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    Indirect fluorescent assay of blood samples
    1.
    发明授权
    Indirect fluorescent assay of blood samples 失效
    血液样品的间接荧光检测

    公开(公告)号：US5460979A

    公开(公告)日：1995-10-24

    申请号：US192629

    申请日：1994-02-07

    申请人： Robert A. Levine ,  Stephen C. Wardlaw ,  Leon W. M. M. Terstappen ,  Thomas J. Mercolino ,  Diether J. Recktenwald

    发明人： Robert A. Levine ,  Stephen C. Wardlaw ,  Leon W. M. M. Terstappen ,  Thomas J. Mercolino ,  Diether J. Recktenwald

    IPC分类号： G01N33/543 ,  B01L3/14 ,  G01N33/49 ,  G01N33/537 ,  G01N33/569 ,  G01N33/58 ,  G01N33/538 ,  G01N33/546

    CPC分类号： B01L3/5021 ,  G01N33/491 ,  G01N33/5375 ,  G01N33/56972 ,  G01N33/585 ,  Y10S435/81 ,  Y10S435/967 ,  Y10S435/971 ,  Y10S435/973 ,  Y10S436/805 ,  Y10S436/81 ,  Y10S436/824 ,  Y10S436/829 ,  Y10T436/111666

    摘要： A patient's health is diagnosed by centrifuging blood samples in a transparent tube, which tube contains one or more groups of particles such as lyposomes or plastic beads of different densities for each group. Each group of density-defined particles carries antigens or antibodies which are specific to a complement antigen or antibody which may be in the blood sample being tested, and which are indicative of the patient's health. A label-tagged antibody which is specific to all bound antibody/antigen couples is added to the blood sample so as to form labelled antibody+antigen-antibody complexes (AAAC) in the blood sample. Upon centrifugation, the complexed particles will settle out in different areas in the tube according to the respective density of the particles, and the degree of label emission of the particle layers can enable qualitative or quantitative analyses of the blood sample to be made. Unbound labelled antibodies will be washed away from the complexed layers by the washing action of the descending blood cells during the centrifugation step. Unbound labelled antibodies will thus not interfere with the analysis.

    摘要翻译： 通过在透明管中离心血液样品来诊断患者的健康，该管含有一组或多组颗粒，例如每组的不同密度的溶血细胞或塑料珠。 每组密度定义的颗粒携带对可能在被测试的血液样品中的补体抗原或抗体特异性的抗原或抗体，并且其表示患者的健康。 向血液样品中加入对所有结合的抗体/抗原对特异性的标记标签抗体，以在血液样品中形成标记抗体+抗原 - 抗体复合物（AAAC）。 离心后，根据颗粒的相应密度，络合的颗粒将沉淀在管中的不同区域中，并且颗粒层的标签发射程度可以使得要进行血液样品的定性或定量分析。 在离心步骤中，未结合的标记抗体将通过下降血细胞的洗涤作用从复合层中洗去。 因此，未结合的标记抗体不会干扰分析。

    Apparatus for indirect fluorescent assay of blood samples
    2.
    发明授权
    Apparatus for indirect fluorescent assay of blood samples 失效
    血液样品间接荧光检测仪器

    公开(公告)号：US5342790A

    公开(公告)日：1994-08-30

    申请号：US969379

    申请日：1992-10-30

    申请人： Robert A. Levine ,  Stephen C. Wardlaw ,  Leon W. M. M. Terstappen ,  Thomas J. Mercolino ,  Diether J. Recktenwald

    发明人： Robert A. Levine ,  Stephen C. Wardlaw ,  Leon W. M. M. Terstappen ,  Thomas J. Mercolino ,  Diether J. Recktenwald

    IPC分类号： G01N33/543 ,  B01L3/14 ,  G01N33/49 ,  G01N33/537 ,  G01N33/569 ,  G01N33/58 ,  G01N33/536

    CPC分类号： B01L3/5021 ,  G01N33/491 ,  G01N33/5375 ,  G01N33/56972 ,  G01N33/585 ,  Y10S435/81 ,  Y10S435/967 ,  Y10S435/971 ,  Y10S435/973 ,  Y10S436/805 ,  Y10S436/81 ,  Y10S436/824 ,  Y10S436/829 ,  Y10T436/111666

