CN101262950B - 流量计量分析器 - Google Patents
流量计量分析器 Download PDFInfo
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Abstract
一种具有流量计量的血液学分析器或细胞计芯子系统。其可以具有芯上或芯外流量计量和控制。所述系统可以具有局部的直接流量测量,以提供准确的单位体积计数。对于芯子的射流回路检验而言,可以存在很多种方案。例子可以包括与回路的零流量、接口、压力、流速、流体类型和质量、回流、干燥鉴定、温度暴露极限等相关的以及与芯子相关项目相关的检验。
Description
本发明要求2005年7月1日提交的美国临时专利申请No.60/696162的权益。在此引入2005年7月1日提交的美国临时专利申请No.60/696162以供参考。
技术领域
本发明涉及分析器,具体涉及分析器中的流量计量。更具体而言,本发明涉及血液学分析器和流式细胞器中的流量计量。
背景技术
本发明涉及的专利和申请可以包括:2002年5月7日颁布的名为“Fluid Driving System for Flow Cytometry”的美国专利No.6382228;2003年7月22日颁布的名为“Portable Flow Cytometry”的美国专利No.6597438;2005年11月29日颁布的名为“Optical Alignment DetectionSystem”的美国专利No.6970245;2003年4月15日颁布的名为“OpticalDetection System for Flow Cytometry”的美国专利No.6549275;1998年11月17日颁布的名为“Electrostatically Actuated Mesopump Having aPlurality of Elementary Cells”的美国专利No.5836750;2004年12月30日提交的名为“Optical Detection System with Polarizing Beamsplitter”的美国专利申请No.11/027134;2005年5月16日提交的名为“CytometerAnalysis Cartridge Optical Configuration”的美国专利申请No.10/908543;以及2005年4月25日提交的名为“Flow Control System of a Cartridge”的美国专利申请No.10/908014;在此引入所有的这些专利和申请以供参考。
发明内容
本发明涉及血液学分析器和细胞计的部分的直接流量监视和测量,从而提高诸如单位体积血液计数的某些项目的准确度。而且,还可以实现对分析器或细胞计的各种检验。
附图说明
图1是颗粒计数和尺寸测量系统的方框图;
图2示出了用于白血球的示范性血液分析芯的一部分;
图3示出了用于红血球的示范性血液分析芯的一部分;
图4示出了用于血红蛋白的示范性血液分析芯的一部分;
图5示出了在当前系统中采用的流量传感器的示意图;
图6是带有流量感测,以补偿血液学分析器的处理器输出的方框图;
图7是带有流量感测和闭合环路,以控制向血液学分析器的流动的方框图;
图8示出了针对封闭通道的零流量检验的设置;
图9示出了用于分析器和芯接口检验的设置;
图10示出了用于射流回路的压力和/或流速检验的设置;
图11示出了用于流体类型和质量检验的传感器设置;
图12示出了用于回流检验的设置;
图13示出了用于射流芯的干燥鉴定的设置;
图14示出了用于射流回路的温度暴露极限检验的设置;以及
图15示出了用于芯外流量感测的设置。
具体实施方式
本发明总体上涉及样本分析器,具体而言,涉及在病人的护理点,例如,在医生的办公室、家庭或其他实地使用的带有可拆卸和/或一次性芯子的分析器。通过提供带有所有必需的反应剂和/或流体的可拆卸和/或一次性芯子,可以在实验室环境以外凭借少量的专业培训或者不靠专业培训来可靠地使用所述样本分析器。例如,其可以有助于简化样本分析处理,降低成本以及医疗人员或其他人员的负担,并且提高针对很多病人的样本分析的便利性,这些病人包括那些需要相对频繁地进行血液监测/分析的病人。
