Micro-fluidic chip structure for oocyte separation and collection

Abstract

The utility model discloses a micro-fluidic chip structure for oocyte separation and collection, which comprises a shell, a first separation bin and a second separation bin are respectively arranged at two ends in the shell, and the first separation bin is communicated with the second separation bin; the first separation bin is externally communicated with an inlet, and the second separation bin is externally communicated with a recovery port; a first channel is arranged in the first separation bin, and first capturing pits are arrayed at the bottom of the first channel; the second separation bin internally comprises a second channel, and channel columns are arranged in the second channel in an array mode; according to the utility model, a culture solution can be directly injected into the shell from the inlet and can be recycled for repeated separation, so that the content of obtained cells is high, the separation is convenient and rapid, and the manpower is greatly saved.

Description

Technical Field

[0001] The utility model belongs to the technical field of medical experimental equipment, and relates to a microfluidic chip structure, in particular to a microfluidic chip structure used for separating and collecting oocytes. Background Art

[0002] In the experiment of studying mouse oocytes, it is necessary to separate oocytes from mouse ovaries. The separation method is as follows: transfer the cleaned mouse ovaries to a clean disposable culture dish, use a disposable sterile blade to fully mince the ovarian tissue into a viscous state, then use M2 culture medium containing 50μM I BMX to disperse the ovarian fragments, and gently shake the culture dish to fully release the oocytes;

[0003] However, the oocytes released by the above method are of different sizes. Researchers need to manually select oocytes of appropriate size and good morphology in the GV stage with a mouth pipette under a stereoscope. The diameter of these cells is between 70-90μm. The selected GV stage oocytes are then transferred to MI 16 culture medium and rinsed repeatedly several times to remove excess impurities. This method requires manual selection of oocytes, which not only has high technical requirements for technicians, but is also very time-consuming.

[0004] Patent CN207468631 U discloses a cell separation sieve, including a rectangular support frame, a frame, a first sieve and a second sieve; the cost of the cell sieve is reduced, and the experimental efficiency is improved; the granulosa cells around the oocyte are extracted, and the oocyte and granulosa cells are separated at the same time; however, the diameter of the sieve in this scheme is too large, and it is not suitable for the separation and collection of mouse oocytes;

[0005] Therefore, there is no microfluidic chip structure for oocyte separation and collection that can quickly separate and collect oocytes with a diameter of 70-90 μm to improve the work efficiency of researchers. Utility Model Content

[0006] The utility model aims to provide a microfluidic chip structure for separating and collecting oocytes, so as to solve the problem of how to quickly separate and collect oocytes with a diameter of 70-90 μm.

[0007] The purpose of the utility model can be achieved through the following technical solutions:

[0008] A microfluidic chip structure for oocyte separation and collection, comprising: a shell, a first separation chamber and a second separation chamber;

[0009] A first separation chamber and a second separation chamber are respectively arranged at two ends inside the shell, and the first separation chamber and the second separation chamber are communicated with each other;

[0010] An entrance is arranged outside the first separation chamber, and a recovery port is arranged outside the second separation chamber;

[0011] The first separation chamber includes a first channel, and a first capture pit is arranged in an array at the bottom of the first channel, and the diameter of the first capture pit is 70 μm;

[0012] The second separation chamber includes a second channel, and channel columns are arranged in an array in the second channel. The spacing between the channel columns is 90 μm.

[0013] Furthermore, the depth of the first capture pit is 100 μm.

[0014] Furthermore, the inlet is communicated with the first channel, and the recovery port is communicated with the second channel.

[0015] Furthermore, the second separation chamber also includes a recovery liquid injection port and an outlet.

[0016] Furthermore, the second separation chamber is connected to the first separation chamber through a pipeline.

[0017] Furthermore, the recovery liquid injection port is connected to the second channel, and the recovery liquid injection port and the recovery port are symmetrically arranged at two ends of the second separation chamber.

[0018] Furthermore, the outlet and the inlet are symmetrically arranged at two ends of the shell.

[0019] Furthermore, a second capture pit is arranged in an array at the bottom of the second channel, and the diameter of the second capture pit is 90 μm.

[0020] Furthermore, the depth of the second capture pit is 100 μm.

