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1. 发明授权

US07083936B2 Aminodextran compositions and conjugates and method of making and using them 失效
标题翻译：氨基葡聚糖组合物和缀合物及其制备和使用方法
公开(公告)号：US07083936B2
公开(公告)日：2006-08-01
申请号：US10633382
申请日：2003-08-01
申请人： Stephen L. Ledis , Olavi Siiman , Cynthia G. Healy
发明人： Stephen L. Ledis , Olavi Siiman , Cynthia G. Healy
IPC分类号： G01N33/567
CPC分类号： B82Y5/00 , A61K31/715 , A61K47/61 , A61K47/665 , A61K47/6883 , C08B37/0003 , C08B37/0021 , G01N33/548 , Y10S530/812 , Y10S530/813 , Y10S530/815 , Y10S530/816
摘要： A composition containing polydisperse aminodextran polymer molecules is soluble in an aqueous solution at a concentration of 10 mg/ml. The molecules therein have a narrow size distribution characterized by an average molecule mean hydrodynamic diameter of greater than 115 nm, a polydispersity index of between 0.10 and 0.47, an average molecular weight (MW) greater than 3 million daltons, and an amine content of greater than 50 amines per molecule. Similar soluble compositions contain the polymer molecules with an average MW of greater than 7 million daltons. These compositions are useful in forming reagents by conjugation with proteins for labeling cells. Methods of making these compositions and reagents from conventional mixtures of aminodextran polymers involve fractionation on column chromatography.
摘要翻译：含有多分散氨基糊精聚合物分子的组合物可溶于浓度为10mg / ml的水溶液中。其中的分子具有窄尺寸分布，其特征在于平均分子平均流体动力学直径大于115nm，多分散指数为0.10至0.47，平均分子量（MW）大于3百万道尔顿，胺含量较大每分子多于50个胺。类似的可溶性组合物含有平均MW大于700万道尔顿的聚合物分子。这些组合物可用于通过与用于标记细胞的蛋白质缀合形成试剂。从氨基葡聚糖聚合物的常规混合物制备这些组合物和试剂的方法包括在柱色谱上分级分离。

2. 发明授权

US5731206A Method and reagent system for isolation, identification and/or analysis of leukocytes from whole blood samples 失效
标题翻译：用于从全血样品中分离，鉴定和/或分析白细胞的方法和试剂系统
公开(公告)号：US5731206A
公开(公告)日：1998-03-24
申请号：US630868
申请日：1996-03-29
申请人： Stephen L. Ledis , Harold R. Crews , Timothy J. Fischer , Ted Sena
发明人： Stephen L. Ledis , Harold R. Crews , Timothy J. Fischer , Ted Sena
IPC分类号： G01N33/48 , G01N33/50 , G01N31/00
CPC分类号： G01N33/5094 , Y10T436/101666 , Y10T436/107497 , Y10T436/108331
摘要： A lytic reagent composition and a kit of a lytic reagent system are disclosed for the rapid isolation, identification and/or analysis of at least five subpopulations of leukocytes from a whole blood sample. The composition and system have application to any environment in which the study and/or analysis of the leukocyte fraction of whole blood requires their isolation in their native or near native state. The lytic reagent composition of this invention comprises saponin and a carboxylic acid of the formula RCOOH wherein R is H or a C.sub.1-3 aliphatic hydrocarbon radial optionally substituted by one or more carbonyl and/or hydroxy groups. The kit of a lytic reagent system comprises the lytic reagent composition and a quench reagent.
摘要翻译：公开了溶解试剂组合物和溶解试剂体系试剂盒，用于从全血样品中快速分离，鉴定和/或分析至少五个白细胞亚群。该组合物和系统适用于任何环境，其中全血白细胞分数的研究和/或分析需要其天然或接近天然状态的分离。本发明的溶解试剂组合物包含皂角苷和式RCOOH的羧酸，其中R是H或被一个或多个羰基和/或羟基取代的C 3-脂肪烃。裂解试剂系统的试剂盒包含裂解试剂组合物和淬灭试剂。