    摘要： A patient's health is diagnosed by centrifuging blood samples in a transparent tube, which tube contains one or more groups of particles such as lyposomes or plastic beads of different densities for each group. Each group of density-defined particles carries antigens or antibodies which are specific to a complement antigen or antibody which may be in the blood sample being tested, and which are indicative of the patient's health. A label-tagged antibody which is specific to all bound antibody/antigen couples is added to the blood sample so as to form labeled antibody+antigen-antibody complexes (AAAC) in the blood sample. Upon centrifugation, the complexed particles will settle out in different areas in the tube according to the respective density of the particles, and the degree of label emission of the particle layers can enable qualitative or quantitative analyses of the blood sample to be made. Unbound labeled antibodies will be washed away from the complexed layers by the washing action of the descending blood cells during the centrifugation step. Unbound labeled antibodies will thus not interfere with the analysis.

    摘要翻译： 通过在透明管中离心血液样品来诊断患者的健康，该管含有一组或多组颗粒，例如每组的不同密度的溶血细胞或塑料珠。 每组密度定义的颗粒携带对可能在被测试的血液样品中的补体抗原或抗体特异性的抗原或抗体，并且其表示患者的健康。 向血液样品中加入对所有结合的抗体/抗原对特异性的标记标签抗体，以在血液样品中形成标记抗体+抗原 - 抗体复合物（AAAC）。 离心后，根据颗粒的相应密度，络合的颗粒将沉淀在管中的不同区域中，并且颗粒层的标签发射程度可以使得要进行血液样品的定性或定量分析。 在离心步骤中，未结合的标记抗体将通过下降血细胞的洗涤作用从复合层中洗去。 因此，未结合的标记抗体不会干扰分析。

    Assay of blood or other biologic samples for target analytes
    4.
    发明授权
    Assay of blood or other biologic samples for target analytes 失效
    测定目标分析物的血液或其他生物样品

    公开(公告)号：US5776710A

    公开(公告)日：1998-07-07

    申请号：US771507

    申请日：1996-12-23

    申请人： Robert A. Levine ,  Stephen C. Wardlaw ,  Leon W. M. M. Terstappen ,  Kristen L. Manion ,  Rodolfo R. Rodriguez ,  Adrien P. Malick ,  Subhash Dhanesar ,  Stephen J. Lovell ,  Alvydas J. Ozinskas

    发明人： Robert A. Levine ,  Stephen C. Wardlaw ,  Leon W. M. M. Terstappen ,  Kristen L. Manion ,  Rodolfo R. Rodriguez ,  Adrien P. Malick ,  Subhash Dhanesar ,  Stephen J. Lovell ,  Alvydas J. Ozinskas

    IPC分类号： G01N33/543 ,  B01L3/14 ,  G01N33/49 ,  G01N33/537 ,  G01N33/569 ,  G01N33/58 ,  G01N33/558

    CPC分类号： B01L3/5021 ,  G01N33/491 ,  G01N33/5375 ,  G01N33/56972 ,  G01N33/585 ,  Y10S435/81 ,  Y10S435/967 ,  Y10S435/971 ,  Y10S435/973 ,  Y10S436/805 ,  Y10S436/81 ,  Y10S436/824 ,  Y10S436/829 ,  Y10T436/111666

    摘要： A patient's health may be diagnosed by centrifuging blood samples in a transparent tube, which tube contains one or more bodies or groups of bodies such as floats, inserts, liposomes, or plastic beads of different densities. Each density-defined body carries analyte-capture binding materials such as antigens or antibodies, which are specific to an epitope, or other specific high affinity binding site on a target analyte which target analyte may be in the blood or other sample being tested; and the level of which analyte is indicative of the patient's health. At least one labeled binding material which is also specific to an epitope, or other specific high affinity binding site on the target analyte is added to the sample so as to form labeled binding material/analyte/body complexes in the sample. Upon centrifugation, the complexes will settle out in different areas in the tube according to the respective density of the body or bodies; and the degree of label emission of the complex layers can enable qualitative and/or quantitative analyses of the sample to be made. Unbound labeled binding materials will be separated from the complexed layers by the washing action of ascending or descending components of the sample during the centrifugation step. Unbound labeled binding material will thus not interfere with the analysis.

