WO2021114086A1 - Cell counting device - Google Patents

Cell counting device Download PDF

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Publication number
WO2021114086A1
WO2021114086A1 PCT/CN2019/124300 CN2019124300W WO2021114086A1 WO 2021114086 A1 WO2021114086 A1 WO 2021114086A1 CN 2019124300 W CN2019124300 W CN 2019124300W WO 2021114086 A1 WO2021114086 A1 WO 2021114086A1
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cell
observation
tube
counting device
pool
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PCT/CN2019/124300
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French (fr)
Chinese (zh)
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张豪
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苏州生动细胞生物科技有限公司
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Priority to PCT/CN2019/124300 priority Critical patent/WO2021114086A1/en
Publication of WO2021114086A1 publication Critical patent/WO2021114086A1/en

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/34—Measuring or testing with condition measuring or sensing means, e.g. colony counters
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/1031—Investigating individual particles by measuring electrical or magnetic effects
    • G01N15/12—Investigating individual particles by measuring electrical or magnetic effects by observing changes in resistance or impedance across apertures when traversed by individual particles, e.g. by using the Coulter principle

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  • This application relates to the field of cell culture technology, and in particular to a cell counting device.
  • the existing cell counter When the existing cell counter performs the cell counting operation, firstly, the cell liquid is manually taken and the cell liquid is dropped on the counting plate, and then the counting plate is inserted into the counter (counter) to read the result information of the cell number.
  • Using this open method of manual liquid extraction requires many requirements for the cleanliness of the operating environment and operating steps, which increases the complex steps of sampling, and the existing counting method exposes all cultured cells to the air The cells are easily contaminated; the number of cells in the process of manual liquid extraction will also cause more losses and cause cell waste.
  • the purpose of this application is to overcome the problems that the existing cell counter requires manual sampling, which is easy to cause overall cell culture pollution and complicated operations, and to provide a real-time online cell counting device, including a cell circulation tube, a peristaltic pump, a restrictor valve, an observation block, Counter and display;
  • the cell circulation tube is provided with at least two openings for communicating with the cell culture bag;
  • the observation block is transparent, and an observation pool connected with the cell circulation tube is arranged inside the observation block; the observation pool includes an inlet end and an outlet end;
  • the peristaltic pump is connected to the cell circulation tube;
  • the flow limiting valve is connected to the cell circulation tube and is arranged at the inlet end of the observation block for limiting the thickness of the cell liquid surface entering the observation pool;
  • the counter includes an illuminating device and a microscope arranged at intervals in a longitudinal direction, and the counter is electrically connected with a display; the observation block is located between the illuminating device and the microscope.
  • the cell circulation tube includes two symmetrically arranged tube bodies, one end of the tube body communicates with the inlet end of the observation cell, and one end of the other tube body communicates with the outlet end of the observation cell.
  • the pipe body includes a plurality of straight pipes, elbows and connectors; each straight pipe and/or elbow is connected by the connector.
  • the tube body includes an inner tube and an outer tube; the inner tube is in communication with the observation tank.
  • the thickness of the cell liquid surface in the observation pool is 0-0.5 mm.
  • the thickness of the cell liquid surface in the observation pool is 0.1 mm.
  • observation pool is a U-shaped groove.
  • the beneficial effects of the present application are: through the cooperation of the sealed cell circulation tube and the observation block, the automatic closed circulation counting of the cell fluid is realized, and the cells do not need to be manually sampled and exposed to contamination in the operating environment, and at the same time, they are no longer manual counting. simple.
  • Figure 1 is a schematic diagram of the structure of the cell counting device of the present application.
  • Figure 2 is a schematic diagram of the structure of Figure 1 during use.
  • Fig. 3 is a schematic diagram of the structure of Fig. 1 with the counter and display removed.
  • Fig. 4 is a schematic diagram of the structure of the observation pool in Fig. 3.
