WO2006047912A1 - A centrifugal tube for cleaning microarray chips - Google Patents

A centrifugal tube for cleaning microarray chips Download PDF

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Publication number
WO2006047912A1
WO2006047912A1 PCT/CN2004/001342 CN2004001342W WO2006047912A1 WO 2006047912 A1 WO2006047912 A1 WO 2006047912A1 CN 2004001342 W CN2004001342 W CN 2004001342W WO 2006047912 A1 WO2006047912 A1 WO 2006047912A1
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WIPO (PCT)
Prior art keywords
cleaning
tube
chip
microarray
tube body
Prior art date
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Ceased
Application number
PCT/CN2004/001342
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English (en)
French (fr)
Inventor
Feijun Xian
Yimin Sun
Xiaoxiang Zhu
Wanli Xing
Yan Wang
Jing Cheng
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Tsinghua University
CapitalBio Corp
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Tsinghua University
CapitalBio Corp
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Publication of WO2006047912A1 publication Critical patent/WO2006047912A1/zh
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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L13/00—Cleaning or rinsing apparatus
    • B01L13/02—Cleaning or rinsing apparatus for receptacle or instruments
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/06—Auxiliary integrated devices, integrated components
    • B01L2300/0609—Holders integrated in container to position an object
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0809—Geometry, shape and general structure rectangular shaped
    • B01L2300/0822—Slides
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • 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
    • B01L3/508—Rigid containers without fluid transport within
    • B01L3/5082—Test tubes per se
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/00029—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor provided with flat sample substrates, e.g. slides
    • G01N2035/00099—Characterised by type of test elements
    • G01N2035/00158—Elements containing microarrays, i.e. "biochip"
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N2035/00465—Separating and mixing arrangements
    • G01N2035/00564—Handling or washing solid phase elements, e.g. beads

