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CN111479670B - 在样品处理装置中制作易碎密封件的方法 - Google Patents

在样品处理装置中制作易碎密封件的方法 Download PDF

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Publication number
CN111479670B
CN111479670B CN201880080923.9A CN201880080923A CN111479670B CN 111479670 B CN111479670 B CN 111479670B CN 201880080923 A CN201880080923 A CN 201880080923A CN 111479670 B CN111479670 B CN 111479670B
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China
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reverse
die
sealing
substrate
sample processing
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CN201880080923.9A
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CN111479670A (zh
Inventor
A·C·丹尼森
C·C·斯宾塞
J·P·林奇
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F Hoffmann La Roche AG
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F Hoffmann La Roche AG
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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B29—WORKING OF PLASTICS; WORKING OF SUBSTANCES IN A PLASTIC STATE IN GENERAL
    • B29C—SHAPING OR JOINING OF PLASTICS; SHAPING OF MATERIAL IN A PLASTIC STATE, NOT OTHERWISE PROVIDED FOR; AFTER-TREATMENT OF THE SHAPED PRODUCTS, e.g. REPAIRING
    • B29C66/00—General aspects of processes or apparatus for joining preformed parts
    • B29C66/01—General aspects dealing with the joint area or with the area to be joined
    • B29C66/05—Particular design of joint configurations
    • B29C66/10—Particular design of joint configurations particular design of the joint cross-sections
    • B29C66/11—Joint cross-sections comprising a single joint-segment, i.e. one of the parts to be joined comprising a single joint-segment in the joint cross-section
    • B29C66/112—Single lapped joints
    • B29C66/1122—Single lap to lap joints, i.e. overlap joints
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B29—WORKING OF PLASTICS; WORKING OF SUBSTANCES IN A PLASTIC STATE IN GENERAL
    • B29C—SHAPING OR JOINING OF PLASTICS; SHAPING OF MATERIAL IN A PLASTIC STATE, NOT OTHERWISE PROVIDED FOR; AFTER-TREATMENT OF THE SHAPED PRODUCTS, e.g. REPAIRING
    • B29C66/00—General aspects of processes or apparatus for joining preformed parts
    • B29C66/80—General aspects of machine operations or constructions and parts thereof
    • B29C66/83—General aspects of machine operations or constructions and parts thereof characterised by the movement of the joining or pressing tools
