CN102046274A - 用于从流体分离生物分子的装置 - Google Patents
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Abstract
一种用于从流体分离生物分子的装置,该装置包括设置有至少一个微通道(1)的微流体部件,该至少一个微通道(1)的各壁中的至少一个(2)支撑多个纳米管或纳米线(9)。该部件包括电连接到所述纳米管或纳米线(9)的至少一部分的至少一个电极(11),并且该装置包括用于在电极和流体之间施加电压的构件(10)。纳米管或纳米线被分成几个活性区域(14),该几个活性区域(14)中的纳米管或纳米线(9)具有不同的密度。
Description
技术领域
本发明涉及用于从流体分离生物分子的装置,该装置包括设置有至少一个微通道的微流体部件,该至少一个微通道的至少一个壁支撑多个纳米管或纳米线,所述部件包括电连接到纳米管或纳米线的至少一部分的至少一个电极,所述装置包括用于在电极和流体之间施加电压的构件。
背景技术
存在芯片上实验室型(lab-on-a-chip type)微系统,用于对小尺寸的化学或生物样品进行分析和/或操作。由于持续的小型化,微电子和毫微电子技术使得越来越多的功能能够被集成在单个的微流体部件中。这些功能传统地存在于预处理样品、过滤样品、分离样品、以及检测样品等。
近来的发展使得使用碳纳米管成为可能。于是,国际专利申请WO-A-2006/122697描述了一种微流体部件,如图1所示,该微流体部件包括至少一个使流体流动的通道。通道1优选为封闭的通道,即,该通道1包括流体的入口和出口并且由底壁2、彼此面对的两个相对侧壁3a和3b以及顶壁4来限定。底壁2与侧壁3a和3b制作在支撑体中,该支撑体优选为硅基板,顶壁4可以由盖形成,优选地由密封到基板的盖形成。壁2、3a和3b中的至少一个支撑多个纳米管9。在通道中制作水平的纳米管(平行于底壁2)使得能够获得呈现出增加的处理表面的流体部件。
国际专利申请WO 01/63273描述了包括微流体部件的装置(图2),该微流体部件设置有由底壁2与两个侧壁3a和3b限定的微通道。微通道在其底壁2上支撑多个碳纳米管9。微通道1的每个端部包括贮液器5a和5b,贮液器5a和5b设计用来接收包括带电分子6的流体。设置在微通道1的每个端部的贮液器5a和5b分别包括负端子7和正端子8,使得能够沿着微通道1产生矢量为E的电场。电场使得存在于负端子7的贮液器中的充负电的分子通过电泳向正端子8的贮液器的方向移动。于是,微通道的纳米管形成分子筛,纳米管的间距根据分子的类型来进行调节。这样的装置要求制作不同的筛密度。此外,在某些情况下,分子尤其是DNA分子可能缠绕纳米管,形成难以清理的陷阱。
专利申请US-2004/0173506描述了使用纳米纤维以形成隔膜并控制分子的传输。隔开两个纳米纤维的间距典型地为能够通过隔膜的分子的最大尺寸。
专利申请US2007/0090026描述了通过传统的微电子技术来制作二维筛结构,以改善生物分子分离的速度和分辨度。筛结构通过光刻和反应离子蚀刻(RIE)技术在硅基板中进行蚀刻来制作,从而能够以亚微米的精度获得可控制的形貌。扁平的筛结构包括宽度为1μm且深度为300nm的平行主通道,该平行主通道通过宽度为1μm且深度为55nm的横向通道而彼此连接。诸如DNA分子和蛋白质分子的分子可以经由连接相邻的第一主通道和第二主通道的横向通道而从第一主通道到达第二主通道。装置的表面可以充负电。由此,与强充负电的分子相比,弱充负电的分子能够以更大的可能性从一个主通道到达另一个主通道。
目前,在各种不同的研究中提出的分离生物分子的分离装置主要存在难以工业化的缺陷,因为它们的制作成本很高。事实上,它们确实需要非常昂贵的光刻步骤来制作尺寸与相关分子的尺寸相对应的孔或者通道。
发明内容
本发明的目的在于提供用于从流体分离生物分子的装置,该装置不具有现有技术的缺陷。
通过权利要求,更具体地通过将纳米管或纳米线分成几个活性区域并且该几个活性区域中的纳米管或纳米线具有不同密度的事实来实现上述目的。