流式细胞计量术是一种允许对颗粒悬浮样本中的颗粒进行快速有效的识别的方法。在这种方法中,可以通过流动通道输送血样中的通常为细胞的悬浮颗粒,在所述通道中采用一个或多个聚焦光束照射样本中的各个颗粒。可以通过一个或多个光探测器探测光束与流经所述流动通道的各个颗粒之间的相互作用。通常,可以将所述探测器设计为测量特定射束或发射波长上的光吸收或荧光发射,和/或探测特定散射角上的光散射。因而,可以将穿过所述流动通道的每一颗粒识别为一个或多个与其吸收、荧光、光散射或者其他光或电特性相关的特征。通过所述探测器测量的特性能够将每一颗粒映射到特征空间内,所述特征空间的轴是由探测器测量的光强或其他特性。在理想的方法中,样本中的不同颗粒映射到特征空间的不同的非重叠区域内,由此根据颗粒在特征空间内的映射对每一颗粒进行分析。这样的分析可以包括对颗粒进行计数、识别、量化(关于一种或多种物理特性)和/或分类。
一个示范性的例子可以是一种样本分析器,将所述样本分析器设置为具有接收所采集的样本的可拆卸芯,例如,所采集的样本可以是所采集的全血样本,一旦安装了所述可拆卸芯,并启动了分析器,所述分析器和芯子就可以自动处理样本,所述分析器可以为用户提供足够的信息进行临床决断。在某些例子中,分析器将显示或打印定量结果(例如,处于预定范围之内和/或之外的),因而不需要用户进行额外的计算或数据分析。
例如,可以采用所述样本分析器确定血样中的白血球的数量和/或类型。在一个示范性例子中,所述分析器包括外壳和可拆卸射流芯,其中,所述外壳适于接纳可拆卸射流芯。在某些情况下,所述可拆卸射流芯是一次性芯子。在一个示范性的例子中,所述可拆卸射流芯可以包括一种或多种试剂(例如,球化剂(sphering agent)、溶血剂、染色剂和/或稀释剂)、一个或多个分析通道、一个或多个流量传感器、一个或多个阀门和/或射流回路,所述射流回路适合对样本进行处理(例如,球化、溶解、染色或其他),并将受到处理的样本提供给芯子上的适当分析通道。为了支持所述卡片,所述外壳可以包括,例如,压力源、一个或多个光源、一个或多个光探测器、处理器和电源。所述压力源可以向可拆卸射流芯端口提供适当的压力,从而根据需要驱动流体通过射流回路。可以采用分析器的一个或多个光源询问可拆卸芯的至少一个选定分析通道内的准备样本,所述分析器的一个或多个光探测器可以探测穿过样本、被样本吸收和/或受到样本散射的光。可以将所述处理器连接至所述光源和探测器中的至少一些上,并且所述探测器可以确定所述样本的一个或多个参数。在一些例子中,可拆卸射流芯上的一个或多个分析通道可以包括一个或多个流式细胞计量通道。在一些示范性的例子中,可以向所述可拆卸射流芯提供全血样本,所述可拆卸芯可以适于执行血液分析。
图1是示范性样本分析器10和芯子14的透视图。示范性样本分析器10可以包括外壳12和可拆卸或一次性芯子14。示范性外壳12可以包括基座16、盖18和将基座16附着到盖18上的铰链20,但这不是必需的。在所述示范性例子中,基座16包括第一光源22a、第二光源22b和第三光源22c,连同相关光学部件和样本分析器的操作所需的电子部件。可以具有更多或更少的光源。根据应用场合的不同,每一光源可以是单个光源或多个光源。在某些情况下,外壳的总尺寸可以明显小于四分之一立方英尺。同样地,外壳的总重量可以明显小于一磅。
示范性的盖12可以包括压力源(例如,带有控制微阀的压力室)、第一光探测器24a、第二光探测器22b和第三光探测器22c,每一光探测器带有相关的光学部件和电子部件。可以存在更多或更少的探测器。根据应用场合的不同,每一光探测器也可以是单个光探测器或多个光探测器。根据应用场合的不同,还可以结合偏振器和/或滤光片。所述示范性可拆卸芯14可以适于经由样本采集器端口接收样本流体,在示范性例子中,所述端口带有刺血针32。在某些例子中,所述刺血针32可以是能缩回的和/或弹簧加载的。可以采用帽38在不使用可拆卸芯14时保护样本采集器端口和/或刺血针32。
在示范性例子中,所述可拆卸芯14可以对全血样本执行血液分析。可以采用刺血针32刺破用户的手指,以获得血液样本,可以通过毛细作用将血液吸到可拆卸芯14的涂覆了抗凝血剂的毛细管内。可以将所述可拆卸芯14构造为具有射流回路,某些射流回路是采用带有蚀刻通道的层压结构制造的。但是,可以设想通过任何适当的方式构造可拆卸芯14,包括注入模制或任何其他适当的制造工艺或方法。