    • B29C66/832—Reciprocating joining or pressing tools
    • B29C66/8322—Joining or pressing tools reciprocating along one axis
    • B29C66/83221—Joining or pressing tools reciprocating along one axis cooperating reciprocating tools, each tool reciprocating along one axis
    • B—PERFORMING OPERATIONS; TRANSPORTING
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    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B01L3/502707—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by the manufacture of the container or its components
    • B—PERFORMING OPERATIONS; TRANSPORTING
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    • B29C66/43—Joining a relatively small portion of the surface of said articles
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    • B29C66/7392—General aspects of processes or apparatus for joining preformed parts characterised by the composition, physical properties or the structure of the material of the parts to be joined; Joining with non-plastics material characterised by the intensive physical properties of the material of the parts to be joined, by the optical properties of the material of the parts to be joined, by the extensive physical properties of the parts to be joined, by the state of the material of the parts to be joined or by the material of the parts to be joined being a thermoplastic or a thermoset characterised by the material of the parts to be joined being a thermoplastic or a thermoset characterised by the material of at least one of the parts being a thermoplastic
    • B29C66/73921—General aspects of processes or apparatus for joining preformed parts characterised by the composition, physical properties or the structure of the material of the parts to be joined; Joining with non-plastics material characterised by the intensive physical properties of the material of the parts to be joined, by the optical properties of the material of the parts to be joined, by the extensive physical properties of the parts to be joined, by the state of the material of the parts to be joined or by the material of the parts to be joined being a thermoplastic or a thermoset characterised by the material of the parts to be joined being a thermoplastic or a thermoset characterised by the material of at least one of the parts being a thermoplastic characterised by the materials of both parts being thermoplastics