根据一改进处,活性区域的纳米管或纳米线的密度从一个区域到下一个区域沿着流体的流动方向增加。
根据一改进处,每个区域连接到不同的电极,从而装置包括将不同电压施加到不同电极的构件。
附图说明
通过下面对本发明的具体实施例的描述,其他优点和特征将更加清楚明显,本发明的具体实施例仅出于非限制性示例的目的给出并表示在附图中,其中:
图1图解了根据现有技术的微流体部件的截面图。
图2图解了根据现有技术的用于通过电泳分离生物分子的装置的透视图。
图3图解了根据本发明的装置的俯视图。
图4图解了沿图3的A-A剖取的截面图。
图5图解了图3的装置的俯视图,其中装置的盖已经被移除。
图6至图8图解了根据本发明的装置的纳米管与带电粒子或者未带电粒子的相互作用。
图9和图10以俯视图图解了本发明实施例的各种变形,其中没有示出盖。
图11以俯视图图解了第二实施例,其中没有示出盖。
图12以俯视图图解了第二实施例的变形,其中没有示出盖。
具体实施方式
根据图3至图5所示的具体实施例,用于从流体分离生物分子的装置包括设置有至少一个微通道1的微流体部件,该微通道1由底壁2和彼此面对的两个侧壁3a和3b限定。微通道1优选为封闭的微通道(图4),并且由包括用于流体通路的入口12和出口13的顶壁4限定。
可以通过在装置的入口12和出口13之间施加例如压力差而使流体在分离装置中流动。该压力差可以例如通过采用注射器的推杆、蠕动泵或者其他本领域技术人员所熟知的手段来施加。图3至图5中所述的微通道为直线形,但是它也可以具有曲线、螺旋线、圆形等形式。
由此,微流体部件可以制作在基板中,在该基板中挖有微通道以形成底壁2与侧壁3a和3b。顶壁4可以通过保护盖优选气密密封的保护盖形成,以获得封闭的且完全密封的微通道1。基板可以例如由硅制成。该装置还包括用于在微流体部件的电极11和流体之间施加电压的构件10。电极11可以通过局部掺杂硅基板而形成在微通道的一部分上或者通过采用完全掺杂的基板而形成。
微通道壁2,3a和3b中的至少一个支撑形成为阵列的多个导电纳米管9或纳米线。纳米管9优选与支撑纳米管9的一个壁或各个壁垂直。微流体部件的电极11电连接到至少一部分纳米管9。如果电极11通过掺杂基板而形成,则所有的纳米管自动连接到电极。DC电压V优选可调节的DC电压V通过施加电压的构件10而施加在电极11和流体之间。在图4中所示的具体实施例中,电压源连接到形成电极11的基板,并通过流体出口13连接到流体。
纳米管9例如可以由碳制成。碳具有导电的优点。通过使纳米管的表面电势变化,纳米管的表面由此使得每个纳米管9的静电相互作用的幅度被调制。填充微通道的流体优选为电解质(包含正离子和负离子的水溶液),其他基于极性溶剂的流体也是可能的。当电势V施加在纳米管和填充微通道的流体之间时,根据电势V的符号,纳米管本身被反型离子云(cloud of counterion)围绕,由此产生电荷和局部电场的非均匀分布。在双静电层的条件下这些反型离子的分布是已知的(这里,围绕圆柱体)。电势在纳米管的表面处等于V并渐近地减小到流体的电势。等电势面具有以纳米管为中心的圆筒形几何形状。电势减小的特征长度称作德拜长度。德拜长度不取决于静电电势,而是取决于填充微通道的流体或缓冲溶液的离子浓度,该浓度通常也被称作缓冲溶液的“离子强度”。当隔开纳米管的距离约为德拜长度或更小时,纳米管阵列形成由等电势线和垂直于等电势线的电场线限定的静电屏障(electrostatic barrier)。从而,当带电粒子或者分子接近已经施加有电势即被静电充电的纳米管时,具有与纳米管的符号相同的电荷的粒子或分子倾向于被排斥。
使分子能够根据其电荷而被分离的现象是基于分子的流体动力学直径。流体动力学直径(也表示为Dh)对应于分子的尺寸(或直径)适当地加两倍的德拜长度(德拜长度表示为λD)。德拜长度对应于当分子带电时围绕分子的双电气层的厚度。德拜长度具体地对应于当分子带电且包含在流体中时局部地平衡分子电荷的反型离子云的厚度。它取决于包括分子的流体的条件,具体地,取决于所存在的电解质的类型和浓度以及温度。