    摘要翻译： 可以通过将透明管中的血液样品离心来诊断患者的健康，该管包含一个或多个不同密度的浮体，插入物，脂质体或塑料珠的主体或组。 每个密度定义的身体携带分析物 - 捕获结合材料，例如抗原或抗体，其对靶分析物是特异性的，或靶分析物上的其它特异性高亲和力结合位点，其目标分析物可能在待测试的血液或其他样品中; 并且其分析物的水平表示患者的健康。 将至少一种对靶分析物上的表位或其他特异性高亲和力结合位点特异性的标记结合物质加入到样品中，以便在样品中形成标记的结合材料/分析物/身体复合物。 离心后，复合物将根据身体或身体的相应密度沉淀在管中的不同区域; 并且复合层的标签发射程度可以使得要进行样品的定性和/或定量分析。 通过在离心步骤期间样品的上升或下降组分的洗涤作用，未结合的标记结合材料将从复合层分离。 因此，未结合的标签结合材料不会影响分析。

    Analysis of circulating tumor cells, fragments, and debris
    5.
    发明授权
    Analysis of circulating tumor cells, fragments, and debris 有权
    分析循环肿瘤细胞，碎片和碎片

    公开(公告)号：US08329422B2

    公开(公告)日：2012-12-11

    申请号：US12917630

    申请日：2010-11-02

    申请人： Galla Chandra Rao ,  Christopher Larson ,  Madeline Repollet ,  Herman Rutner ,  Leon W. M. M. Terstappen ,  Shawn Mark O'Hara ,  Steven Gross

    发明人： Galla Chandra Rao ,  Christopher Larson ,  Madeline Repollet ,  Herman Rutner ,  Leon W. M. M. Terstappen ,  Shawn Mark O'Hara ,  Steven Gross

    IPC分类号： G01N33/574 ,  G01N33/553 ,  G01N1/18

    CPC分类号： G01N33/574 ,  G01N33/54326 ,  Y10S436/813 ,  Y10T436/101666 ,  Y10T436/25125 ,  Y10T436/25375

    摘要： The methods and reagents described in this invention are used to analyze circulating tumor cells, clusters, fragments, and debris. Analysis is performed with a number of platforms, including flow cytometry and the CELLSPOTTER® fluorescent microscopy imaging system. Analyzing damaged cells has shown to be important. However, there are two sources of damage: in vivo and in vitro. Damage in vivo occurs by apoptosis, necrosis, or immune response. Damage in vitro occurs during sample acquisition, handling, transport, processing, or analysis. It is therefore desirable to confine, reduce, eliminate, or at least qualify in vitro damage to prevent it from interfering in analysis. Described herein are methods to diagnose, monitor, and screen disease based on circulating rare cells, including malignancy as determined by CTC, clusters, fragments, and debris. Also provided are kits for assaying biological specimens using these methods.

    摘要翻译： 本发明描述的方法和试剂用于分析循环肿瘤细胞，簇，碎片和碎片。 使用许多平台进行分析，包括流式细胞术和CELLSPOTTER®荧光显微镜成像系统。 分析损伤的细胞已被证明是重要的。 然而，有两个损害来源：体内和体外。 体内损伤是通过细胞凋亡，坏死或免疫应答发生的。 在样品采集，处理，运输，加工或分析过程中发生体外损伤。 因此，期望限制，减少，消除或至少限定体外损伤，以防止其干扰分析。 本文描述了基于循环稀有细胞（包括由CTC，簇，碎片和碎片确定的恶性肿瘤）来诊断，监测和筛选疾病的方法。 还提供了使用这些方法测定生物样品的试剂盒。

