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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：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：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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4.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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公开(公告)号：US5958776A
公开(公告)日：1999-09-28
申请号：US972103
申请日：1997-11-17
CPC分类号： G01N33/5094 , Y10T436/10 , Y10T436/101666 , Y10T436/107497
摘要： A hematological sample is treated with a hemolytic agent that maintains immature leukocytes in a viable state and damages other leukocytes. The damaged leukocytes are stained with a fluorochrome which can stain damaged cells. At least one kind of scattered light and at least one kind of fluorescence of the treated and stained blood cells are measured to classify and count leukocytes based on the intensities of the scattered light and the fluorescence. Highly precise measurement of immature leukocytes and simultaneous classification and counting of normal leukocytes are achieved.
摘要翻译： 用溶血剂处理血液样品，其将未成熟白细胞维持在可行状态并损伤其他白细胞。 损伤的白细胞用可染色受损细胞的荧光染料染色。 根据散射光和荧光的强度，测量处理和染色的血细胞中的至少一种散射光和至少一种荧光，以对白细胞进行分类和计数。 实现了对未成熟白细胞的高精度测量，并实现了正常白细胞的同时分类和计数。
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6.发明授权Cytoanalyzer using separate sensing portions on a dectector and method for aligning the same 失效在分离器上使用分离的感测部分的细胞分析仪和用于对准其的方法
公开(公告)号：US5737078A
公开(公告)日：1998-04-07
申请号：US512647
申请日：1995-08-08
CPC分类号： G01N15/1459 , G01N15/1434 , G01N2015/008 , G01N2015/1477 , G01N2015/1486 , G01N2021/4716
摘要： A cytoanalyzer includes a flow cell for aligning cells and permitting the same to pass therethrough, a semiconductor light emitting device for emitting a laser beam to the cells passing through the flow cell, and a light-detecting device having at least two separate photosensing portions capable of sensing two types of light scattered by each of the cells. The alignment of the cytoanalyzer may be achieved by comparing intensities of light on the at least two separate photosensing portions.
摘要翻译： 细胞分析仪包括用于对准细胞并允许其通过的流通池，用于向通过流通池的细胞发射激光束的半导体发光装置，以及具有至少两个分开的感光部分的光检测装置 感测由每个单元散射的两种类型的光。 细胞分析仪的对准可以通过比较至少两个单独的感光部分上的光的强度来实现。
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公开(公告)号：US5242832A
公开(公告)日：1993-09-07
申请号：US596205
申请日：1990-10-10
申请人： Takashi Sakata
发明人： Takashi Sakata
CPC分类号： G01N33/5094 , G01N33/721 , Y10T436/10 , Y10T436/101666 , Y10T436/107497 , Y10T436/108331 , Y10T436/2525
摘要： A reagent, satisfying the undermentioned conditions, is useful for counting the number of leukocytes and measuring the hemoglobin concentration in blood samples. The reagent is free of cyanides and it is able to maintain hemoglobin in a stable state for a prolonged time. The reagent contains at least one cationic surfactant selected from the group consisting of a quaternary ammonium salt of a concentration capable of hemolyzing erythrocytes in the blood and oxidizing hemoglobin and a composition of pyridinium salts; at least one cationic, nonionic or amphoteric surfactant; and at least one hemoglobin stabilizer.
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公开(公告)号：US6004816A
公开(公告)日：1999-12-21
申请号：US80016
申请日：1998-05-15
CPC分类号： G01N33/52 , G01N33/5094 , Y10S436/80 , Y10T436/10 , Y10T436/101666 , Y10T436/107497
摘要： A reagent for classification and counting of leukocytes, the reagent containing at least one dye which has the following structural formula ##STR1## where R.sub.1 is a hydrogen atom or an alkyl group, R.sub.2 and R.sub.3 each represent a hydrogen atom, a lower alkyl group or a lower alkoxy group, R.sub.4 represents a hydrogen atom, an acyl group or an alkyl group, Z represents a sulfur atom, an oxygen atom, or a carbon atom having a lower alkyl group, n denotes 0, 1 or 2, and X.sup.- represents an anion,and specifically binds to RNA to increase in fluorescence intensity, and a method for classification and counting of leukocytes which uses the reagent, whereby abnormal cells such as immature leukocytes and abnormal leukocytes can be classified and counted easily and highly accurately, and at the same time, classification and counting of normal leukocytes as well as the counting of leukocytes can be performed.
摘要翻译： 用于白细胞分类和计数的试剂，所述试剂含有至少一种具有以下结构式的染料，其中R 1是氢原子或烷基，R 2和R 3各自表示氢原子，低级烷基或低级烷氧基 基团，R 4表示氢原子，酰基或烷基，Z表示硫原子，氧原子或具有低级烷基的碳原子，n表示0,1或2，X表示阴离子 并特异性结合RNA以增加荧光强度，以及使用该试剂的白细胞的分类和计数方法，由此能够容易且高精度地分类和计数诸如未成熟白细胞和异常白细胞的异常细胞，同时 可以进行正常白细胞的时间，分类和计数以及白细胞的计数。
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公开(公告)号：US5891731A
公开(公告)日：1999-04-06
申请号：US843260
申请日：1997-04-14
申请人： Yasumasa Akai , Yuji Itose , Kayo Hatanaka , Takashi Sakata
发明人： Yasumasa Akai , Yuji Itose , Kayo Hatanaka , Takashi Sakata
CPC分类号： G01N33/5094 , G01N15/1459 , G01N2015/1006 , G01N2015/1402 , Y10S436/80 , Y10T436/10 , Y10T436/101666
摘要： A reagent for measuring reticulocytes which has at least one dye which specifically stains reticulocytes and at least one dye which specifically stains leukocytes.
摘要翻译： 用于测量网织红细胞的试剂，其具有至少一种特异性染色网织红细胞的染料和至少一种特异性染色白细胞的染料。
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公开(公告)号：US5308772A
公开(公告)日：1994-05-03
申请号：US903238
申请日：1992-06-23
申请人： Takashi Sakata , Mitsue Ito
发明人： Takashi Sakata , Mitsue Ito
CPC分类号： G01N33/5094 , G01N15/14 , Y10S436/80 , Y10T436/107497 , Y10T436/25
摘要： A hematological specimen for classifying and counting leukocytes with a flow cytometer is prepared. A sample to be assayed is prepared by eliminating influences of erythrocytes from a hematological sample without changing leukocytes morphologically; adjusting the pH value to a level suitable for staining; and staining the leukocytes with at least two dyes including Astrazon Yellow 3G capable of specifically staining at least basophils and immature granulocytes and Neutral Red capable of specifically staining at least eosinophils. Thus leukocytes contained in the hematological sample can be classified at least three or six groups including immature granulocytes by measuring a single specimen with a flow cytometer.
摘要翻译： 制备用流式细胞仪分类和计数白细胞的血液标本。 通过消除来自血液样品的红细胞的影响而不形态地改变白细胞来制备要测定的样品; 将pH值调节至适于染色的水平; 并用至少两种染料染色白细胞，包括能够特异性染色至少嗜碱性粒细胞和未成熟粒细胞的Astrazon Yellow 3G，以及能够特异性染色至少嗜酸性粒细胞的中性红。 因此，通过用流式细胞仪测量单个样本，血液学样品中含有的白细胞可以分为至少三组或六组，包括未成熟粒细胞。
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