通过纳米管阵列构成的屏障,更具体地通过两个相邻的纳米管所限定的通路(passage)来进行包含在流体中的分子的分离。纳米管屏障优选垂直于微通道中的流体的流动方向,纳米管可以由底壁2或者侧壁3a和3b来支撑。根据替代实施例,纳米管可以由形成盖的顶壁4来支撑。
形成屏障的纳米管优选占据微通道的整个部分(section),以在整个部分上形成纳米管的排列。
两个相邻的纳米管9限定的通路对应于实际距离dr。从而,如图7所示,直径小于dr的小的未带电分子PM可以从两个相邻的纳米管之间通过,这不同于被限制的大的未带电分子GM。
在纳米管和填充微通道的流体之间施加电压V使得在两个相邻的纳米管之间能够获得可控制的有效距离de,如图6和8所示。有效距离由下面的公式表示:
-当纳米管阵列和分子通过相同符号的电荷静电充电时,de=dr-2λD,其中dr对应于隔开两个相邻的纳米管的距离并且λD对应于德拜长度;
-在其他情况下,具体地当纳米管阵列和分子具有相反的符号或者当如上参考图7所述的纳米管阵列和分子未被充电时,de=dr。
因此,如图8所示,纳米管之间的有效距离de选择为仅使所需类型的分子通过,并且更具体地,该有效距离根据要分离的分子的流体动力学直径Dh来选择。由此,尺寸基本相似的分子可以根据它们的电荷来分离,如图8所示。弱带电的分子MFC(小的反型离子云15)可以通过纳米管9的屏障,而强带电的分子MCE(大的反型离子云15)不能通过该屏障。
出于示例的目的,如果两个相邻的纳米管之间的有效距离de小于分子的流体动力学直径,则带正电的纳米管阵列可以限制带正电的分子。另一方面,如果纳米管阵列和分子通过相反符号的电荷充电,则分子的流体动力学直径比隔开纳米管阵列的两个相邻纳米管的实际距离dr大就足够了。
为了以可控制的方式使纳米管静电充电,用于施加电压的构件10使所施加的电压能够被调节,从而使得分子通过的可能性增大或减小。图6示出了具有不同电势17的两行纳米管9。对于直径相同的纳米管,碳纳米管9的静电相互作用的范围可以调制。
包含在流体中的分子的静电电荷还取决于构成流体的溶液的pH值。由此,可以根据分子所需的电荷来调节溶液的pH值,从而也能使分子的通路增大或减小。相关的分子通常是在特定pH范围内形成弱的负离子化酸的核酸或蛋白质(氨基酸的聚集体)。用作包含这些分子的缓冲溶液的流体于是可以是或多或少地带有盐的溶液。然后,带电分子本身被反型离子云围绕,取决于盐的浓度和成分该反型离子云的直径可以在几纳米到几十纳米的范围内。
连接到电极11的导电纳米管9的使用使得能够主动(实时)控制纳米管9的电势。这些纳米管9隔开几纳米,可以设想10nm的距离。隔开两个相邻纳米管的距离优选包括在1nm到20nm之间。从而,当电压施加到纳米管上时,它们可以形成具有与纳米管相同符号的电荷的带电分子的静电屏障。通过调节流体和纳米管之间的电压,可以调节纳米管的电势,由此调节静电屏障的透过性。
这样的装置可以根据纳米管之间的距离而用作筛并且/或者能够使不同电荷的分子被保持或被允许通过。
这样的装置由此可以用作分子的过滤和分离系统,但是它也可以用作使分子聚集的系统。在后面的情况下,关注的分子仅需要被保持在形成静电屏障的纳米管的部分的前面,而于此同时洗提(elute)更小的分子。然后,一旦位于屏障前面的保持区域富集关注的分子,则断开施加到屏障的电压,使得关注的分子能够通过并且能够收集到高度富集关注的分子的洗出液(eluate)。
前面描述的微流体部件的制造可以采用专利申请WO-A-2006/122697中描述的方法,并且可以采用电阻率优选为0.01Ω·cm的掺杂的硅基板。
根据图9所示的替代实施例,纳米管9分成为纳米管密度相似的几个活性区域14。每个活性区域14的纳米管9电连接到不同的对应电极(未示出)。该装置由此包括用于将不同电压施加到微流体部件的不同电极的构件。这样的对每个电极具有不同电寻址的装置使得能够在每个活性区域14的纳米管的级别(level)获得不同的电势。隔开两个相邻纳米管的有效距离de由此可以在每个活性区域14中不同地且实时地调节。在优选实施例中,该有效距离de可以从第一个区域到最后一个区域逐渐地减小,从而实现从一个区域到另一个区域的分子的逐渐分离。由此,可以通过单个装置隔离不同类型的分子。