在使用过程中,以及在将血样吸入到可拆卸芯14内之后,可以在盖18处于开启位置时将可拆卸芯插入到所述外壳内。在某些情况下,可拆卸芯14可以包括用于接收基座16内的配准销28a和28b的孔26a和26b,其有助于提供仪器的不部分之间的对准和连接。所述可拆卸芯14还可以包括第一透明流动流窗口30a、第二透明流动流窗口30b和第三透明窗口30c,所述窗口分别与第一、第二和第三光源22a、22b和22c以及第一、第二和第三光探测器24a、24b和24c对准。
在将所述盖移动到关闭位置,并对所述系统加压时,盖18可以经由压力提供端口36a、36b、36c和36d分别向示范性可拆卸芯14中的压力接收端口34a、34b、34c和34d提供受控压力。根据应用的不同,可以设想采用更多或更少的压力提供和压力接收端口。或者,或此外,可以设想在可拆卸芯14上设置一个或多个微泵,例如静电激励中泵(mesopump),以提供操作可拆卸芯14上的射流回路所需的压力。例如,在美国专利No5836750、6106245、6179586、6729856和6767190中描述了一些示范性静电激励中泵,在此引入所有的这些专利文献以供参考。一旦加压,所述的示范性的仪器就可以对所采集的血样执行血液分析。
当前系统可以提供基于微型流式细胞器或血液学分析器的全血细胞计数(CBC)卡,以获得一个或多个下述项目,包括:红血球(RBC)计数、球化RBC、血小板计数、RBC的溶解、白血球(WBC)的多部分差分计数、血红蛋白的基于吸收率的测量以及RBC、血小板、WBC、血红蛋白等的各种额外指标,当前系统还外加流体动力聚焦,以建立细胞的单行流,并提供气动流体驱动系统。额外的项目可以由当前系统提供和/或是当前系统的一部分。可以将细胞计和血液学分析器看作是相同或类似的系统。
图2是示出了芯子或卡14的WBC部分的示范性例子的一些方面的示意图。可以从将全血样本11采集到样本采集器13内入手。可以将血液推到位于浮动(fly)注入器33上的溶血剂(lyse)上。可以通过泵机构或流速控制盒35提供用于推动样本的流速以及溶解和包鞘流体。用于浮动注入器上的溶血剂的溶解流体可以来自溶血剂贮存器37。溶血剂流体和血液可以通过溶解通道39前进至流体动力聚焦室23。包鞘流体可以从鞘液贮存器25流到流体动力聚焦室23,以辅助白细胞穿过光通道29按照单行41对准,以供探测和分析。在细胞前进到光通道29之后,可以将细胞和流体运送到废料存储器31。
图3是示出了芯子或卡14的RBC部分的示范性例子的某些方面的示意图。这一卡14可以与WBC卡14类似,只是该卡是针对RBC分析设计的。类似地,可以将仪器10设计为用于RBC。可以从使全血样本11进入样本采集器13入手。可以将血液推导浮动注入器15上的球化剂上。可以通过泵机构或流速控制盒17提供用于推动样本的流速以及球化和包鞘流体。用于浮动注入器15上的球化剂的球化流体可以来自球化溶液贮存器19。所述溶液和血液可以通过所述球化通道21前进到流体动力聚焦室23。包鞘流体可以从鞘液贮存器25流到流体动力聚焦室23,以辅助球化红血球穿过光通道29按照单行27对准,以供探测和分析。在细胞穿过光通道29传输之后,可以将细胞和流体运送到废料存储器31。
图4是示出了芯子或卡14的血红蛋白(HGB)卡33或HGB部分的示范性例子的某些方面的示意图。该卡可以替代WBC卡14,只是该卡是针对HGB分析设计的。类似地,可以将仪器10设计为用于HGB测量。可以从将全血样本11采集到样本采集器13内入手。可以将血液推到吸收率测量试管43上。可以通过泵机构或流速控制盒45提供用于推动样本的流速。血液可以通过吸收率测量试管43传输,吸收率测量试管43可以提供吸收率测量47。在测量之后,可以将血液继续输送至废料贮存器31。
血液学分析器和流式细胞计(即分析器)可以采用处于开环中的注射器泵,并且其不对各种试剂和驱动流体的流速进行测量。在这样的分析器中采用流量传感器可以有助于提高分析器系统的总体准确度。对流的直接和局部测量可以为采用当前系统获得更加准确、精确的流速和每单位体积的血液计数提供基础。采用一次性分析卡的小型化血液学分析器可能需要流量传感器实现流速体积误差补偿。这对于开发提供某些测试的血液分析器是很关键的,所述测试在Clinical LaboratoryImprovement Amendments of 1988(CLIA)法规下可能被放弃。能够提供这样的被放弃的测试的分析器需要自诊断能力,从而针对气泡、流泄漏、闭塞等进行检验、探测和校正。