[0021] Beneficial effects of the utility model:

[0022] In the utility model, a first separation chamber is provided to separate cells and impurities with a cell diameter less than 70 μm in the culture medium, and then a second separation chamber is provided to separate cells with a diameter greater than 90 μm, thereby obtaining cells with a target diameter of 70-90 μm. The utility model can directly inject the culture medium into the shell from the inlet, and the culture medium can be recovered for repeated separation, the obtained cell content is high, the separation is convenient and fast, and manpower is greatly saved. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] The utility model is further described below in conjunction with the accompanying drawings.

[0024] Figure 1 It is a schematic diagram of one of the overall structures of the utility model;

[0025] Figure 2 yes Figure 1 Schematic diagram of the internal structure;

[0026] Figure 3 It is a schematic diagram of the second overall structure of the utility model;

[0027] Figure 4 yes Figure 3 Schematic diagram of the internal structure;

[0028] In the figure: 1. Shell; 2. Inlet; 3. Recovery port; 4. Outlet; 5. Recovery injection port; 6. First separation chamber; 601. First channel; 602. First capture pit; 7. Second separation chamber; 701. Second channel; 702. Channel column; 703. Second capture pit. DETAILED DESCRIPTION

[0029] The following will be combined with the drawings in the embodiments of the utility model to clearly and completely describe the technical solutions in the embodiments of the utility model. Obviously, the described embodiments are only part of the embodiments of the utility model, not all of the embodiments. Based on the embodiments of the utility model, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the utility model.

[0030] Example 1

[0031] See also Figure 1-Figure 2 As shown, the utility model is a microfluidic chip structure for oocyte separation and collection, comprising: a housing 1, a first separation chamber 6 and a second separation chamber 7;

[0032] A first separation chamber 6 is provided at one end of the shell 1, and a second separation chamber 7 is provided at the other end of the shell 1, and the first separation chamber 6 and the second separation chamber 7 are connected; an inlet 2 is provided outside the first separation chamber 6, and a recovery port 3 is provided outside the second separation chamber 7;

[0033] The first separation chamber 6 is composed of a first channel 601 and a first capture pit 602 at the bottom of the first channel 601. The first capture pit 602 array is arranged at the bottom of the first channel 601. The diameter of the first capture pit 602 is 70 μm, and the depth of the first capture pit 602 is 100 μm.

[0034] The second separation chamber 7 is composed of a second channel 701 and a channel column 702 in the second channel 701. The channel column 702 array is arranged in the second channel 701. The spacing between the channel columns 702 is 90 μm, which facilitates the passage of cells in the culture solution while limiting the passage of cells with a diameter of more than 90 μm.

[0035] The method of using the utility model is:

[0036] Step 1: Load the culture solution containing oocytes from the inlet 2 into the first separation chamber 6. The culture solution is naturally settled in the first channel 601 in the first separation chamber 6, so that cells and impurities with a diameter less than 70 μm in the culture solution are settled into the first capture pit 602. After the sedimentation is completed, the oocytes larger than 70 μm are in the first channel 601 above the capture pit;

[0037] Step 2: Inject MI 16 culture medium into the first separation chamber 6 from the inlet 2 to push the cells into the second separation chamber 7 from the first channel 601 in the first separation chamber 6, so that the cells with a diameter less than 90 μm in the culture medium pass through the channel columns 702 with a spacing of 90 μm and enter the second channel 701 of the second separation chamber 7. At this time, the second channel 701 contains oocytes with a cell diameter of 70-90 μm, and then the oocytes are collected from the recovery port 3 into the culture dish to complete the separation and collection of the cells.

[0038] Example 2

[0039] See also Figure 3-Figure 4 As shown, the utility model is a microfluidic chip structure for oocyte separation and collection, comprising: a housing 1, a first separation chamber 6 and a second separation chamber 7;

[0040] A first separation chamber 6 is provided at one end of the shell 1, and a second separation chamber 7 is provided at the other end of the shell 1, and the first separation chamber 6 and the second separation chamber 7 are connected; an inlet 2 is provided outside the first separation chamber 6, and a recovery port 3 is provided outside the second separation chamber 7;

[0041] One end of the first separation chamber 6 and one end of the second separation chamber 7 in the housing 1 are connected through a pipeline;

[0042] The other end of the first separation chamber 6 is connected to the inlet 2, the other end of the second separation chamber 7 is connected to the outlet 4, and the upper and lower ends of the second separation chamber 7 are respectively provided with a recovery port 3 and a recovery liquid injection port 5;

[0043] The first separation chamber 6 is composed of a first channel 601 and a first capture pit 602 at the bottom of the first channel 601. The first capture pit 602 array is arranged at the bottom of the first channel 601. The diameter of the first capture pit 602 is 70 μm, and the depth of the first capture pit 602 is 100 μm.

