amount effective for promoting the conversion of hemoglobin into methemoglobin, (b) 0.2 to 10.0 g/L of a water-soluble chelating agent having a nitrogen atom and a carboxyl group, and (c) piperazine, or its salt, in an amount effective for promoting the conversion of hemoglobin into methemoglobin, and a buffer for maintaining pH at 4 to 6.
摘要翻译： 白血球的测定用试剂和血液中的血红蛋白浓度包括阳离子表面活性剂，其量足以溶解红细胞，变性血红蛋白，以下血红蛋白稳定剂中的至少一种：（a）磺基水杨酸或其盐，有效量 用于促进血红蛋白转化成高铁血红蛋白，（b）0.2〜10.0g / L的具有氮原子和羧基的水溶性螯合剂，和（c）哌嗪或其盐，其量有效促进 将血红蛋白转化为高铁血红蛋白，以及将pH保持在4至6的缓冲液。
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