  • Fig. 5 is a schematic diagram of the structure in Fig. 3 with the observation block removed.
  • the cell counting device shown in Figures 1 to 5 includes a cell circulation tube 1, a peristaltic pump 2, a flow limiting valve 3, an observation block 4, a counter 5 and a display 6;
  • the cell circulation tube 1 is provided with at least two openings for communicating with the cell culture bag 8;
  • the observation block 4 is transparent, preferably a non-toxic transparent plastic material, and is provided with an observation pool 7 communicating with the cell circulation tube 1; the observation pool 7 includes an inlet end and an outlet end; the observation pool 7 can be a U-shaped groove, or Can be other shapes.
  • the peristaltic pump 2 is connected to the cell circulation tube 1; the restrictor valve 3 is connected to the cell circulation tube 1 and is arranged at the inlet end of the observation block 4 to limit the thickness of the cell liquid surface entering the observation pool 7; the cell liquid in the observation pool 7
  • the thickness of the surface is 0-0.5mm, preferably 0.1mm, which can ensure that the cells in the cell liquid entering the observation pool 7 are in a single layer without superimposition, which is more conducive to counting.
  • the peristaltic pump 2 and the restrictor valve 3 can be directly selected from the relevant models purchased on the market and used in the cell culture system.
  • the counter 5 includes an illuminating device and a microscope arranged at intervals in the longitudinal direction, and the counter 5 is electrically connected to the display 6; the observation block 4 is located between the illuminating device and the microscope. Through the cooperation of the illumination device and the microscope (electron microscope), the cells in the observation cell 7 can be automatically counted and displayed on the display 6.
  • the counter 5 can also be other existing technologies, such as a method of taking cell images through a camera for counting.
  • the counter 5 of this embodiment is actually an existing product, so the specific implementation or structure will not be described again.
  • the cell circulation tube 1 can be a U-shaped tube.
  • the two ends of the tube are the openings respectively inserted into the cell culture bag 8.
  • a part of the tube is a transparent section and is set between the restrictor valve 3 and the peristaltic pump 2. It is necessary to use the counter 5 to photograph and count the transparent section.
  • the cell circulation tube 1 of this embodiment adopts another form, that is, the cell circulation tube 1 includes two symmetrically arranged tube bodies, as shown in Figures 3 and 5, one end of which is connected to the inlet end of the observation pool 7 , One end of the other tube is connected to the outlet end of the observation tank 7, and the inlet and outlet ends of the observation tank 7 depend on the direction of the peristaltic pump 2.
  • the pipe body may include a plurality of straight pipes 1.1, elbows 1.2, and connectors 1.3; each straight pipe 1.1 and/or elbow 1.2 is connected by the connector 1.3.
  • the length and arrangement of the tube body can be adjusted by the number and matching mode of the straight pipe 1.1 and the elbow 1.2; it can be applied to cell culture bags 8 of different sizes.
  • the tube body in order to reduce cell loss, includes an inner tube and an outer tube; the inner tube is in communication with the observation pool 7.
  • the outer tube protects the inner tube.
  • the straight pipe 1.1 is provided with multiple through holes.
  • the arrangement of the through holes can ensure that cells at different positions in the cell culture bag 8 can enter the observation pool 7 to ensure the accuracy of counting.
  • the cell counting device of this embodiment may further include a sealing plug matched with the cell culture bag 8 to ensure the tightness of the connection between the tube body and the cell culture bag 8.

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Abstract

A cell counting device, comprising a cell circulating tube, a peristaltic pump, a flow limiting valve, an observation block, a counter and a display; the cell circulating tube being provided with at least two openings in communication with a cell culture bag; the observation block being transparent, and an interior thereof being provided with an observation pool in communication with the cell circulating tube; the peristaltic pump and the flow limiting valve being connected to the cell circulating tube; the counter comprising an illumination device and a microscope, disposed vertically at an interval from one another, the counter being electrically connected to the display; the observation block being between the illumination device and the microscope. By means of the combination of the cell circulating tube and the observation block, a cell liquid is circulated and counted in an automatic and sealed fashion, cells are not contaminated or lost, and are not manually counted, and operation is simple.