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B29—WORKING OF PLASTICS; WORKING OF SUBSTANCES IN A PLASTIC STATE IN GENERAL
    • B29C—SHAPING OR JOINING OF PLASTICS; SHAPING OF MATERIAL IN A PLASTIC STATE, NOT OTHERWISE PROVIDED FOR; AFTER-TREATMENT OF THE SHAPED PRODUCTS, e.g. REPAIRING
    • B29C66/00—General aspects of processes or apparatus for joining preformed parts
    • B29C66/80—General aspects of machine operations or constructions and parts thereof
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    • B29C66/814—General aspects of the pressing elements, i.e. the elements applying pressure on the parts to be joined in the area to be joined, e.g. the welding jaws or clamps characterised by the design of the pressing elements, e.g. of the welding jaws or clamps
    • B29C66/8141—General aspects of the pressing elements, i.e. the elements applying pressure on the parts to be joined in the area to be joined, e.g. the welding jaws or clamps characterised by the design of the pressing elements, e.g. of the welding jaws or clamps characterised by the surface geometry of the part of the pressing elements, e.g. welding jaws or clamps, coming into contact with the parts to be joined
    • B29C66/81427—General aspects of the pressing elements, i.e. the elements applying pressure on the parts to be joined in the area to be joined, e.g. the welding jaws or clamps characterised by the design of the pressing elements, e.g. of the welding jaws or clamps characterised by the surface geometry of the part of the pressing elements, e.g. welding jaws or clamps, coming into contact with the parts to be joined comprising a single ridge, e.g. for making a weakening line; comprising a single tooth
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B29—WORKING OF PLASTICS; WORKING OF SUBSTANCES IN A PLASTIC STATE IN GENERAL
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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B29—WORKING OF PLASTICS; WORKING OF SUBSTANCES IN A PLASTIC STATE IN GENERAL
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    • B29—WORKING OF PLASTICS; WORKING OF SUBSTANCES IN A PLASTIC STATE IN GENERAL
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Abstract

描述了一种密封装置，所述密封装置被配置用于在样品处理装置中产生易碎的密封件。还描述了使用所述装置在样品处理装置中创建一个或多个易碎密封件的方法。

Description

在样品处理装置中制作易碎密封件的方法
背景技术