3. 发明授权

US5155044A Lysing reagent system for isolation, identification and/or analysis of leukocytes from whole blood samples 失效
标题翻译：用于从全血样品中分离，鉴定和/或分析白细胞的裂解试剂系统
公开(公告)号：US5155044A
公开(公告)日：1992-10-13
申请号：US808041
申请日：1991-12-12
申请人： Stephen L. Ledis , Harold R. Crews , Timothy J. Fischer , Ted Sena
发明人： Stephen L. Ledis , Harold R. Crews , Timothy J. Fischer , Ted Sena
IPC分类号： G01N33/50 , G01N33/569
CPC分类号： G01N33/5094 , G01N33/56972 , Y10T436/101666 , Y10T436/107497 , Y10T436/108331
摘要： A method and reagent system are disclosed for the rapid isolation, identification and/or analysis of leukocytes from a whole blood sample. The method and reagent system of this invention has application to any environment in which the study and/or analysis of the leukocyte fraction of whole blood requires their isolation in their native or near native state. One of the environments in which this invention can be used to advantage is in the performance of white cell differentiation on automated instrumentation designed for that purpose. In one of the preferred embodiments of this invention, the lytic reagent can contain a mixture of both formic and acetic acid, with the formic acid comprising the major component thereof and the acetic acid being present in only minor quantities, (if at all). This reagent system is used to selectively effect stromatolysis of red blood cells and create subtle modifications to the leukocyte population to enable their automated differentiation into five (5) sub-populations. The advantage and uniqueness of this reagent system is the surprising speed at which it is able to effect the foregoing objectives (generally less than 10 seconds at room temperature) and the ability to further differentiate the leukocyte population (notably, the granulocyte population). Following stromatolysis, a quenching agent is added to retard the reactivity of the lytic reagent system and, thus, inhibit any further dramatic changes to the leukocyte population. The treatment of the whole blood sample in the foregoing manner is further unique in that the leukocyte fraction of the sample has retained its characteristic immunochemical response.
摘要翻译：公开了用于从全血样品快速分离，鉴定和/或分析白细胞的方法和试剂系统。本发明的方法和试剂系统适用于其中全血白细胞分数的研究和/或分析需要在其天然或接近天然状态下分离的任何环境。本发明可用于有利的环境之一是在为此设计的自动化仪器上实现白细胞分化。在本发明的一个优选实施方案中，溶解剂可以含有甲酸和乙酸的混合物，其中甲酸包含其主要组分，乙酸仅以少量存在（如果有的话）。该试剂系统用于选择性地影响红细胞的间质分解，并对白细胞群体产生微妙的修饰，使其能够自动分化为五（5）个亚种群。该试剂系统的优点和独特之处在于其能够达到上述目的（通常在室温下通常小于10秒）以及进一步区分白细胞群体（特别是粒细胞群体）的能力的惊人速度。在层间裂解后，加入淬灭剂以延缓溶解试剂体系的反应性，从而抑制白细胞群体的任何进一步的剧烈变化。以上述方式对全血样品的处理是进一步的独特之处在于，样品的白细胞部分保持了其特有的免疫化学应答。

4. 发明授权

US4962038A Multi-purpose blood diluent and lysing agent for differential determination of lymphoid-myeloid population of leukocytes 失效
标题翻译：多用途血液稀释液和裂解剂用于差异测定白细胞淋巴样骨髓细胞群体
公开(公告)号：US4962038A
公开(公告)日：1990-10-09
申请号：US735245
申请日：1985-05-17
申请人： James H. Carter , Stephen L. Ledis , Harold R. Crews
发明人： James H. Carter , Stephen L. Ledis , Harold R. Crews
IPC分类号： G01N15/12 , G01N33/50 , G01N33/72
CPC分类号： G01N33/5094 , G01N15/1209 , G01N33/721 , Y10T436/10 , Y10T436/101666 , Y10T436/107497 , Y10T436/108331
摘要： An isotonic multipurpose blood diluent, and a method for use of this diluent with a weak lysing reagent system which is especially suitable for routine enumeration of traditional hemogram values, and also the determination of lymphoid-myeloid populations of leukocytes, particularly in automatic particle counting systems.This blood diluent is capable of affording accurate, reproducible test results. It is an osmotically balanced aqueous solution of preselected pH containing Procaine hydrochloride for maintaining erythrocyte morphology during operation, N-(2-acetamido)iminodiacetic acid (ADA) as a blood cell stabilizing agent, and bacteriostatic agents including sodium 1-hydroxypyridine-2-thione, and dimethylolurea which, together with the ADA, allow preferential determination of myeloid-lymphoid leukocytes, and other hematological values.The lysing agent is a mixture of an aqueous solution of at least one quaternary ammonium salt having surface active properties, and an alkali metal cyanide.
摘要翻译：一种等渗多用途血液稀释剂，以及一种使用该稀释液与弱溶解试剂系统的方法，该方法特别适用于常规计数传统的血红蛋白值，以及白细胞淋巴样骨髓细胞的测定，特别是在自动粒子计数系统。该血液稀释剂能够提供准确，可重复的测试结果。它是含有普鲁卡因盐酸盐的预选pH的渗透平衡水溶液，用于在手术期间保持红细胞形态，作为血细胞稳定剂的N-（2-乙酰氨基）亚氨基二乙酸（ADA））和抑菌剂包括1-羟基吡啶-2- 硫醇和二羟甲基脲，其与ADA一起允许优先测定骨髓淋巴样白血球和其他血液学值。裂解剂是具有表面活性的至少一种季铵盐的水溶液和碱金属氰化物的混合物。