    Method and apparatus for imaging target components in a biological sample using permanent magnets
    6.
    发明授权
    Method and apparatus for imaging target components in a biological sample using permanent magnets 有权
    用于使用永磁体对生物样品中的目标成分进行成像的方法和装置

    公开(公告)号：US07828968B2

    公开(公告)日：2010-11-09

    申请号：US12031807

    申请日：2008-02-15

    申请人： Arjan G. J. Tibbe ,  Leon W. M. M. Terstappen

    发明人： Arjan G. J. Tibbe ,  Leon W. M. M. Terstappen

    IPC分类号： B03C1/02 ,  B03C1/30

    CPC分类号： G01N33/582 ,  G01N33/54326 ,  G01N33/56972 ,  G01N2333/70514 ,  G01N2446/00

    摘要： A system for enumeration of cells in fluids by image cytometry is described for assessment of target populations such as leukocyte subsets in different bodily fluids or bacterial contamination in environmental samples, food products and bodily fluids. Briefly, fluorescently labeled target cells are linked to magnetic particles or beads. In one embodiment, a small, permanent magnet is inserted directly into the chamber containing the labeled cells. The magnets are coated with PDMS silicone rubber to provide a smooth and even surface which allows imaging on a single focal plane. The magnet is removed from the sample and illuminated with fluorescent light emitted by the target cells captured by a CCD camera. In another embodiment, a floater having a permanent magnet allows the target cells to line up along a single imaging plane within the sample solution. Image analysis can be performed with a novel algorithm to provide a count of the cells on the surface, reflecting the target cell concentration of the original sample.

    摘要翻译： 描述了通过图像细胞计数法检测流体细胞的系统，用于评估目标群体，例如不同体液中的白细胞亚群或环境样品，食品和体液中的细菌污染物。 简言之，荧光标记的靶细胞与磁性颗粒或珠粒连接。 在一个实施例中，将小的永磁体直接插入到包含标记电池的腔室中。 这些磁体用PDMS硅橡胶涂覆，以提供平滑和均匀的表面，允许在单个焦平面上进行成像。 从样品中除去磁体，并用由CCD照相机俘获的靶细胞发射的荧光照射。 在另一个实施例中，具有永磁体的浮子允许靶细胞沿着样品溶液内的单个成像平面排列。 可以用新颖的算法进行图像分析，以提供表面上的细胞计数，反映原始样品的靶细胞浓度。

    Affinity-binding separation and release of one or more selected subset of biological entities from a mixed population thereof
    7.
    发明授权
    Affinity-binding separation and release of one or more selected subset of biological entities from a mixed population thereof 失效
    从其混合群体中亲和力结合分离和释放一个或多个选定的生物实体子集

    公开(公告)号：US5646001A

    公开(公告)日：1997-07-08

    申请号：US395967

    申请日：1995-02-28

    申请人： Leon W. M. M. Terstappen ,  Galla C. Rao ,  Dhanesh I. Gohel ,  Brian P. Feeley ,  Steven Gross ,  Ellen S. Church ,  Paul A. Liberti

    发明人： Leon W. M. M. Terstappen ,  Galla C. Rao ,  Dhanesh I. Gohel ,  Brian P. Feeley ,  Steven Gross ,  Ellen S. Church ,  Paul A. Liberti

    IPC分类号： A23L3/32 ,  B03C1/035 ,  G01N33/543 ,  G01N33/569 ,  G01N35/00 ,  G01N33/567

    CPC分类号： A23L3/32 ,  B03C1/035 ,  G01N33/54326 ,  G01N33/569 ,  G01N35/0098 ,  Y10S435/962 ,  Y10S435/973 ,  Y10S436/824 ,  Y10S436/825 ,  Y10T436/25

    摘要： A method for separation of a mixture of biological entities into at least three distinct, subpopulations. Different antibodies are provided, with each antibody bound to a solid support in a unique manner such that by a manipulation of the physical or chemical environment, the bonds between the antibodies and the solid supports can be selectively broken. The mixed population of cells is incubated with the antibodies. The cells are magnetically separated from a test medium and collected in a monolayer upon a collection surface. Then by manipulation of the physicochemical environment, specific linkages can be broken and desired cell subpopulations released from the collection surface. This method has medically significant diagnostic and therapeutic applications, as entire cell types can be separated from non-malignant medically vital cell types. Cancer can be diagnosed, staged, and monitored. Genetic analysis from maternal blood, CVS, or amniocentesis samples is possible. Diseases such as AIDS, tuberculosis or hepatitis can be monitored. This invention also has utility in the fields of bone marrow transplantation, fetal cell research, in vitro fertilization, and gene therapy.

