Micro-fluidic chip device for sperm climbing experiment and experiment method of micro-fluidic chip device

Abstract

The invention discloses a micro-fluidic chip device for a sperm climbing experiment and an experiment method thereof.The device comprises a micro-fluidic chip body and a temperature control device, a sperm sampling pool of the micro-fluidic chip body is filled with a human sperm sample, and the sperm sampling pool is communicated with a sperm enrichment pool through a sperm filtering channel; the sperm climbing channel is vertically fixed at the central position of the sperm enrichment pool through a lap joint block; a treating fluid sample inlet is formed in the top of the sperm climbing channel; a first sperm sampling opening, a second sperm sampling opening and a third sperm sampling opening are sequentially formed in the side surface of the sperm climbing channel; the temperature control device comprises a heater and a temperature controller, and the heater is arranged at the bottom of the sperm enrichment pool and used for maintaining the stability of the temperature microenvironment in the micro-fluidic chip main body; the microenvironment of the fallopian tube is accurately simulated through bionic design, and physiological support is provided for sperm activity evaluation.

Description

Technical Field

[0001] The present invention belongs to the technical field of assisted reproduction, and in particular relates to a microfluidic chip device for a sperm climbing experiment and an experimental method thereof. Background Art

[0002] As a key method for assessing sperm motility, the sperm climbing height test plays an important role in male fertility diagnosis. It is based on the autonomous motility of sperm in a specific culture medium. By observing the height to which sperm climb in a vertical tube, it objectively reflects their motility rate and activity.

[0003] However, traditional experimental methods have significant limitations, which restrict the effectiveness and accuracy of their clinical applications: 1. Efficiency bottleneck: relying on manual operation and control tests, the process is cumbersome and the throughput is low. The experiment needs to strictly control the time nodes and environmental conditions, takes up to several hours, and it is difficult to achieve rapid screening of large-scale samples. 2. Interference factors: Fluctuations in environmental temperature and humidity, and sample contamination risks (such as microbial contamination or chemical residues) can easily lead to deviations in results. For example, if semen is exposed to a non-constant temperature environment for more than 30 minutes, sperm motility may decrease by more than 40%. 3. Information limitations: only the static climbing distance is recorded, and there is a lack of dynamic analysis of sperm motion trajectory (such as movement speed, trajectory curvature, etc.), which is more correlated with the success rate of fertilization. Summary of the Invention

[0004] The present invention provides a microfluidic chip device and experimental method for sperm climbing experiments, which utilizes the advantages of high throughput, automation and precise control of microfluidic chips to solve the problems of complex operation, poor result repeatability and difficulty in simulating physiological environments in the existing technology.

[0005] To solve the above problems, the present invention provides the following technical solutions:

[0006] The embodiment of the present invention provides a device of a microfluidic chip for a sperm climbing experiment, comprising a microfluidic chip body and a temperature control device, wherein the microfluidic chip body comprises a substrate (1) and a sperm sampling pool (2), a sperm filtering channel (3), a sperm enrichment pool (4), and a sperm climbing channel (6) arranged on the substrate (1), wherein the sperm sampling pool (2) is filled with a human sperm sample, the sperm sampling pool (2) is connected to the sperm enrichment pool (4) through the sperm filtering channel (3), and the sperm climbing channel (6) is vertically fixed to the substrate (1) by a bridge block (10). At the center of the sperm enrichment pool (4), a treatment liquid sampling port (11) is provided on the top of the sperm climbing channel (6), and the treatment liquid is injected into the sperm climbing channel (6) through the treatment liquid sampling port (11). The treatment liquid slowly fills the sperm enrichment pool (4) through the sperm climbing channel (6), providing a fluid environment for subsequent sperm climbing; the temperature control device includes a heater (5) and a temperature controller, and the heater (5) is provided at the bottom (4-1) of the sperm enrichment pool (4) and is used to maintain the stability of the temperature microenvironment inside the microfluidic chip body;

[0007] A first sperm sampling port (7), a second sperm sampling port (8) and a third sperm sampling port (9) are sequentially arranged on the side of the sperm climbing channel (6); and the through holes of the first sperm sampling port (7), the second sperm sampling port (8) and the third sperm sampling port (9) are connected to the inner cavity of the sperm climbing channel (6).

[0008] In a preferred embodiment of the present invention, the first sperm sampling port (7), the second sperm sampling port (8) and the third sperm sampling port (9) are all annular boss structures with a diameter of 1-3 mm for efficiently injecting treatment fluid.

[0009] In a preferred embodiment of the present invention, the height between the first sperm sampling port (7) and the bottom of the sperm climbing channel (6) is 3 cm, the height between the second sperm sampling port (8) and the bottom of the sperm climbing channel (6) is 5 cm, and the height between the third sperm sampling port (9) and the bottom of the sperm climbing channel (6) is 8 cm, which are used to collect sperm of different climbing distances for dynamic analysis of movement trajectories.

[0010] In a preferred embodiment of the present invention, the substrate (1) is a circular cover glass or a glass slide with a diameter of 3-5 cm. The substrate (1) is used to overlap with the sperm sampling pool (2) and the sperm enrichment pool (4) to form a closed channel; the sperm sampling pool (2) is an annular structure for storing sperm samples, the inner diameter of the sperm sampling pool (2) is 1-3 mm, the outer diameter is 2-3 cm, and the height is 1-3 cm; the sperm filtering channel (3) is a grid structure with a grid size (3-1) of 10-100 μm, and is used to filter impurities in the sperm sample.