临床样品或环境样品中核酸的分析通常涉及对流体样品进行一系列化学、光学、电气、机械和热处理步骤。近年来，对开发用于进行生物样品分析以用于各种诊断和监测目的的一次性药筒的兴趣日益增长。例如，Wilding的美国专利5,587,128公开了通过进行多核苷酸扩增反应来用于扩增样品中预选的多核苷酸的装置。Anderson等人的美国专利5,922,591描述了一种小型的、集成的核酸诊断装置和系统。设备通常能够结合一种或多种样品分析操作来执行一种或多种样品获取操作和制备操作。
由于必须使用完全集成的自含式装置来执行一系列复杂的操作，因此制造商已经生产出这种设备，其中包含用于执行预装在设备中的测定所需的每个操作所必需的所有试剂。因此，可用测定的菜单非常有限，并且需要使用户能够并入用户提供的试剂，例如引物和探针，使得可以扩展可用的测定菜单以满足客户广泛的需求。
发明内容
本公开提供了一种密封压机，所述密封压机包括一对等温加热模具，所述一对加热模具包括正向模具和反向模具，所述正向模具和所述反向模具各自包括彼此相对且适于将沿着密封平面定位的样品处理装置保持在其间的密封表面，其中，所述正向模具和所述反向模具各自独立地定位在正向对准滑块和反向对准滑块上并且分别由其控制，所述正向对准滑块和所述反向对准滑块连接到致动器，所述致动器提供用于使所述正向模具和所述反向模具同步运动的推力。在一些实施方案中，正向对准滑块和反向对准滑块各自安装到正向卡钳臂和反向卡钳臂，并且正向卡钳臂和反向卡钳臂分别安装到连接到致动器的机械联动装置。在某些实施方案中，机械联动装置包括杠杆、第一连杆和第二连杆，其中，杠杆包括中心枢轴点以及第一末端和第二末端，其中，第一连杆枢转地连接到杠杆的第一末端并且枢转地连接到反向卡钳臂的一个末端，其中，第二连杆枢转地连接到杠杆的第二末端并且枢转地连接到正向卡钳臂的一个末端，其中，第一卡钳臂和第二卡钳臂分别连接到正向对准滑块和反向对准滑块。在其他实施方案中，密封压机进一步包括独立的正向模具控制机构，所述机构连接到正向模具并且包括连接到补充的正向模具线性滑块和独立的正向模具控制器的正向模具驱动器，其中，所述机构通过正向卡钳臂独立地致动正向模具，从而调节密封表面之间的接触压力。在一些实施方案中，一对等温加热模具被配置用于被加热到低于样品处理装置的基底的熔融温度的恒定温度。在某些实施方案中，基底包括聚丙烯，并且将模具加热至约105℃至115℃。
还提供了一种在包括管基底的样品处理装置中产生易碎密封件的方法，方法包括：(i)将密封压机的密封模具保持在恒定温度下，例如，比管基底的熔融温度低约15℃至25℃之间；(ii)沿着密封压机的密封平面放置管基底；(iii)在小于或等于管基底的压缩屈服强度的压力下将一对模具的每个密封表面压入管基底中，持续足够的时间，以将热量通过管基底的外膜传递至管基底的内结合表面；以及(iv)撤回加热模具并保持部件无张力，以允许由步骤(iii)产生的部分熔融的密封件冷却且收缩。在一些实施方案中，提供了一种在样品处理装置中产生易碎密封件的方法，所述样品处理装置包括具有外表面和内表面的中空管基底，其中，方法是由包括一对等温加热模具的密封压机来执行的，所述一对模具包括正向模具和反向模具，各自包括彼此相对且适于将沿着密封平面定位的样品处理装置固持在其间的密封表面，其中，正向模具和反向模具各自独立地位于正向对准滑块和反向对准滑块上并且分别由其控制，正向对准滑块和反向对准滑块连接到致动器，所述致动器提供用于使正向模具和反向模具同步运动的推力，方法包括：(a)将一对等温加热模具保持在低于样品处理装置的基底的熔融温度的恒定温度下；(b)沿着密封压机的密封平面定位基底；(c)在基底压缩屈服强度的大约50％至100％的压力下，将正向模具和反向模具的密封表面压入样品处理装置的基底中，持续足够的时间，以将热量通过基底的外表面传递至基底的内表面，从而形成部分熔融的密封件；以及(d)撤回正向模具和反向模具，并使正向模具和反向模具保持无张力，以允许部分熔融的密封件冷却且收缩。在一些实施方案中，当前公开文本中的密封装置用于在样品处理装置中产生易碎的密封件。在一些实施方案中，一对等温加热模具被保持在比管基底的熔融温度低约15℃至25℃的恒定温度下。在一些实施方案中，基底包括聚丙烯，并且将一对等温加热模具保持在大约105℃至115℃的恒定温度下。在一些实施方案中，以大约300psi至600psi的压力将正向模具和反向模具的密封表面压入基底。在一些实施方案中，将正向模具和反向模具压靠在基底上约3秒至6秒。
附图说明
图1A是位于分析仪内部的样品管的截面图。
图1B是样品管的示例性实施方案的透视图。
图2是处于关闭位置的密封压机的截面图。
图3A是在将样品处理装置插入其中之前的处于打开位置的密封压机的截面图。
图3B是在样品处理装置插入其中时处于关闭位置的密封压机的截面图。
具体实施方式
除非本文另有定义，否则与本公开文本结合使用的科学术语和技术术语应具有本领域普通技术人员通常理解的含义。此外，除非上下文另外要求，否则单数术语应包括复数并且复数术语应包括单数。冠词“一个”和“一种”在本文中用于指代冠词的语法对象中的一个或多个(即，至少一个)。举例来说，“一个元件”是指一个元件或不止一个元件。
本文描述的方法用于制造被配置用于执行核酸扩增技术的样品处理装置。可以通过使用以下示例性方法中的至少一种来扩增核酸，来进一步处理从生物样品中提取的核酸：聚合酶链反应(PCR)、滚环扩增(RCA)、连接酶链反应(LCR)、转录介导的扩增(TMA)、基于核酸序列的扩增(NASBA)、以及链置换扩增反应(SDAR)。在一些实施方案中，从生物体提取的核酸可以是核糖核酸(RNA)，并且其处理可以包括使用诸如Tth聚合酶和Taq聚合酶或逆转录酶和Taq聚合酶之类的酶的组合的偶联的逆转录和聚合酶链反应(RT-PCR)。在一些实施方案中，可以使用T4 DNA连接酶或AmpligaseTM和诸如DNA靶或RNA靶之类的引导核酸，将有缺口的环形核酸探针环化，随后在体外选择过程之后检测封闭的环化探针的形成。这种检测可以使用本领域的技术人员已知的酶通过PCR、TMA、RCA、LCR、NASBA或SDAR进行。