5. 发明授权

US4521518A Multi-purpose blood diluent and lysing agent for differential determination of lymphoid-myeloid population of leukocytes 失效
公开(公告)号：US4521518A
公开(公告)日：1985-06-04
申请号：US395530
申请日：1982-07-06
申请人： James H. Carter , Stephen L. Ledis , Harold R. Crews
发明人： James H. Carter , Stephen L. Ledis , Harold R. Crews
IPC分类号： G01N15/12 , G01N33/50 , G01N33/48 , C09K3/00
CPC分类号： G01N15/1209 , G01N33/5094 , Y10T436/101666 , Y10T436/105831 , Y10T436/106664 , Y10T436/107497 , Y10T436/108331
摘要： An isotonic multipurpose blood diluent, and a method for use of this diluent with a weak lysing reagent system which is especially suitable for routine enumeration of traditional hemogram values, and also the determination of lymphoid-myeloid populations of leukocytes, particularly in automatic particle counting systems.This blood diluent is capable of affording accurate, reproducible test results. It is an osmotically balanced aqueous solution of preselected pH containing Procaine hydrochloride for maintaining erythrocyte morphology during operation, N-(2-acetamido)iminodiacetic acid (ADA) as a blood cell stabilizing agent, and bacteriostatic agents including sodium 1-hydroxypyridine-2-thione, and dimethylolurea which, together with the ADA, allow preferential determination of myeloid-lymphoid leukocytes, and other hematological values.The lysing agent is a mixture of an aqueous solution of at least one quaternary ammonium salt having surface active properties, and an alkali metal cyanide.

6. 发明授权

US4485175A Method for three-volume differential determination of lymphocyte, monocyte and granulocyte populations of leukocytes 失效
标题翻译：三体积微分测定白细胞淋巴细胞，单核细胞和粒细胞群体的方法
公开(公告)号：US4485175A
公开(公告)日：1984-11-27
申请号：US454926
申请日：1983-01-03
申请人： Stephen L. Ledis , Ted Sena , Harold R. Crews , James H. Carter, II
发明人： Stephen L. Ledis , Ted Sena , Harold R. Crews , James H. Carter, II
IPC分类号： G01N33/48 , G01N15/12 , G01N33/483 , G01N33/49 , G01N33/50 , G01N33/72
CPC分类号： G01N33/5094 , G01N15/1209 , Y10T436/101666 , Y10T436/107497
摘要： A method and reagent system is described for differential determination of three-populations of leukocytes (lymphocyte, monocyte and granulocyte), using an automatic counting system. The reagent system includes a blood diluent and lysing reagent. The blood diluent is an osmotically balanced aqueous solution of ingredients at a preselected pH for maintaining erythrocyte morphology, blood cell stabilizing, buffering and bacteriostatic action. The lysing reagent is a mixture of an aqueous solution of quaternary ammonium salts, which lysing reagent is added to the diluted blood under more mild conditions of concentration and at a slower rate than is current practice in order to obtain an unexpected volumetric modification of at least one of the three-populations of leukocytes, whereby volumetric differential analysis can be accomplished.
摘要翻译：描述了使用自动计数系统来区分白细胞（淋巴细胞，单核细胞和粒细胞）群体的方法和试剂系统。试剂系统包括血液稀释剂和裂解试剂。血液稀释剂是预选pH值的渗透平衡的成分水溶液，用于维持红细胞形态，血细胞稳定，缓冲和抑菌作用。裂解试剂是季铵盐水溶液的混合物，其在更温和的浓度条件下以较慢的速率将裂解试剂加入到稀释的血液中，目的是为了获得至少的意外的体积改性白细胞的三个群体之一，从而可以实现体积差异分析。