Definitions

  • the invention relates to a microarray chip cleaning and a centrifuge tube.
  • Microarray Chips are widely used in biological, medical, food safety and related fields as a fast and efficient inspection tool. Microarray chips are used to place and fix a variety of probes (such as nucleic acids, proteins, or tissues) in a regular row and column arrangement. The current substrate is mostly standard slides (size is about 25). Mmx75mmx'l mm). The basic steps for detection and analysis using microarray chips include: sample preparation, biochemical reactions, and results detection. In the process of biochemical reaction of the microarray chip, chip cleaning and centrifugation are often required.
  • the cleaning method of the microarray chip is generally carried out in a cleaning container (such as a common plastic box, a beaker) on a shaker.
  • a cleaning container such as a common plastic box, a beaker
  • the specific operation process is as follows: Place the chip in a cleaning container containing the cleaning liquid, place the cleaning container on the shaker, turn on the power of the shaker for oscillation cleaning, and remove the chip from the cleaning container after the cleaning is completed, and perform centrifugation.
  • the commonly used microarray chip centrifugation methods mainly have the following two methods: First, the cleaned chip (standard slide) is inserted into a 50 ml ordinary plastic centrifuge tube and centrifuged on a conventional centrifuge; Second, the application of TeleChem (Http: //www.array it com) Developed a microarray chip centrifuge for centrifugation of a standard slide. The cleaned slides are inserted into the slide holder on the centrifuge and centrifuged.
  • the object of the present invention is a microarray chip cleaning and centrifuge tube capable of both cleaning and centrifuging functions.
  • the present invention uses the following technical solution: a microarray chip cleaning, a centrifuge tube, comprising a tube body, the inner side wall of the tube body is provided with a pair of chip positioning grooves, the tube of the tube body The bottom inner side is provided with at least one chip support rib protruding from the inner surface.
  • the portion of the chip positioning groove near the nozzle is configured as a bell mouth.
  • the microarray chip cleaning and centrifuge tube further includes a tube cover, the tube cover and the tube body Connected by thread. A sealing ring is also disposed in the tube cover.
  • the longitudinal section of the tubular body near the bottom portion of the tube is trapezoidal, forming a tapered liquid collecting space, which ensures the centrifugal effect of the chip.
  • the tubular body surface of the tubular body is a flat surface or a curved surface; and the tubular body is preferably cylindrical.
  • the inner wall of the tube body is a smooth surface, a calender surface or a sand surface; wherein, in the light surface, an auroral surface is preferred.
  • a fixed volume of cleaning liquid is added, and the outer wall of the tube body is further provided with a volume scale mark.
  • the invention has the following advantages due to adopting the above technical solutions: 1) chip support ribs and chip sets
  • the setting of the position groove and the chip guiding groove facilitates the loading and unloading operation of the chip, and the position of the chip is unchanged in the tube before and after centrifugation, thereby avoiding the phenomenon of "sinking" of the chip when the ordinary 50 ml centrifuge tube is used for centrifugation.
  • the setting of the tube cover and the sealing ring makes the invention have good sealing performance, which can be used not only for chip cleaning, centrifugation, but also for chip storage; 3)
  • the invention can be operated by a conventional centrifuge without using a dedicated centrifugal device and is easy to operate.
  • FIG. 1A is a schematic structural view of the present invention
  • Figure 1B is a schematic view showing the internal structure of the present invention.
  • Figure 1C is a cross-sectional view taken along line A-A of Figure 1A; 2A is a schematic structural view of the present invention after loading a chip;
  • Figure 2B is a cross-sectional view taken along line B-B of Figure 2A;
  • Figure 3A is a schematic view of the structure of the present invention with a tube cover and a sealing ring;
  • Figure 3B is a cross-sectional view taken along line C-C of Figure 3A;
  • FIG. 4A is a schematic structural view of a cleaning tube having a cylindrical body
  • 4B is a schematic view showing the internal structure of a cleaning tube having a cylindrical body
  • Figure 5A is a result scan of a chip using conventional cleaning and centrifugation
  • FIG. 5B is a result scan of a chip for performing chip cleaning and centrifugation using the present invention
  • FIG. 5C is a comparison diagram of signal values of scan results
  • Figure 5D is a comparison of the coefficient of variation values.
  • Example 1 Microarray chip cleaning, centrifuge tube
  • the microarray chip cleaning and the centrifuge tube comprise a tube body 1.
  • the inner wall of the tube bottom is provided with a chip supporting rib 2, and the side wall of the tube body has a chip positioning groove 3, the chip
  • the support rib 2 and the chip positioning groove 3 are arranged to ensure that the position of the chip is unchanged in the tube before and after centrifugation, thereby avoiding the phenomenon of "sinking" of the chip when the ordinary 50 ml centrifuge tube is used for centrifugation;
  • the side wall of the nozzle also has a chip guiding groove 4 with a certain curvature.
  • the chip guiding groove 4 has a "horn" shape and is integrated with the chip positioning groove 3; in order to ensure the centrifugal effect of the chip, the tube body is close to The longitudinal section of the bottom portion of the tube is trapezoidal, and a liquid collecting space 5 having a certain taper is formed at the bottom of the tube.
  • a volume scale mark 6 is arranged on the outer wall of the pipe body 1 to facilitate adding a certain volume of cleaning liquid when cleaning the chip;
  • the shape of the surface 8 of the pipe body and the lower surface 9 of the pipe body may be a flat shape, which is convenient for cleaning the pipe. It can also be used to mark and paste labels.
  • the surface of the inner wall of the pipe body 1 may be an auroral surface, a glossy surface, a calender surface, or a sand surface, etc. In order to prevent adhesion of biological small molecules, the surface of the inner wall is preferably an auroral surface.
  • the material of the tube body 1 may be polypropylene, polycarbonate, plexiglass, polystyrene, ABS, polyethylene, polyvinyl chloride, polyoxymethylene, etc., and the light transmittance may be opaque, translucent, or transparent, and ⁇ Processing by conventional injection molding or blow molding processes.
  • FIGS. 2A and 2B are schematic views showing the structure of the cleaning and centrifugation tube loading chip 10.
  • the chip 10 is placed in the chip positioning groove 3, and one end thereof is in contact with the chip supporting rib 2.
  • the cleaning tube is not only It can be used to load ordinary chips, and can load multi-sample fence type chips, which can be centrifuged on a conventional centrifuge.
  • 3A and 3B are schematic views showing the structure of the cleaning tube of the present invention equipped with a tube cover and a sealing ring.
  • the present invention may also be provided with a tube cover 11 and a sealing ring 12, and the tube cover 11 and the tube body 1 are connected by a thread 13, and the sealing ring 12 is mounted in the tube cover 11.
  • the material of the tube cover 11 can be the same as that of the tube body 1, such as polypropylene, polycarbonate, plexiglass, polystyrene, ABS, polyethylene, polyvinyl chloride, polyoxymethylene, etc., and the light transmittance can be opaque. , translucent, or transparent, can be processed by conventional injection molding or blow molding processes.
  • the sealing ring 12 can be processed by using silicone rubber, natural rubber, nitrile rubber, urethane rubber, neoprene rubber, fluororubber rubber, ethylene propylene rubber or the like.
  • the common processing techniques include vulcanization or stamping, and the light transmittance is opaque. Translucent, 'or transparent, etc.
  • FIGS. 4A and 4B are schematic views showing the structure of a cleaning tube having a cylindrical body.
  • the tube body surface 8 of the tube has a cylindrical surface shape similar to the outer surface of the 50 ml centrifuge tube.
  • Example 2 the application of the invention in the field of protein chip detection
  • FIG. 5A is a result scan of a chip using conventional cleaning and centrifugation
  • FIG. 5B is a result scan of a chip for performing chip cleaning and centrifugation using the present invention
  • FIG. 5C is a comparison chart of signal values of the scan result
  • FIG. 5D is a coefficient of variation value. Comparison chart.
  • the difference between the probe signal intensity value and the coefficient of variation of the chip after cleaning and centrifugation according to the present invention is within an acceptable error range, indicating that the present invention can be used for cleaning, centrifuging, and microchip chip. It is very convenient to pick and place from the cleaning tube.
  • Microarray chip cleaning and centrifugal tube structure are simple, convenient and practical, and will be widely used in cleaning and centrifugation of microarray chips.