    摘要翻译： 将生物实体的混合物分离成至少三个不同的亚群体的方法。 提供了不同的抗体，每种抗体以独特的方式结合到固体支持物上，使得通过操纵物理或化学环境，可以选择性地破坏抗体和固体支持物之间的键。 将混合的细胞群与抗体一起孵育。 将细胞与测试介质磁分离并收集在收集表面上的单层中。 然后通过操纵物理化学环境，可以破坏特定的连接并且从收集表面释放所需的细胞亚群。 该方法具有医学上显着的诊断和治疗应用，因为整个细胞类型可以与非恶性的医学上重要的细胞类型分离。 可以诊断，分期和监测癌症。 从母体血液，CVS或羊膜穿刺样品的遗传分析是可能的。 可以监测艾滋病，结核病或肝炎等疾病。 本发明还可用于骨髓移植，胎儿细胞研究，体外受精和基因治疗领域。

    Automatic lineage assignment of acute leukemias by flow cytometry
    8.
    发明授权
    Automatic lineage assignment of acute leukemias by flow cytometry 失效
    通过流式细胞仪自动谱系分配急性白血病

    公开(公告)号：US5605805A

    公开(公告)日：1997-02-25

    申请号：US326153

    申请日：1994-10-19

    申请人： Ben J. H. Verwer ,  Leon W. M. M. Terstappen

    发明人： Ben J. H. Verwer ,  Leon W. M. M. Terstappen

    IPC分类号： G01N15/14 ,  G01N33/574 ,  G01N33/536 ,  G01N21/64 ,  G01N33/49

    CPC分类号： G01N33/57426 ,  G01N15/1456 ,  G01N2333/705 ,  Y10S435/968 ,  Y10S435/973 ,  Y10S436/80 ,  Y10S436/813

    摘要： A method for automatic lineage assignment of acute leukemias. Eight four-parameter list mode data files are acquired with a flow cytometer in the following sequence: 1. unstained; 2. isotype controls; 3. CD10 FITC, CD19 PE; 4. CD20 FITC, CD5 PE; 5. CD3 FITC, CD22 PE; 6. CD7 FITC, CD33 PE; 7. HLADR FITC, CD13 PE and 8. CD34 FITC, CD38 PE. First, data files 3-8 are clustered employing an algorithm based on nearest neighbors. The clusters are then associated across the data files to form cell populations, using the assumption of light scatter invariance across tubes for each population. The mean positions of each cell population are compared to a decision tree which identifies normal cell populations. To identify leukemic cell populations, the algorithm eliminates normal cell populations from the data space and the remaining populations are classified as B-lineage ALL, T-lineage ALL, AML, AUL, B-CLL or unknown.

    摘要翻译： 一种自动谱系分配急性白血病的方法。 用流式细胞仪按以下顺序获取八个四参数列表模式数据文件：1.未染色; 2.同种型对照 CD10 FITC，CD19 PE; CD20 FITC，CD5 PE; 5.CDI FITC，CD22 PE; CD7 FITC，CD33 PE; 7. HLADR FITC，CD13 PE和8.CD34 FITC，CD38 PE。 首先，使用基于最近邻的算法对数据文件3-8进行聚类。 然后，使用每个群体的管的光散射不变性的假设，通过数据文件将簇相关联以形成细胞群体。 将每个细胞群体的平均位置与鉴定正常细胞群体的决策树进行比较。 为了鉴定白血病细胞群体，该算法从数据空间中消除正常细胞群体，其余群体分为B系ALL，T系ALL，AML，AUL，B-CLL或未知。