根据图10所示的另一个替代实施例,不同活性区域14中的纳米管密度不同。例如,按照包含在流体中的分子的流动方向(在图10中从左到右),纳米管的密度逐区域地增加。导电纳米管可以都连接到单个电极(未示出),而该单个电极连接到用于在电极和流体之间施加电压的构件。从而,各纳米管具有相同的电势,并且包含在流体中的生物分子的分离根据流体流经的活性区域14的纳米管密度和电势而逐渐地进行。
在图10所示的替代实施例中,每个活性区域的纳米管电连接到不同的电极。该装置于是包括将不同的可变化的电压施加到每个电极的装置。这样的对每个电极具有不同电寻址的装置使得能够在每个活性区域的纳米管的级别获得不同的电势,从而实现从一个区域到另一个区域的分子的可实时调节的逐渐分离。
因此,纳米管密度的变化和/或施加在纳米管与流体之间的电压的变化可以用于限定将两个相邻的纳米管隔开的有效距离,并且相应地各分子的尺寸和/或电荷将易于通过这些纳米管形成的屏障或者被该屏障限制。
为了制造包括具有几个活性区域的微通道的装置,通过采用局部掺杂的硅基板以形成不同的电极,然后每个电极形成为其上形成有纳米管的活性区域14,国际专利申请WO-A-2006/122697中描述的方法被修改。
根据图11所示的另一个替代实施例,纳米管9可以具有行R的形式(位于图11的点线限定的区域中),以隔开两个相邻的毫微通道并形成相对于流体流动方向稍微倾斜地布置的屏障。隔开两行纳米管9的距离大于隔开同一行中的两个相邻纳米管的距离。在图11和12所示的具体实施例中,流体流动的大致方向控制为使纳米管的行相对于该方向倾斜地设置。在图11和12中,例如,流体入口12和出口13分别位于底部的左侧部和顶部的右侧部并且流体在压力的作用下设置在入口12和出口13之间,即相对于纳米管的行R倾斜地设置。只有流体动力学直径比隔开同一行的纳米管的有效距离de小的分子MFC能够通过由这些纳米管及它们的电势17形成的屏障并从而进入到图11和12的顶部毫微通道中。因此,在图11中,流体动力学直径比有效距离大的分子MCE被限制在底部毫微通道中,而分子MFC能够进入到顶部毫微通道中。由此,各分子可以根据它们的尺寸和/或电荷而被分类,从而两个相邻的毫微通道在它们靠近出口13的端部处包括不同尺寸和/或电荷的分子。
根据图12所示的替代实施例,不同的电势17,优选从底部到顶部增加,施加到纳米管9的每个行R,由此使得能够分离不同尺寸和/或电荷的分子。因此,在图12中,流体动力学直径比底部行的有效距离小而比纳米管的顶部行的有效距离de大的分子MCE被限制在中间的毫微通道中,分子MFC可以进入到顶部毫微通道中,并且流体动力学直径比纳米管的底部行的有效距离大的其他分子保留在底部毫微通道中。各行还可以具有不同的纳米管间的间隔,以对分子的尺寸和电荷的两个因素起作用。
用于施加电压的构件10可以包括浸在流体中的铂引线16(图4),或者本领域技术人员会采用的任何其他构件。
上述实施例能够使任意组成的混合物的分子被分离,该任意组成的混合物例如为核酸的混合物和/或蛋白质的混合物和/或缩氨酸的混合物。通过实时调节施加到纳米管的电压,上述分离可以连续地进行。
此外,在流体和纳米管之间施加电压使得装置的清洁更容易,尤其是当DNA分子缠绕在纳米管周围时,在纳米管上施加电势使得能够移除缠绕的分子。
上述装置可以包括能够并行处理分子的多个微通道。
本发明并不限于前面所述的各实施例,具体地,纳米管可以由导电纳米线优选地掺杂的硅制成的导电纳米线替代。
Claims (7)
1.一种用于从流体分离生物分子的装置,该装置包括设置有至少一个微通道(1)的微流体部件,该至少一个微通道(1)的各壁(2,3a,3b)中的至少一个支撑多个纳米管或纳米线(9),所述部件包括电连接到所述纳米管或纳米线(9)的至少一部分的至少一个电极(11),并且所述装置包括用于在该电极和该流体之间施加电压的构件(10),所述装置的特征在于所述纳米管或纳米线被分成几个活性区域(14),该几个活性区域(14)中的纳米管或纳米线(9)具有不同的密度。