Description

细胞计数装置Cell counting device 技术领域Technical field
本申请涉及细胞培养技术领域，尤其涉及一种细胞计数装置。This application relates to the field of cell culture technology, and in particular to a cell counting device.
背景技术Background technique
现有的细胞计数器进行细胞计数操作时，首先人工取细胞液并将细胞液滴在计数板上，再把计数板插进计数器(计数仪)读数从而得出细胞数量结果信息。采用这种人工取液的开放式方式，对操作环境的洁净程度和操作步骤要求繁多，增加了抽检的复杂步骤，且现有的这种计数方式，使培养的所有细胞都有暴露在空气中的风险，细胞容易受到污染；在人工取液的过程中细胞的数量也会造成更多的损失，造成细胞浪费。When the existing cell counter performs the cell counting operation, firstly, the cell liquid is manually taken and the cell liquid is dropped on the counting plate, and then the counting plate is inserted into the counter (counter) to read the result information of the cell number. Using this open method of manual liquid extraction requires many requirements for the cleanliness of the operating environment and operating steps, which increases the complex steps of sampling, and the existing counting method exposes all cultured cells to the air The cells are easily contaminated; the number of cells in the process of manual liquid extraction will also cause more losses and cause cell waste.
发明内容Summary of the invention
本申请的目的在于克服现有细胞计数器需要人工取样，易造成整体细胞培养污染，操作复杂的问题，提供一种实时在线细胞计数装置，包括细胞循环管、蠕动泵、限流阀、观察块、计数仪和显示器；The purpose of this application is to overcome the problems that the existing cell counter requires manual sampling, which is easy to cause overall cell culture pollution and complicated operations, and to provide a real-time online cell counting device, including a cell circulation tube, a peristaltic pump, a restrictor valve, an observation block, Counter and display;
所述细胞循环管设有至少两个用于与细胞培养袋连通的开口处；The cell circulation tube is provided with at least two openings for communicating with the cell culture bag;
所述观察块为透明状，其内部设有与细胞循环管连通的观察池；所述观察池包括入口端和出口端；The observation block is transparent, and an observation pool connected with the cell circulation tube is arranged inside the observation block; the observation pool includes an inlet end and an outlet end;
所述蠕动泵与细胞循环管连接；所述限流阀与细胞循环管连接并设于观察块的入口端用于限定进入观察池内的细胞液面厚度；The peristaltic pump is connected to the cell circulation tube; the flow limiting valve is connected to the cell circulation tube and is arranged at the inlet end of the observation block for limiting the thickness of the cell liquid surface entering the observation pool;
所述计数仪包括纵向间隔设置的照明装置和显微镜，所述计数仪与显示器电连接；所述观察块位于所述照明装置和显微镜之间。The counter includes an illuminating device and a microscope arranged at intervals in a longitudinal direction, and the counter is electrically connected with a display; the observation block is located between the illuminating device and the microscope.
进一步，所述细胞循环管包括两根对称设置的管体，其中一个管体的一端与观察池的入口端连通，另一个管体的一端与观察池的出口端连通。Further, the cell circulation tube includes two symmetrically arranged tube bodies, one end of the tube body communicates with the inlet end of the observation cell, and one end of the other tube body communicates with the outlet end of the observation cell.
进一步，所述管体包括多个直管、弯头和连接头；各直管和/或弯头通过连接头连接。Further, the pipe body includes a plurality of straight pipes, elbows and connectors; each straight pipe and/or elbow is connected by the connector.
进一步，所述管体包括内管和外管；所述内管与观察池连通。Further, the tube body includes an inner tube and an outer tube; the inner tube is in communication with the observation tank.
进一步，所述直管上设有多个通孔。Further, a plurality of through holes are provided on the straight pipe.
进一步，所述观察池内的细胞液面厚度为0～0.5mm。Further, the thickness of the cell liquid surface in the observation pool is 0-0.5 mm.
