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    • 3. 发明授权
    • Method for classifying and counting leukocytes
    • 白细胞分类和计数方法
    • US06900023B1
    • 2005-05-31
    • US09388899
    • 1999-09-02
    • Berend HouwenFu-sheng WangHiroyuki FujimotoTakashi SakataYukio Hamaguchi
    • Berend HouwenFu-sheng WangHiroyuki FujimotoTakashi SakataYukio Hamaguchi
    • G01N33/49G01N15/14G01N33/50G01N33/53G01N33/536G01N33/569
    • G01N33/56972G01N33/5094G01N2333/70546Y10S435/962Y10S436/80Y10S436/824Y10S436/825Y10T436/101666Y10T436/107497Y10T436/13Y10T436/25Y10T436/25125
    • 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.
    • 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。