2.根据权利要求1所述的装置,其特征在于每个区域(14)的密度从一个区域到下一个区域沿着所述流体的流动方向增加。
3.根据权利要求1至2中任一项所述的装置,其特征在于每个区域(14)连接到不同的电极,并且所述装置包括用于将不同电压施加到所述不同的电极的构件(10)。
4.根据权利要求1至3中任一项所述的装置,其特征在于所述纳米管或纳米线(9)在所述微通道中形成与所述流体的流动方向垂直的屏障。
5.根据权利要求1至3中任一项所述的装置,其特征在于所述纳米管或纳米线(9)在所述微通道中形成相对于所述流体的流动方向倾斜的屏障。
6.根据权利要求1至5中任一项所述的装置,其特征在于所述纳米管(9)由碳制成。
7.根据权利要求1至5中任一项所述的装置,其特征在于所述纳米线(9)由掺杂的硅制成。
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CN103890567A (zh) * | 2011-10-27 | 2014-06-25 | 惠普发展公司,有限责任合伙企业 | 用于过滤物种的装置 |
US9638625B2 (en) | 2011-10-27 | 2017-05-02 | Hewlett-Packard Development Company, L.P. | Apparatus for filtering species |
CN108348827A (zh) * | 2015-10-23 | 2018-07-31 | 惠普发展公司,有限责任合伙企业 | 纳米线微孔结构 |
US11213791B2 (en) | 2015-10-23 | 2022-01-04 | Hewlett-Packard Development Company, L.P. | Nano wire microporous structure |
CN108348827B (zh) * | 2015-10-23 | 2022-04-08 | 惠普发展公司,有限责任合伙企业 | 纳米线微孔结构 |
CN110462367A (zh) * | 2017-02-17 | 2019-11-15 | 斯泰特皮尔股份公司 | 过滤装置 |
CN113546698A (zh) * | 2020-04-24 | 2021-10-26 | 京东方科技集团股份有限公司 | 微纳流控芯片及其制造方法、微纳流控系统 |
CN113546698B (zh) * | 2020-04-24 | 2022-08-23 | 京东方科技集团股份有限公司 | 微纳流控芯片及其制造方法、微纳流控系统 |
CN112295724A (zh) * | 2020-09-14 | 2021-02-02 | 杭州电子科技大学 | 一种不同再生程度粉末活性炭浮选方法及浮选装置 |
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EP2282827A2 (fr) | 2011-02-16 |
JP2011520117A (ja) | 2011-07-14 |
FR2930900A1 (fr) | 2009-11-13 |
US20110108424A1 (en) | 2011-05-12 |
WO2009141528A3 (fr) | 2010-01-21 |
FR2930900B1 (fr) | 2010-09-10 |
CN102046274B (zh) | 2014-07-30 |
WO2009141528A2 (fr) | 2009-11-26 |
EP2282827B1 (fr) | 2013-07-31 |
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