对小型化、低成本、超灵敏的稳定液体流量传感器的开发可以实现血液分析器中的流速和流体积(剂量)的测量。将流量传感器与闭环泵送系统结合使用能够实现对流速误差和变化的补偿。还可以将流量传感器用作测量仪表诊断传感器,可以通过将其设置在仪器或一次性卡的适当位置处来探测气泡、闭塞和流泄漏。这里是指用于本发明的能够测量非常低的流速的、具有小体积并且消耗很低的功率的流量传感器。
对于医疗、工业、生命科学和商业应用而言,对精确测量流体(即液体和气体)的流速的需求与日俱增。所预期的流速可以从针对给药、生命科学分析仪表检测和机器人液体处理系统的nL/min到针对透析设备和其他工业应用的mL/min乃至升/min。当前微流控技术申请可以采用必须通过某些传感器完成的某种形式的精确流体计量或流体定量配料,所述传感器具有超小尺寸(芯片尺寸小于25mm2),所耗功率低(小于75mW),具有高准确度(优于2%),响应速度快(快于1毫秒)。
可以采用直接或间接测量技术测量流速。通常通过测量静态属性,之后进行计算来间接测量流速。传统方法是测量填充已知体积所需的时间量。这种方法最大的优点是,可以以高精确度了解静态属性,并且可以将其追溯至国际标准。所述计算简单并且以经过验证的自然法则为基础,因而可以提高对结果的信心。这种方法存在弊端。一个明显的弊端在于无法了解流速随时间变化的程度。此外,测量越快,精确度越差。因而,这一技术最适合流速恒定的情况,以及响应时间/速度无关紧要的情况。
另一种间接流量测量方法可以利用有限空间内的流量与压降成正比的原理。可以采用差压传感器测量一定收缩下的压降,所述收缩包括由管道长度导致的限制。这种方法可以克服流速不稳定和长响应时间带来的限制(在时间体积法中),但是其代价是计算更为复杂,潜在的外部误差更大,并且介质的干扰更大。可以通过更小的限制获得更大的精确度,但是其直接改变流动特性。
人们对在实验室之外的实用基础上测量非常低的流量的兴趣越来越大。对于气体而言,其可以小于1升/分,对于液体而言,其可以小于1μL/分。在这样低的流量下,间接方法所固有的误差将被极大地放大,同时信号将被降低。为了避免这样的问题,可以在当前系统中提供对流速的直接测量。
液体流量传感器可以采用经验证的微机电系统(MEMS)技术提供非常小的封装尺寸内的快速、精确的流速测量,包括对非常小的速率的快速、精确测量。如图5所示,这样的流量传感器可以采用基于热的技术,其中,将隔热加热器(Rt)51加热至超过周围环境,并且其中,液体流速与位于加热器两侧的上游(Ru)温度传感器52和下游(Rd)温度传感器53之间的温度差成正比。针对传感器的校正曲线可以随着液体的热传导率而变化。
可以针对要求在1nL/min到50μL/min的极低流速下进行精确流量测量的应用对示例流量传感器进行特定设计。可以将这种传感器用于在这一区域内工作的反馈控制环。所述传感器可以以快速响应时间和自动温度补偿为特征。这一流量传感器可以采用基于MEMS的热测速技术测量经隔离的流动通道内的液体的质量流速。这样的传感器可以从新泽西州莫里森镇的Honeywell International公司获得。其他类似的流量传感器以及适当的压力传感器也可以从该公司获得。
图6是示例分析器系统60(例如,细胞计或血液学分析器)的示意图,所述分析器系统60具有射流回路61、连接至流量传感器65的泵压力源62和连接至流体回路/分析器66的废料贮存器64。射流回路61包括连接至流体回路/分析器66的流量传感器65。这一布局能够实现局部的直接流速测量。源62可以提供经由流量传感器65抵达流体回路/分析器66的流体。有关流体的信号67可以从分析器66到达处理器/控制器63。表示到分析器66的流体流的速度的信号68可以从流量传感器65传输至处理器63。可以将由从分析器66到处理器63的信号67得到的结果作为由流量传感器65感测的流速的函数进行校正。之后,处理器63可以输出校正结果69,所述结果可以包括血液计数,例如,每单位体积内的细胞的数量。
图7是示例分析器系统70(例如细胞计或血液学分析器)的示意图,所述分析器系统70具有与分析器系统60类似的部件,其同样允许局部的直接流速测量。但是,处理器63可以向泵/压力源62提供反馈信号71,由其得到了对流速传感器65经由信号68感测的由泵/压力源62提供的流速的闭环控制。之后,处理器63基于所述闭环控制提供结果72,其可以包括血液计数,例如,每单位体积的细胞的数量。