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  • Health & Medical Sciences (AREA)
  • Clinical Laboratory Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Centrifugal Separators (AREA)
  • Sampling And Sample Adjustment (AREA)

Description

微阵列芯片清洗、 离心管 技术领域
本发明涉及一种微阵列芯片清洗、 离心管。
背景技术
微阵列芯片 （Microarray Chip)作为一种快速、 高效的检测工具而广泛 应用于生物、 医学、 食品安全及相关领域。 微阵列芯片是将多种探针 （如核 酸、 蛋白、 或组织） 以规则的行与列的排列方式点制并固定于基片上， 目前 的基片多采用标准载玻片 （尺寸约为 25 mmx75mmx'l mm) 。 利用微阵列芯 片进行检测、 分析的基本步骤包括： 样品制备、 生化反应和结果检测。 在微 阵列芯片完成生化反应的过程中， 往往需要进行芯片清洗及离心。
目前， 微阵列芯片的清洗方法， 一般是把芯片放入清洗容器（如： 普通 塑料盒、 烧杯） 中， 在摇床上进行。 具体操作过程是： 将芯片放于盛有清洗 液的清洗容器中， 将清洗容器放于摇床上， 打开摇床电源进行振荡清洗， 清 洗完成后从清洗容器中取出芯片， 进行离心。
常用的微阵列芯片离心方法主要有以下两种方式： 第一种， 将清洗后的 芯片（标准载玻片）插入 50ml普通塑料离心管， 在常规离心机上进行离心； 第二种， 应用 TeleChem公司 （Http: //www. array it com) 开发的微阵列芯片 离心机， 进行 1片标准载玻片的离心。 将清洗后的载玻片插入该离心机上的 载玻片固定架， 便可离心。
对于第一种方式，这些 50ml离心管用于芯片的离心时，存在着如下问题: 1 ) 50ml离心管一般较长，而且带一定的锥度（越往底部管体截面直径越小）， 造成芯片离心后卡在离心管底部， 不容易取出; 2)当芯片宽度公差较大（即 宽度较宽）时， 经常不能顺利插入离心管； 3 )芯片的清洗与离心分别在不同 容器内完成， 整个过程需要手动转移芯片， 不仅操作繁琐， 而且在转移过程 中具有安全隐患。
对于第二种方式， 需要购买专用离心机， 成本较高； 而且离心效率低， 一次只能离心 1片芯片； 同时， 芯片的清洗与离心也在不同容器内完成， 整 个过程需要手动转移芯片， 也存在操作繁琐等问题。
发明公开 本发明的目的是能同时具备清洗和离心功能的微阵列芯片清洗、离心管。 为实现上述目的， 本发明釆用如下技术方案： 一种微阵列芯片清洗、 离 心管， 包括一管体， 所述管体的内侧壁上设有一对芯片定位凹槽， 所述管体 的管底内侧设有至少一道突出于内表面的芯片支撑筋。
为了便于取放芯片， 所述芯片定位凹槽靠近管口的部分设为喇叭口状。 为了能保证芯片在用于清洗时的水密封性， 以及在用于存储芯片时的气 密封性， 所述微阵列芯片清洗、 离心管还包括一管盖， 所述管盖与所述管体 通过螺紋连接。 所述管盖内还设有密封圈。
作为本发明的一种改进， 所述管体靠近管底部分的纵切面为梯形， 形成 带锥度的液体收集空间， 可以确保芯片的离心效果。
在本发明中， 所述管体的管身表面为平面或曲面； 所述管体优选为圆柱 形。
为防止粘附生物小分子， 所述管体的内壁为光面、 压光面或砂面； 其中， 光面中优选的是极光表面。
以方便清洗芯片时， 加入固定体积的清洗液， 所述管体的外壁上还设有 容积刻度标识。
在使用时， 将芯片放于管内， 加入清洗液， 旋入带有密封圈的管盖， 放 于水平摇床进行清洗， 清洗完成后倒掉清洗液， 然后用常规离心机离心， 即 可完成芯片的清洗、 离心。
本发明由于采用以上技术方案， 具有如下优点： 1 )芯片支撑筋、 芯片定
' 位凹槽以及芯片导向槽的设置， 方便了芯片的装载与取出操作， 并使芯片离 心前后在管内位置不变， 避免了应用普通 50ml离心管进行离心时芯片的 "下 沉"现象， 可以在一个管内进行清洗和离心操作； 2) 管盖、 密封圈的设置， 使得本发明具有良好的密封性， 不仅能用于芯片的清洗、 离心， 而且也可以 用于芯片的存储； 3)本发明可以采用常规的离心机进行操作， 不需要使用专 用的离心设备， 操作方便。
附图说明
图 1A为本发明的结构示意图；
图 1B为本发明的内部结构示意图；
图 1C为图 1A的 A- A剖面图； 图 2A为装载芯片后的本发明结构示意图；