在示例性实施方案中，可以通过使用荧光标记的核酸探针或DNA嵌入染料以及分子分析仪中的光度计或电荷耦合元件来实时检测核酸的扩增，以在核酸扩增期间检测荧光的增加。这些荧光标记的探针使用本领域技术人员众所周知的检测方案(即，TaqManTM、分子信标TM、荧光共振能量转移(FRET)探针、scorpionTM探针)，并且通常使用荧光猝灭以及释放从一个报告基因转移到另一个报告基因的猝灭或荧光能量，以检测特定核酸的合成或存在。
在一个实施方案中，本文公开的方法用于制造包括自含式微尺度到宏尺度的通道、腔室、储器、检测区域和处理区域的装置。所述装置可以是药筒、装置、容器或小袋，例如如美国专利号6,440,725、6,783,934、6,818,185、6,979,424、8,580,559以及8,940,526中所描述的，以及诸如可从Cepheid Corp.,Idaho Technology,Inc.和/或BiofireDiagnostics,Inc获得的设备。
在一个具体的实施方案中，本文所述的方法用于在诸如美国专利号7,718,421中所述的样品处理装置中形成密封件。诸如在美国专利号7,718,421中所描述的分段装置提供了用于接收、存储、处理和/或分析生物样品的方便的器皿。在某些实施方案中，分段管促进了涉及多个处理步骤的样品处理协议。在某些实施方案中，将试剂引入管的一个分段中，将样品收集在所述管中，并且然后将所述管放置在可以处理所述管及其所容纳之物的分析仪中，以处理样品。
样品处理装置的一个实施方案在图1A至图1B中示出。样品处理装置100包括框架101和基底102，基底102包括2个或更多个分段110、120、130、140、150、160、170和/或180的布置。分段中的每一个通过可断裂的或易碎的密封件(例如，210、220、230、240、250、260和/或270)分开。在图1A至图1B中描述的实施方案中，分段的布置是线性的，并且基底以管的形式被提供，所述管具有中空中心、外表面和面向所述管的所述中空中心的内表面。然而，还可以设想替代的布置，并且本文描述的方法和装置可以用于创建具有以非线性布置分隔的基底的样品处理装置。
在一个特定的实施方案中，图1A至图1B中描述的线性布置促进了样品以及所产生的废物和靶以受控的方式通过管的运动。例如，可以通过管的第一分段110中的第一开口190输入生物样品。此后，可以在朝向相对末端推动靶的同时，将来自处理过的样品的废物朝向第一开口移回，从而最小化了由可能已附着在管壁上的反应抑制剂对所述靶的污染，并且将所述靶限制在管的干净分段，所述干净分段可以包含用于靶的进一步操作的合适试剂。一些实施方案可以使用多个至少三个分段，每个分段包含至少一种试剂。在一些实施方案中，这些分段可以按以下顺序包含试剂：第二分段中的试剂可以是裂解试剂、稀释缓冲剂或洗涤缓冲剂、或者基底；第三分段中的试剂可以是基底、裂解试剂、洗涤缓冲剂或者中和试剂；第四分段中的试剂可以是洗涤缓冲剂、悬浮缓冲剂、洗脱试剂、或者核酸扩增及检测试剂。在一些实施方案中，三个分段可以连续地布置，而在其他实施方案中，这三个分段可以由它们之间的另一个分段或多个分段分开。
在特定实施方案中，可以将一个或多个易碎的或易破损的密封件，例如210，合并到样品处理设备的基底中以选择性地关闭和打开样品处理装置的基底中的腔室，以允许将存储在第一腔室(例如，110)中的试剂引入到管中的邻近的腔室(例如，120)中。在一些实施方案中，可提供易破损的密封件和压力门的组合以用于将试剂引入端口的内容物转移至邻近分段。
易碎密封件是容易被破损、被撕裂或被切割的密封件，包括但不限于泡罩包装或箔密封件。在一个实施方案中，易碎密封件是可重新密封的或自密封的。易碎密封件被合并到样品处理设备的基底中，以防止在存储过程中定位在给定分段中的试剂混合，但是在测定过程期间，当设备的一个或多个分段的外表面上选择性地施加适当的压力时，所述易碎密封件可以被戳破或者被剥落以允许分段之间混合，而不会破坏基底的外表面。
在图2、图3A和图3B中展示了用于在样品处理装置中产生易碎密封件的装置。装置包括密封压机400，密封压机400包括一对等温加热模具401，该对模具包括正向模具402和反向模具403，其中，各自包括彼此相对并且适于将沿着密封平面406放置的样品处理装置固持在其间的密封表面(分别为404、405)。在特定实施方案中，正向模具和反向模具各自独立地放置在正向对准滑块407和反向对准滑块408上并且分别由其控制，并且正向对准滑块和反向对准滑块连接到致动器409，致动器409提供用于使正向模具和反向模具同步运动的推力。
在一个实施方案中，正向对准滑块和反向对准滑块中的每一个分别安装到正向卡钳臂410和反向卡钳臂411，并且正向卡钳臂和反向卡钳臂各自安装到连接到致动器409的机械联动装置412。在本实施方案中，机械联动装置包括杠杆413，其第一连杆414和第二连杆415分别附接至杠杆413。杠杆413包括中心枢轴点416以及顶端和底端(分别为413a、413b)。第一连杆414包括近端和远端(分别为414a、414b)，并且第一连杆414在远端414b处枢转地连接至杠杆413的顶端413a，并且枢转地连接至反向卡钳臂411的底端411a。同样地，第二连杆415包括近端和远端(分别为415a、415b)，并且第二连杆415在近端415a处枢转地连接至杠杆413b的底端，并且在近端415b处枢转地连接到正向卡钳臂410的底端410a。此外，第一卡钳臂410的上部部分410b和第二卡钳臂411的上部部分411b分别连接到正向对准滑块407和反向对准滑块408。