进一步，所述观察池内的细胞液面厚度为0.1mm。Further, the thickness of the cell liquid surface in the observation pool is 0.1 mm.
进一步，所述观察池为U形槽。Further, the observation pool is a U-shaped groove.
本申请的有益效果是：通过密封的细胞循环管和观察块配合，实现了细胞液的自动封闭式循环计数，细胞不用人工取样暴露在操作环境中受到污染，同时也不再是人工计数，操作简单。The beneficial effects of the present application are: through the cooperation of the sealed cell circulation tube and the observation block, the automatic closed circulation counting of the cell fluid is realized, and the cells do not need to be manually sampled and exposed to contamination in the operating environment, and at the same time, they are no longer manual counting. simple.
附图说明Description of the drawings
图1为本申请细胞计数装置的结构示意图。Figure 1 is a schematic diagram of the structure of the cell counting device of the present application.
图2为图1使用过程中的结构示意图。Figure 2 is a schematic diagram of the structure of Figure 1 during use.
图3为图1中去掉计数仪和显示器后的结构示意图。Fig. 3 is a schematic diagram of the structure of Fig. 1 with the counter and display removed.
图4为图3中观察池的结构示意图。Fig. 4 is a schematic diagram of the structure of the observation pool in Fig. 3.
图5为图3中去掉观察块后的结构示意图。Fig. 5 is a schematic diagram of the structure in Fig. 3 with the observation block removed.
图中，1.细胞循环管；1.1.直管；1.2.弯头；1.3.连接头；2.蠕动泵；3.限流阀；4.观察块；5.计数仪；6.显示器；7.观察池；8.细胞培养袋。In the figure, 1. Cell circulation tube; 1.1. Straight tube; 1.2. Elbow; 1.3. Connection head; 2. Peristaltic pump; 3. Restriction valve; 4. Observation block; 5. Counter; 6. Display; 7. . Observation pool; 8. Cell culture bag.
具体实施方式Detailed ways
以下结合附图和具体实施例对本申请作进一步的详细描述。The application will be further described in detail below in conjunction with the drawings and specific embodiments.
如图1～5所示的细胞计数装置，包括细胞循环管1、蠕动泵2、限流阀3、观察块4、计数仪5和显示器6；The cell counting device shown in Figures 1 to 5 includes a cell circulation tube 1, a peristaltic pump 2, a flow limiting valve 3, an observation block 4, a counter 5 and a display 6;
细胞循环管1设有至少两个用于与细胞培养袋8连通的开口处；The cell circulation tube 1 is provided with at least two openings for communicating with the cell culture bag 8;
观察块4为透明状，优选为无毒透明塑料材质，其内部设有与细胞循环管1连通的观察池7；观察池7包括入口端和出口端；观察池7可以为U形槽，也可以是其他形状。The observation block 4 is transparent, preferably a non-toxic transparent plastic material, and is provided with an observation pool 7 communicating with the cell circulation tube 1; the observation pool 7 includes an inlet end and an outlet end; the observation pool 7 can be a U-shaped groove, or Can be other shapes.
蠕动泵2与细胞循环管1连接；限流阀3与细胞循环管1连接并设于观察块4的入口端用于限定进入观察池7内的细胞液面厚度；观察池7内的细胞液面厚度为0～0.5mm，优选为0.1mm，这样可保证进入观察池7的细胞液内的细胞为单层，没有叠加，更利于计数。蠕动泵2和限流阀3可直接选用市场购买的运用于细胞培养系统上的相关型号产品。The peristaltic pump 2 is connected to the cell circulation tube 1; the restrictor valve 3 is connected to the cell circulation tube 1 and is arranged at the inlet end of the observation block 4 to limit the thickness of the cell liquid surface entering the observation pool 7; the cell liquid in the observation pool 7 The thickness of the surface is 0-0.5mm, preferably 0.1mm, which can ensure that the cells in the cell liquid entering the observation pool 7 are in a single layer without superimposition, which is more conducive to counting. The peristaltic pump 2 and the restrictor valve 3 can be directly selected from the relevant models purchased on the market and used in the cell culture system.