图 2B为图 2A的 B- B剖面图；
图 3A为带有管盖和密封圈的本发明结构示意图；
图 3B为图 3A的 C- C剖面图；
图 4A为管身为圆柱形的清洗管的结构示意图；
图 4B为管身为圆柱形的清洗管的内部结构示意图；
图 5A为利用常规清洗与离心的芯片的结果扫描图；
图 5B为利用本发明进行芯片清洗与离心的芯片的结果扫描图； 图 5C为扫描结果信号值的比较图；
图 5D为变异系数值的比较图。
实施发明的最佳方式
实施例 1、 微阵列芯片清洗、 离心管
见图 1A、 图 IB和图 1C， 本实施例中微阵列芯片清洗、 离心管， 包括管 体 1， 管底内壁上设有芯片支撑筋 2， 管体侧壁有芯片定位凹槽 3， 芯片支撑 筋 2与芯片定位凹槽 3的设置， 确保了芯片离心前后在管内位置不变， 避免 了应用普通 50ml离心管进行离心时芯片的 "下沉"现象； 为了方便芯片的取 放， 在靠近管口的侧壁上还有带一定弧度的芯片导向槽 4， 该芯片导向槽 4 呈 "喇叭口 "状， 并与芯片定位凹槽 3连成一体； 为了确保芯片的离心效果， 管体靠近管底部分的纵切面为梯形， 在管底处形成具有一定锥度的液体收集 空间 5。
在管体 1外壁上设有容积刻度标识 6， 以方便清洗芯片时， 加入一定体 积的清洗液； 管体管身上表面 8、管身下表面 9的形状可以是平面形状， 方便 清洗管的平放， 而且还可以用来进行标记及粘贴标签。 管体 1内壁的表面可 以是极光表面、 光面、 压光面、 或砂面等； 为防止粘附生物小分子， 内壁表 面优选极光表面。
管体 1的制造材料可以是聚丙烯、聚碳酸酯、有机玻璃、聚苯乙烯、 ABS、 聚乙烯、 聚氯乙烯、 聚甲醛等， 其透光性可为不透明、 半透明、 或透明， 可 以釆用常规的注塑或吹塑等工艺进行加工。
图 2A和图 2B为本发明清洗、离心管装载芯片 10后的结构示意图， 芯片 10放置于芯片定位凹槽 3中， 其一端与芯片支撑筋 2接触。 该清洗管不但可 以用于装载普通芯片， 而且可以装载多样品围栏型芯片， 可在常规离心机上 离心。
图 3A和图 3B为装配有管盖、 密封圈的本发明清洗管的结构示意图。 为 了保证在清洗芯片 10时的水密封性， 以及存储芯片 10时的气密封性， 本发 明还可以设置有管盖 11、 密封圈 12， 管盖 11与管体 1通过螺紋 13连接， 密 封圈 12安装于管盖 11内。
管盖 11的制造材料可以与管体 1相同，如聚丙烯、聚碳酸酯、有机玻璃、 聚苯乙烯、 ABS、 聚乙烯、 聚氯乙烯、 聚甲醛等均可， 其透光性可为不透明、 半透明、 或透明， 可以采用常规的注塑或吹塑等工艺进行加工。
密封圈 12可以采用硅橡胶、天然橡胶、 丁腈橡胶、 聚氨酯橡胶、 氯丁橡 胶、 氟橡胶、 乙丙橡胶等材料进行加工制造， 常用加工工艺有硫化或冲压等， 其透光性为不透明、 半透明、' 或透明等。
图 4A和图 4B为管身为圆柱形的清洗管的结构示意图。该管的管身上表 面 8、 管身下表面 9的形状为圆柱面， 形状类似于 50ml离心管的外表面。
实施例 2、 本发明在蛋白芯片检测领域中的应用
利用以标准载玻片为基片的微阵列蛋白芯片进行动物源性食品中兽药残 留物 （恩诺沙星、 新霉素、 磺胺、 链霉素） 的检测， 芯片的制作、 检测等均 采用本领域的常规方法进行。 检测过程中， 釆用相同的芯片格式 （10孔围栏 芯片与围栏芯片盖片） 、 相同的样品、 相同的探针、 相同的免疫反应条件、 相同的扫描参数， 仅仅是芯片免疫反应后分别采用两种不同的清洗、 离心方 法：
1 )常规的芯片清洗与离心方法:将芯片放于盛有清洗液的普通塑料盒中， 将塑料盒放于水平摇床进行清洗， 清洗完成后用从塑料盒中取出芯片， 放于 50ral清洗管， 用常规离心机 （飞鸽 TDL-5 ) 离心。
2)利用本发明进行芯片清洗与离心的方法： 将芯片放于微阵列芯片清洗 管管体， 加入清洗液， 旋入装有密封圈的管盖， 放于水平摇床进行清洗， 清 洗完成后倒掉清洗液， 然后用常规离心机 （飞鸽 TDL-5 ) 离心。
图 5A为利用常规清洗与离心的芯片的结果扫描图， 图 5B为利用本发明 进行芯片清洗与离心的芯片的结果扫描图， 图 5C 为扫描结果信号值的比较 图， 图 5D为变异系数值的比较图。 从图中可见， 与常规的芯片清洗、 离心的 方法相比， 采用本发明进行清洗、 离心后的芯片， 其探针信号强度值、 变异 系数差值均在可接受的误差范围内，表明本发明可以用于微阵列芯片的清洗、 离心， 芯片从清洗管中的取放均非常方便。
工业应用
微阵列芯片清洗、 离心管结构简单， 方便实用， 在微阵列芯片的清洗和 离心中将得到广泛应用。