[0044] The second separation chamber 7 is composed of a second channel 701 and a second capture pit 703 at the bottom of the second channel 701. The second capture pit 703 array is arranged at the bottom of the second channel 701. The diameter of the second capture pit 703 is 90 μm, and the depth of the second capture pit 703 is 100 μm.

[0045] The method of using the utility model is:

[0046] Step 1: Load the culture solution containing oocytes from the inlet 2 into the first separation chamber 6. The culture solution is naturally settled in the first channel 601 in the first separation chamber 6, so that cells and impurities with a diameter less than 70 μm in the culture solution are settled into the first capture pit 602. After the sedimentation is completed, the oocytes larger than 70 μm are in the first channel 601 above the capture pit;

[0047] Step 2: Inject MI 16 culture medium into the first separation chamber 6 from the inlet 2 to push the cells from the first channel 601 in the first separation chamber 6 into the second channel 701 in the second separation chamber 7, so that the cells with a diameter of 70-90 μm in the culture medium are precipitated into the second capture pit 703. After the sedimentation is completed, the cells larger than 90 μm in the second channel 701 are discharged from the outlet 4;

[0048] Step three, open the recovery injection port 5 and the recovery port 3, close the inlet 2 and the outlet 4, invert the shell 1, allow the 70-90 μm oocytes to settle back into the channel from the second capture pit 703, and then inject MI 16 culture medium from the recovery injection port 5, so that the oocytes are collected from the recovery port 3 into the culture dish, completing the separation and collection of the cells.

[0049] The above is a detailed description of an embodiment of the utility model, but the content is only a preferred embodiment of the utility model and cannot be considered to limit the scope of implementation of the utility model. All equivalent changes and improvements made within the scope of application of the utility model should still fall within the scope of the patent coverage of the utility model.

Claims

1. A microfluidic chip structure for oocyte separation and collection, characterized in that: include: A housing (1), a first separation chamber (6) and a second separation chamber (7); The first separation chamber (6) and the second separation chamber (7) are respectively arranged at two ends inside the shell (1), and the first separation chamber (6) and the second separation chamber (7) are communicated with each other; An inlet (2) is arranged outside the first separation chamber (6), and a recovery port (3) is arranged outside the second separation chamber (7); The first separation chamber (6) includes a first channel (601), and a first capture pit (602) is arranged in an array at the bottom of the first channel (601), and the diameter of the first capture pit (602) is 70 μm; The second separation chamber (7) comprises a second channel (701), and channel columns (702) are arranged in an array in the second channel (701), and the spacing between the channel columns (702) is 90 μm.

2. A microfluidic chip structure for oocyte separation and collection according to claim 1, characterized in that: The depth of the first capture pit (602) is 100 μm.

3. A microfluidic chip structure for oocyte separation and collection according to claim 1, characterized in that: The inlet (2) is in communication with the first channel (601), and the recovery port (3) is in communication with the second channel (701).

4. A microfluidic chip structure for oocyte separation and collection according to claim 1, characterized in that: The second separation chamber (7) further comprises a liquid recovery inlet (5) and an outlet (4).

5. A microfluidic chip structure for oocyte separation and collection according to claim 4, characterized in that: The second separation bin (7) and the first separation bin (6) are connected via a pipeline.

6. A microfluidic chip structure for oocyte separation and collection according to claim 4, characterized in that: The recovery liquid injection port (5) is connected to the second channel (701), and the recovery liquid injection port (5) and the recovery port (3) are symmetrically arranged at two ends of the second separation chamber (7).

7. A microfluidic chip structure for oocyte separation and collection according to claim 4, characterized in that: The outlet (4) and the inlet (2) are symmetrically arranged at two ends of the shell (1).

8. A microfluidic chip structure for oocyte separation and collection according to claim 6, characterized in that: The second channel (701) is provided with a second capture pit (703) in an array at the bottom thereof, and the diameter of the second capture pit (703) is 90 μm.

9. A microfluidic chip structure for oocyte separation and collection according to claim 8, characterized in that: The depth of the second capture pit (703) is 100 μm.

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