此外，密封压机还包括独立的正向模具控制机构419，正向模具控制机构419连接到正向模具并且包括与补充的正向模具线性滑块421和独立的正向模具控制器422连接的正向模具驱动器420。正向模具控制机构从正向卡钳臂独立地致动正向模具，从而调节模具密封表面之间的接触压力。
实际上，通过将密封模具加热到低于样品处理装置的基底的熔融温度的恒定温度，来在样品处理装置中产生易碎的密封件。如果基底包括熔融温度为约125℃至135℃的塑料(例如,聚丙烯)，则将密封模具加热到低于熔融温度约15℃至25℃的恒定温度(即，105℃至115℃)。一旦模具达到期望的温度，在操作期间就将所述模具保持在该温度。然后沿着密封压机的在正向模具404与反向模具405之间的密封平面406定位样品处理装置的基底(例如，图3A至图3B中的423)。正向模具和反向模具中的每一个都朝向密封平面移动，反向模具朝向密封平面朝向致动器409移动，而正向模具朝向密封平面远离致动器409移动。一对模具的密封表面在等于或低于基底的压缩屈服强度的压力下被压靠在基底的外表面。例如，压力是基底的压缩屈服强度的大约50％至100％。如果基底是塑料，例如聚丙烯，则压缩屈服强度为约300psi至600psi，并且因此，压力大约为150psi至600psi。一对模具的密封表面被压靠在外表面上(例如，如图3B中所示出的)持续足够的时间(例如，3秒至6秒)，以将热量通过外表面传递到基底的内表面，从而在基底的内表面之间形成部分熔融的密封件。从基底的外表面撤回加热模具，并且将一对模具无张力地保持与基底分开，以允许基底中的部分熔融的密封件冷却且收缩，从而形成易碎的密封件。
可以重复过程以在样品处理装置的基底中产生一个或多个附加的易碎密封件。例如，可以通过在样品处理装置中的中空基底的基座放置一定体积的试剂来启动过程，并且然后将样品处理装置放置在本文所述的且在图2和图3A至图3B中展示的装置中，并且在基底上的基座中的试剂的体积上方产生易碎的密封件，从而在基底中的基座和第一易碎密封件之间形成腔室。随后，将额外体积的试剂(相同或不同的试剂)分配到易碎密封件上方的中空管中，并再一次重复过程以创建额外的易碎密封件和第一易碎密封件与所述额外的易碎密封件之间的基底中的额外的腔室。

Claims (5)

1.一种密封压机，包括一对等温加热模具，所述一对等温加热模具包括正向模具和反向模具，所述正向模具和所述反向模具各自包括彼此相对且适于将沿着密封平面定位的样品处理装置固持在其间的密封表面，其中，所述正向模具和所述反向模具分别独立地定位在正向对准滑块和反向对准滑块上并且分别由其控制，所述正向对准滑块和所述反向对准滑块连接到致动器，所述致动器提供用于使所述正向模具和所述反向模具同步运动的推力， 其中，所述正向对准滑块和所述反向对准滑块分别安装到正向卡钳臂和反向卡钳臂，并且所述正向卡钳臂和所述反向卡钳臂分别安装到与所述致动器连接的机械联动装置，其中，所述机械联动装置包括杠杆、第一连杆和第二连杆，其中，所述杠杆包括中心枢转点以及第一末端和第二末端，其中，所述第一连杆枢转地连接到所述杠杆的所述第一末端并且枢转地连接到所述反向卡钳臂的一个末端，其中，所述第二连杆枢转地连接到所述杠杆的所述第二末端并且枢转地连接到正向卡钳臂的一个末端，其中，所述正向卡钳臂和所述反向卡钳臂分别连接至所述正向对准滑块和所述反向对准滑块， 进一步包括独立的正向模具控制机构，所述机构连接到所述正向模具，并且包括连接到补充性正向模具线性滑块和独立的正向模具控制器的正向模具驱动器，其中，所述机构从所述正向卡钳臂独立地致动所述正向模具，从而调节所述密封表面之间的接触压力。
2.根据权利要求1所述的密封压机，其中，所述一对等温加热模具被配置用于被加热到恒定温度，所述恒定温度低于所述样品处理装置的基底的熔融温度。
3.根据权利要求2所述的密封压机，其中，所述基底包括聚丙烯，并且所述一对等温加热模具被加热到105ºC至115ºC。
4.一种在包括具有外表面和内表面的中空管基底的样品处理装置中产生易碎密封件的方法，其中，所述方法是由包括一对等温加热模具的密封压机来执行，所述一对等温加热模具包括正向模具和反向模具，所述正向模具和所述反向模具各自包括彼此相对且适于将沿密封平面定位的样品处理装置保持在其间的密封表面，其中，所述正向模具和反向模具各自独立地定位在正向对准滑块和反向对准滑块上并且分别由其控制，所述正向对准滑块和所述反向对准滑块连接到致动器，所述致动器提供用于使所述正向模具和所述反向模具同步运动的推力，所述方法包括：
(a) 将所述一对等温加热模具维持在低于所述样品处理装置的基底的熔融温度的恒定温度下；
(b) 沿着所述密封压机的所述密封平面定位所述基底；
(c) 在所述基底的压缩屈服强度的50％至100％的压力下，将所述正向模具和所述反向模具的所述密封表面压入所述样品处理装置的所述基底中，持续足够的时间，以将热量通过所述基底的外表面传递到所述基底的内表面，从而形成部分熔融的密封件；以及
(d) 撤回所述正向模具和所述反向模具，并且使所述正向模具和所述反向模具保持无张力，以使所述部分熔融的密封件冷却且收缩。
5.根据权利要求4所述的方法，其中， (a) 将所述一对等温加热模具保持在比所述管基底的所述熔融温度低15ºC至25ºC的恒定温度下；和/或 (b) 所述基底包括聚丙烯，并且将所述一对等温加热模具保持在105ºC至115ºC的恒定温度；和/或 (c) 以300 psi至600psi的压力将所述正向模具和所述反向模具的所述密封表面压入所述基底中；和/或 (d)将所述正向模具和所述反向模具压靠在所述基底上持续3秒至6秒。
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