Claims

极 利 要 求 书
1、一种微阵列芯片清洗、 离心管， 包括一管体， 其特征在于： 所述管体 的内侧壁上设有一对芯片定位凹槽， 所述管体的管底内侧设有至少一道突出 于内表面的芯片支撑筋。
2、 根据权利要求 1所述的微阵列芯片清洗、 离心管， 其特征在于： 所述 芯片定位凹槽靠近管口的部分为喇叭口状。
3、 根据权利要求 1或 2所述的微阵列芯片清洗、 离心管， 其特征在于： 所述微阵列芯片清洗、 离心管还包括一管盖， 所述管盖与所述管体通过螺紋 连接。
4、 根据权利要求 3所述的微阵列芯片清洗、 离心管， 其特征在于： 所述 管盖内还设有密封圈。
5、 根据权利要求 1或 2所述的微阵列芯片清洗、 离心管， 其特征在于： 所述管体靠近管底部分的纵切面为梯形。
6、 根据权利要求 5所述的微阵列芯片清洗、 离心管， 其特征在于： 所述 管身表面为平面或曲面。
7、 根据权利要求 6所述的微阵列芯片清洗、 离心管， 其特征在于： 所述 管体为圆柱形。
8、 根据权利要求 1或 2所述的微阵列芯片清洗、 离心管， 其特征在于： 所述管体的内壁为光面、 压光面或砂面。
9、 根据权利要求 1或 2所述的微阵列芯片清洗、 离心管， 其特征在于： 所述管体的外壁上还设有容积刻度标识。
PCT/CN2004/001342 2004-11-05 2004-11-23 A centrifugal tube for cleaning microarray chips Ceased WO2006047912A1 (en)

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