计数仪5包括纵向间隔设置的照明装置和显微镜，计数仪5与显示器6电连接；观察块4位于照明装置和显微镜之间。通过照明装置与显微镜(电子显微镜)的配合，可对观察池7内的细胞进行自动计数，并通过显示器6显示。当然，计数仪5还可以是其他现有技术，比如通过摄像头拍摄细胞图像进行计数的方式。本实施例的计数仪5实际上也是现有产品，因此具体的实施方式或结构不再进行叙述。使用时，将观察块4插入计数仪5内相应位置，打开蠕动泵2、限流阀3和显示器6即可。The counter 5 includes an illuminating device and a microscope arranged at intervals in the longitudinal direction, and the counter 5 is electrically connected to the display 6; the observation block 4 is located between the illuminating device and the microscope. Through the cooperation of the illumination device and the microscope (electron microscope), the cells in the observation cell 7 can be automatically counted and displayed on the display 6. Of course, the counter 5 can also be other existing technologies, such as a method of taking cell images through a camera for counting. The counter 5 of this embodiment is actually an existing product, so the specific implementation or structure will not be described again. When in use, insert the observation block 4 into the corresponding position in the counter 5, and turn on the peristaltic pump 2, the flow limiting valve 3 and the display 6.
细胞循环管1可以是一个U形管，管的两端即为开口处分别插入细胞培养袋8内，管的一部分为透明段，并设于限流阀3与蠕动泵2之间，因此只需要通过计数仪5对该段透明段进行拍摄计数即可。The cell circulation tube 1 can be a U-shaped tube. The two ends of the tube are the openings respectively inserted into the cell culture bag 8. A part of the tube is a transparent section and is set between the restrictor valve 3 and the peristaltic pump 2. It is necessary to use the counter 5 to photograph and count the transparent section.
本实施例的细胞循环管1采用另外一种形式，即细胞循环管1包括两根对称设置的管体，如图3、5所示，其中一个管体的一端 与观察池7的入口端连通，另一个管体的一端与观察池7的出口端连通，观察池7的入口端和出口端取决于蠕动泵2的方向。The cell circulation tube 1 of this embodiment adopts another form, that is, the cell circulation tube 1 includes two symmetrically arranged tube bodies, as shown in Figures 3 and 5, one end of which is connected to the inlet end of the observation pool 7 , One end of the other tube is connected to the outlet end of the observation tank 7, and the inlet and outlet ends of the observation tank 7 depend on the direction of the peristaltic pump 2.
管体可以包括多个直管1.1、弯头1.2和连接头1.3；各直管1.1和/或弯头1.2通过连接头1.3连接。通过直管1.1和弯头1.2的数量及配合方式可以调节管体的长度和布置形式；可以适用于不同尺寸的细胞培养袋8。The pipe body may include a plurality of straight pipes 1.1, elbows 1.2, and connectors 1.3; each straight pipe 1.1 and/or elbow 1.2 is connected by the connector 1.3. The length and arrangement of the tube body can be adjusted by the number and matching mode of the straight pipe 1.1 and the elbow 1.2; it can be applied to cell culture bags 8 of different sizes.
另外，为了降低细胞的损失，管体包括内管和外管；内管与观察池7连通。外管起到保护内管的作用。In addition, in order to reduce cell loss, the tube body includes an inner tube and an outer tube; the inner tube is in communication with the observation pool 7. The outer tube protects the inner tube.
如图3所示，直管1.1上设有多个通孔。通孔的设置可保证细胞培养袋8内不同位置的细胞都可进入观察池7内，保证了计数的准确性。虽然如图5所示的直管1.1上并没有设置通孔，但并不影响其正常工作。本实施例的细胞计数装置还可包括与细胞培养袋8配合的密封塞，以保证管体与细胞培养袋8连接的密封性。As shown in Figure 3, the straight pipe 1.1 is provided with multiple through holes. The arrangement of the through holes can ensure that cells at different positions in the cell culture bag 8 can enter the observation pool 7 to ensure the accuracy of counting. Although there is no through hole on the straight pipe 1.1 as shown in Fig. 5, it does not affect its normal operation. The cell counting device of this embodiment may further include a sealing plug matched with the cell culture bag 8 to ensure the tightness of the connection between the tube body and the cell culture bag 8.