借助流量传感器的局部直接测量的当前系统60和70的目的在于获得精确的单位体积计数。例如,所述系统的光学部分每秒可以提供来自样本的若干个细胞。可以将这一数据除以(例如)以微升/秒为单位的当前流量传感器的输出,以获得每微升的细胞数。
图8示出了零流量检验的方法。可以以压力IN向通道81注入诸如液体的流体。在通道81内可以具有流量传感器82。沿管道再往下可以具有闭合的阀门83。借助阀门83,在向填充了液体的通道82的入口施加压强的同时流量传感器82探测的流速应当为零。如果传感器82这时读数不为零,那么在所述通道中可能存在气泡或泄漏。这一检验可以假设通道81的壁相对而言是非柔顺的,例如,是刚性的。
图9示出了针对分析器单元91和射流芯92之间的接口泄漏的方法。可以具有输出连接至流量传感器94的泵/压力源93,流量传感器94又连接至分析器93和芯子91之间的接口95。可以将流量传感器96连接于射流回路97和接口95之间。由流量传感器94探测的流速应当与流量传感器96探测的流速匹配,除非在接口95处存在泄漏。
图10示出了针对压力/流速检验的方法。通道101可以具有输入流102。可以将压力传感器103和流量传感器104放到通道101内。由压力传感器103探测的压力和流量传感器104探测的流速的比值应当处于预定范围内,除非在通道101内存在局部或完全的堵塞105、气泡106或其他异常结构。
图11示出了针对流体检验的方法。流动通道111可以具有处于适当位置的温度传感器112、热导率传感器113和粘滞度传感器114。流体的流115可以进入通道111。可以探测由传感器112、113、114和其他传感器指示的穿过流动通道111的流体的温度、热导率、粘滞度和其他特性,并且借助相关计算,流体类型及其特性应当与预期相同。如果不是,那么所述流体可能具有不正常的类型,可能具有吸收的湿气,其内可能具有细菌生长,可能受到了不适当的混合,可能具有沉淀出来的盐,可能由于储藏期限过期而变质,等等。
图12示出了针对回流检验的方法。在射流回路中可以存在通道121。可以将流量传感器122放到通道121内,其中,流123穿过所述通道。流量传感器122可以探测传感器122中的回流,所述回流在很多种情况下是不符合要求的。
图13示出了可以用来对射流芯131进行干燥鉴定的方法。可以存在气源132,其连接至端口133,经由通道或管道134连接至射流芯131,并且连接至背压传感器135。气源132可以将具有已知流速的气体,例如,氮气泵送到射流芯131的端口133内。可以判断由传感器135指示的测量背压是否处于指定的“良好”范围内。气源132可以将具有已知压强的气体,例如氮气泵送到射流芯131的端口133内,其中,采用流速传感器替代压力传感器135,可以判断所测得的流速是否处于指定的“良好”范围内。
图14示出了用于执行温度暴露极限检验的方法。在射流芯141回路内,可能存在连接至正常流动通道143以及连接至旁路通道144的输入通道142。可以在旁路通道144的入口放置由封闭蜡或其他适当的材料构成的温度熔断器145。可以采用这种方法判断射流芯141是否暴露于超过规定温度暴露极限的温度下。就高温极限检验而言,在芯子141暴露于超过高温极限的温度下时,温度熔断器145可以打开旁路通道145的入口(例如,通过石蜡的熔化)。旁路通道145的这一开启能够允许在将芯子141插入到分析器内时,差错能够被芯子分析器探测到。
就流温度极限而言,温度熔断器145可以涉及水或其他适当的材料,例如,在暴露于低于低温极限的温度下时发生收缩而无法返回至其原始尺寸的材料。在将芯子141暴露于这样的温度下时,熔断器145可能受到影响从而打开旁路通道145。旁路通道145的这一开启能够允许在将芯子141插入到分析器内时,差错能够被芯分析器探测到。
图15示出了针对芯外流量感测的方法。该图示出了连接至芯子分析器单元152的芯子151。可以存在处于芯子151和分析器单元152的端子之间的接口153、154和155。可以存在处于芯子151和分析器单元152的端子之间的额外的或备选的接口156和157。芯上流速探测可以采用芯外传感器。芯上流动通道158可以分别经由接口154和155从芯外路由至芯外流量传感器159,以进行流量测量。或者,或此外,可以将两个或更多芯外压力传感器161和162分别经由接口156和157流体连接至沿芯上流动通道158的两个点,以探测通道158内的流速。而且,芯外泵/压力源163可以经由芯外流量传感器164和接口153为芯上流动通道158提供流体流。