7. 发明授权

US5492833A Reticulocyte analyzing method and apparatus utilizing light scatter techniques 失效
标题翻译：利用光散射技术的网织红细胞分析方法和装置
公开(公告)号：US5492833A
公开(公告)日：1996-02-20
申请号：US306346
申请日：1994-09-15
申请人： Carlos M. Rodriguez , Stephen L. Ledis
发明人： Carlos M. Rodriguez , Stephen L. Ledis
IPC分类号： G01N33/48 , G01N15/10 , G01N15/14 , G01N33/49
CPC分类号： G01N15/1459 , G01N15/12 , G01N2015/1006 , G01N2015/1037 , G01N2021/4704 , G01N2021/4716 , G01N33/49 , Y10T436/25125
摘要： A reticulocyte analyzing method and apparatus in which a biological sample stream of ghosted red cells is passed into and through a point focused beam of light, such as laser light. A light detector structure is positioned with respect to the axis of the light beam to provide a light output pulse indicative of the passage of each cell. Electrically conductive contacts within the fluid stream can provide additional electrical pulse outputs of each cell. A staining reagent can be utilized with a ghosting reagent to further differentiate the reticulocytes. The light and the light and electronic produced output pulses or signals can be combined to define and quantify reticulocytes as distinct from other known cell types.
摘要翻译：一种网织红细胞分析方法和装置，其中将重影的红细胞的生物样品流传入并通过诸如激光的点聚焦光束。光检测器结构相对于光束的轴线定位，以提供指示每个单元通过的光输出脉冲。流体流内的导电触点可以提供每个电池的附加电脉冲输出。染色试剂可与重影试剂一起使用以进一步区分网织红细胞。光和光和电子产生的输出脉冲或信号可以组合以限定和定量网织红细胞，与其他已知的细胞类型不同。

8. 发明授权

US4358394A Process for preparing whole blood reference controls having long term stability 失效
标题翻译：制备具有长期稳定性的全血参照对照的方法
公开(公告)号：US4358394A
公开(公告)日：1982-11-09
申请号：US292091
申请日：1981-08-12
申请人： Harold R. Crews , David L. Chastain, Jr. , Stephen L. Ledis
发明人： Harold R. Crews , David L. Chastain, Jr. , Stephen L. Ledis
IPC分类号： G01N33/96 , G01N33/48 , C09K3/00
CPC分类号： G01N33/96 , G01N2496/05 , Y10T436/101666 , Y10T436/105831
摘要： A process for preparing whole blood reference controls having long term stability up to six months for devices using electronic means for whole blood determinations including platelet count and mean cell volume and red blood distribution width. The preconditioning diluent for the red blood cells consists essentially of an aqueous solution of lactose, sodium azide, and a non-ionic surfactant and is pH buffered and osmolality adjusted. The media of the whole blood control includes lactose, fungicides and antibiotics, and supplementary agents including purine nucleosides. It also includes additional components which alter red blood cell membranes including bile salts and cholic acid derivatives, phenothiazine compounds and the salts thereof having antihistamine properties, and 4-aminobenzoic acid ester derivatives and their salts having local anesthetic properties. In the process, the red blood cells are separated from the whole blood and preconditioned by the diluent to attain desired size distribution width and volume. The preconditioned red blood cells then are mixed in the media in suitable number for yielding the desired cell count for the whole blood reference control.
摘要翻译：一种用于制备全血参照对照的方法，对于使用电子方法进行全血测定的装置，包括血小板计数和平均细胞体积以及红血液分布宽度，对于长达6个月的长期稳定性。用于红细胞的预处理稀释剂基本上由乳糖，叠氮化钠和非离子表面活性剂的水溶液组成，并且是pH缓冲和重量摩尔渗透压浓度调节的。全血控制的媒体包括乳糖，杀真菌剂和抗生素，以及嘌呤核苷等补充剂。它还包括改变红细胞膜的其它组分，其包括胆汁盐和胆酸衍生物，吩噻嗪化合物及其具有抗组胺特性的盐，以及4-氨基苯甲酸酯衍生物及其具有局部麻醉性质的盐。在此过程中，将红细胞与全血分离并用稀释剂预处理，以获得所需的大小分布宽度和体积。然后将预处理的红细胞以适当的数量在培养基中混合，以产生用于全血参照对照的所需细胞计数。