Claims (8)

  1. 一种细胞计数装置，其特征在于：包括细胞循环管(1)、蠕动泵(2)、限流阀(3)、观察块(4)、计数仪(5)和显示器(6)；A cell counting device, which is characterized in that it comprises a cell circulation tube (1), a peristaltic pump (2), a restrictor valve (3), an observation block (4), a counter (5) and a display (6);
    所述细胞循环管(1)设有至少两个用于与细胞培养袋(8)连通的开口处；The cell circulation tube (1) is provided with at least two openings for communicating with the cell culture bag (8);
    所述观察块(4)为透明状，其内部设有与细胞循环管(1)连通的观察池(7)；所述观察池(7)包括入口端和出口端；The observation block (4) is transparent, and an observation pool (7) connected with the cell circulation tube (1) is arranged inside; the observation pool (7) includes an inlet end and an outlet end;
    所述蠕动泵(2)与细胞循环管(1)连接；所述限流阀(3)与细胞循环管(1)连接并设于观察块(4)的入口端用于限定进入观察池(7)内的细胞液面厚度；The peristaltic pump (2) is connected to the cell circulation tube (1); the restrictor valve (3) is connected to the cell circulation tube (1) and is arranged at the entrance end of the observation block (4) for restricting access to the observation pool ( 7) The thickness of the inner cell liquid surface;
    所述计数仪(5)包括纵向间隔设置的照明装置和显微镜，所述计数仪(5)与显示器(6)电连接；所述观察块(4)位于所述照明装置和显微镜之间。The counter (5) includes an illuminating device and a microscope arranged at intervals in the longitudinal direction, and the counter (5) is electrically connected with a display (6); the observation block (4) is located between the illuminating device and the microscope.
  2. 根据权利要求1所述的细胞计数装置，其特征在于：所述细胞循环管(1)包括两根对称设置的管体，其中一个管体的一端与观察池(7)的入口端连通，另一个管体的一端与观察池(7)的出口端连通。The cell counting device according to claim 1, characterized in that: the cell circulation tube (1) comprises two symmetrically arranged tube bodies, one end of the tube body is connected with the inlet end of the observation pool (7), and the other One end of a tube is connected with the outlet end of the observation tank (7).
  3. 根据权利要求2所述的细胞计数装置，其特征在于：所述管体包括多个直管(1.1)、弯头(1.2)和连接头(1.3)；各直管(1.1)和/或弯头(1.2)通过连接头(1.3)连接。The cell counting device according to claim 2, characterized in that: the tube body comprises a plurality of straight tubes (1.1), elbows (1.2) and connectors (1.3); each straight tube (1.1) and/or elbow The head (1.2) is connected via the connector (1.3).
  4. 根据权利要求2所述的细胞计数装置，其特征在于：所述管体包括内管和外管；所述内管与观察池(7)连通。The cell counting device according to claim 2, characterized in that: the tube body includes an inner tube and an outer tube; the inner tube is in communication with the observation pool (7).
  5. 根据权利要求3所述的细胞计数装置，其特征在于：所述直管(1.1)上设有多个通孔。The cell counting device according to claim 3, characterized in that: the straight pipe (1.1) is provided with a plurality of through holes.
  6. 根据权利要求1所述的细胞计数装置，其特征在于：所述观察池(7)内的细胞液面厚度为0～0.5mm。The cell counting device according to claim 1, characterized in that the thickness of the cell liquid surface in the observation pool (7) is 0-0.5 mm.
  7. 根据权利要求6所述的细胞计数装置，其特征在于：所述观察池(7)内的细胞液面厚度为0.1mm。The cell counting device according to claim 6, characterized in that: the thickness of the cell liquid surface in the observation pool (7) is 0.1 mm.
  8. 根据权利要求1所述的细胞计数装置，其特征在于：所述观察池(7)为U形槽。The cell counting device according to claim 1, wherein the observation pool (7) is a U-shaped groove.
PCT/CN2019/124300 2019-12-10 2019-12-10 Cell counting device WO2021114086A1 (en)

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