在本说明书中,某些内容可能具有假设或预言的性质,尽管这些内容可能是通过其他方式或语气陈述的。
尽管已经相对于至少一个示范性例子说明了本发明,但是在阅读了本说明书的基础上,很多变化和修改对于本领域技术人员而言将变得显而易见。因此,其目的在于在考虑现有技术的基础上对权利要求做出尽可能宽的解释,使之包括所有的此类变化和修改。
Claims (3)
1.一种微流体分析器系统,包括:
具有输入端口的射流回路;
连接至所述输入端口的流量传感器;
具有连接至所述流量传感器的输出的泵/压力源;以及
连接至所述流量传感器和所述射流回路的处理器;并且
其中:
将所述流量传感器处的流速发送至所述处理器;
将所述射流回路处的结果发送至所述处理器;
所述处理器根据所述流速执行结果校正;
所述射流回路具有血液学分析器;并且
所述流量传感器包括隔热加热器和位于隔热加热器两侧的上游温度传感器和下游温度传感器。
2.根据权利要求1所述的系统,其中:
所述流速为单位时间的体积;
所述射流回路处的结果是单位时间的计数;并且
来自所述处理器的结果是单位体积的计数。
3.一种微流体分析器系统,包括:
具有输入端口的射流回路;
连接至所述输入端口的流量传感器;
具有连接至所述流量传感器的输出的泵/压力源;以及
连接至所述流量传感器和所述射流回路的处理器;并且
其中:
所述处理器向所述泵/压力源提供信号;
所述处理器根据所述流量传感器执行对所述泵/压力源提供的流速的闭环控制;
所述流速为单位时间的体积;
所述射流回路处的结果是单位时间的计数;
来自所述处理器的结果是单位体积的计数;
所述射流回路具有血液学分析器;并且
所述流量传感器包括隔热加热器和位于隔热加热器两侧的上游温度传感器和下游温度传感器。
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2006
- 2006-06-30 EP EP06774517.4A patent/EP1901847B1/en active Active
- 2006-06-30 CN CN2006800313599A patent/CN101262950B/zh not_active Expired - Fee Related
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- 2006-06-30 US US11/428,289 patent/US8034296B2/en active Active
- 2006-06-30 JP JP2008519729A patent/JP5189976B2/ja active Active
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CN101262950A (zh) | 2008-09-10 |
JP5189976B2 (ja) | 2013-04-24 |
JP2008545141A (ja) | 2008-12-11 |
WO2007005973A3 (en) | 2007-06-14 |
CN101253401B (zh) | 2013-01-02 |
EP1901846B1 (en) | 2015-01-14 |
CN101252994B (zh) | 2011-04-13 |
CN101252994A (zh) | 2008-08-27 |
WO2007005973A2 (en) | 2007-01-11 |
EP1901847B1 (en) | 2015-04-08 |
EP1901846A2 (en) | 2008-03-26 |
CN101253401A (zh) | 2008-08-27 |
US8034296B2 (en) | 2011-10-11 |
US20070031289A1 (en) | 2007-02-08 |
EP1901847A2 (en) | 2008-03-26 |
JP2009500612A (ja) | 2009-01-08 |
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