9. 发明授权

US4264470A Selecting goat erythrocytes to simulate human platelets in hematologic reference controls 失效
标题翻译：选择山羊红细胞以在血液参照对照中模拟人血小板
公开(公告)号：US4264470A
公开(公告)日：1981-04-28
申请号：US36795
申请日：1979-05-07
申请人： David L. Chastain, Jr. , Harold R. Crews , Stephen L. Ledis
发明人： David L. Chastain, Jr. , Harold R. Crews , Stephen L. Ledis
IPC分类号： A61K35/14 , A61K38/16 , G01N33/49 , G01N33/96 , G01N33/48 , G01N35/00
CPC分类号： G01N33/96 , G01N2496/05 , Y10T436/101666 , Y10T436/106664 , Y10T436/107497 , Y10T436/108331
摘要： A method is described for preparing a human platelet analog for use as a reference control in particle analysis instrumentation of the Coulter type. Goat erythrocytes from individual goats which have been selected with respect to age, sex, selective breeding, a controlled environment, prior veterinary treatment, and diet are altered in size and/or combined with samples from several goats, as needed, to obtain a blend for use as a reference control for automated particle counting equipment in which the goat erythrocytes simulate in number, size and distribution the platelets in human whole blood. The controls are useful as a free-standing reference for determination of human platelets only, or can be added to whole blood reference controls for multi-parameter instruments.
摘要翻译：描述了一种制备人血小板类似物以用作库尔特型粒子分析仪器中的参考对照的方法。根据需要，根据年龄，性别，选择性育种，受控环境，先前的兽医治疗和饮食选择的来自单个山羊的山羊红细胞的大小和/或根据需要与来自几只山羊的样品组合以获得混合物用作自动粒子计数设备的参考对照，其中山羊红细胞模拟人全血中血小板的数量，大小和分布。这些对照作为仅用于确定人血小板的独立参考文献是有用的，或者可以添加到多参数仪器的全血参考对照中。

10. 发明授权

US5188935A Reagent system and method for identification, enumeration and examination of classes and subclasses of blood leukocytes 失效
标题翻译：用于鉴定，计数和检查血白细胞类和亚类的试剂系统和方法
公开(公告)号：US5188935A
公开(公告)日：1993-02-23
申请号：US611381
申请日：1990-11-13
申请人： Robert C. Leif , Stephen L. Ledis , Robert I. Feinberg
发明人： Robert C. Leif , Stephen L. Ledis , Robert I. Feinberg
IPC分类号： G01N33/50 , G01N33/569
CPC分类号： G01N33/50 , G01N33/56972 , Y10T436/107497
摘要： Specified classes and subclasses of leukocyte blood cells are identified by immunohematology procedures, based on utilization of antigenic determinants on the cell surface, their reactivity with antibodies which fluoresce under known circumstances, and identified by utilization of principles of flow cytometry or morphology.This invention particularly concerns improvements in the lysing and fixing method used prior to detection and identifying of the cells. In this method, a whole blood sample first is incubated with a reagent including antibodies to the cell subclass to be identified, the antibodies having directly or indirectly made fluorescently responsive to a particular light (e.g. argon ion laser). The red blood cells then are lysed with a reagent containing saponin. Next follows a leukocyte fixing treatment, preferably using a cross-linking dialdehyde composition, such as glyoxal or glutaraldehyde.Details are given as to time, temperature, concentration, and the use of additives and stains, as well as the use of particular monoclonal antibodies in the preferred procedure.
摘要翻译：基于细胞表面上的抗原决定簇的利用，其与已知情况下发荧光的抗体的反应性以及通过使用流式细胞术或形态学原理鉴定的免疫血液学方法来鉴定白细胞血细胞的指定类别和亚类。本发明特别涉及在检测和鉴定细胞之前使用的裂解和固定方法的改进。在该方法中，首先将全血样品与包括要鉴别的细胞亚类的抗体的试剂一起温育，所述抗体直接或间接地对特定的光进行荧光反应（例如氩离子激光）。然后用含有皂苷的试剂将红细胞裂解。接下来进行白细胞固定处理，优选使用交联二醛组合物，例如乙二醛或戊二醛。给出关于时间，温度，浓度以及添加剂和污渍的使用以及在优选方法中使用特定单克隆抗体的细节。

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