[0011] In a preferred embodiment of the present invention, the sperm enrichment pool (4) is a circular structure for collecting filtered sperm, the diameter of the sperm enrichment pool (4) is 2-3 cm, and the height thereof is 1-3 cm; the sperm climbing channel (6) is a cylindrical structure for completing a sperm climbing experiment, the inner diameter of the sperm climbing channel (6) is 1-3 mm, the outer diameter thereof is 3-5 mm, and the height thereof is 10-15 cm.

[0012] In a preferred embodiment of the present invention, the material of the microfluidic chip body is one or more materials selected from polydimethylsiloxane (PDMS), polycarbonate (PC), polymethyl methacrylate (PMMA) and pure glass with good biocompatibility; the material of the sperm filtration channel (3) is one or more materials selected from nylon, glass fiber and stainless steel with good chemical compatibility; the material of the heater (5) is ITO, nanosilver or PI heater with a diameter of 2-3 cm; the temperature controller is a digital temperature controller with a temperature adjustment range of 36.5-37.5°C, which is used to adjust the temperature of the heater.

[0013] The embodiment of the present invention provides an experimental method for a microfluidic chip device for a sperm climbing experiment, comprising the following steps:

[0014] Step 1, injecting a treatment liquid into the sperm climbing channel (6) through the treatment liquid inlet (11), the treatment liquid slowly filling the sperm enrichment pool (4) through the sperm climbing channel (6), and providing a fluid environment for the subsequent addition of sperm; the treatment liquid is selected from sperm culture medium, sperm capacitation fluid or human fallopian tube fluid;

[0015] Step 2: Start the temperature controller and activate the heater (5), and accurately adjust the temperature of the microfluidic chip body through the temperature control device to ensure that the internal environment is stably maintained within the preset temperature range, providing a temperature environment for the subsequent addition of sperm;

[0016] Step 3, adding the processed human sperm sample to the sperm sampling pool (2), effectively filtering out impurities and dead sperm in the semen through the sperm filtration channel (2), and achieving directional enrichment of active sperm. Subsequently, the active sperm in the sperm enrichment pool (4) are guided into the sperm climbing channel (6) to perform a sperm climbing experiment;

[0017] Step 4, collecting highly active sperms from the sperm sampling port of the sperm climbing channel (6), detecting and analyzing their motion trajectory parameters, including active sperm rate, sperm curve velocity, sperm linear velocity and sperm average velocity.

[0018] In a preferred embodiment of the present invention, the treatment fluid in step 1 is Vitrolife assisted reproductive IVF-sperm washing and insemination fluid, product model: G-IVF PLUS / 60mL-10136; step 2 specifically includes: encapsulating a heater (5) at the bottom of a sperm enrichment tank (4), arranging a sperm sampling tank (2), a sperm filtration channel (3), a sperm enrichment tank (4), a sperm climbing channel (6) and a treatment fluid inlet on a substrate (1), and forming a closed channel through a stacking process to obtain a microfluidic chip body.

[0019] An embodiment of the present invention provides an application of a microfluidic chip device for a sperm climbing experiment in the sperm climbing experiment.

[0020] Compared with the prior art, the embodiments of the present invention provide a microfluidic chip device and experimental method for sperm climbing experiments, which have the following beneficial effects: (1) The present invention accurately simulates the physiological environment: by simulating the fallopian tube microenvironment through bionic design, it provides physiological support for sperm activity assessment. (2) The present invention has efficient automated operability: it integrates inlet filtration, flow rate control and data acquisition functions, improves the repeatability of results, is easy to operate, is convenient for people in related fields to operate, and is more user-friendly. (3) The present invention has broad application prospects: it is easy to industrialize and can be used in clinical sperm function testing, assisted reproductive technology optimization, reproductive medicine research and other fields. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] In order to more clearly illustrate the technical solutions in the embodiments or the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are only some embodiments of the invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0022] Figure 1 A schematic structural diagram of a microfluidic chip device for sperm climbing experiments provided in an embodiment of the present application.

[0023] Figure 2A schematic cross-sectional view of a microfluidic chip device for sperm climbing experiments provided in an embodiment of the present application.

[0024] Figure 3 A schematic cross-sectional view of a sperm filtration channel provided in an embodiment of the present application.

[0025] Figure 4 A schematic diagram of the bottom structure of a sperm enrichment tank provided in an embodiment of the present application.

[0026] Figure 5 Analysis of human sperm motion trajectory during sperm sorting using the chip in Example 1 of the present invention; A: analysis of sperm motion trajectory in a 3 cm climbing experiment; B: analysis of sperm motion trajectory in a 5 cm climbing experiment; C: analysis of sperm motion trajectory in an 8 cm climbing experiment.

[0027] Figure 6 This is the human sperm quality analysis when the chip in Example 1 of the present invention is used for human sperm sorting; A: comparison of active sperm rate; B: comparison of sperm curve velocity; C: comparison of sperm linear velocity; D: comparison of sperm average velocity. DETAILED DESCRIPTION

[0028] The technical solutions in the embodiments of the present application will be described clearly and completely below in conjunction with the drawings in the embodiments of the present application. Obviously, the described embodiments are only a part of the embodiments of the present application, not all of the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative work are within the scope of protection of this application. The "upper", "lower", "front", "rear", "left", "right", etc. used in the installation position or direction of the structure or parts of this embodiment are based on the orientation of the given drawings. They are only for the convenience of expression to distinguish the relative positions of the various parts or directions, and do not represent the orientation of the device or functional parts of this embodiment when in use.

[0029] Microfluidics technology, through micron-scale channel design and precise fluid control, offers a novel solution for sperm function testing. Its advantages include: 1. Physiological environment simulation: It can construct biomimetic structures that mimic the fallopian tube microenvironment. For example, droplet microfluidics can simulate the fallopian tube's curvature and surface tension, recreating the mechanical conditions during sperm natural selection. 2. High-throughput automation: Multi-channel parallel testing increases sample processing efficiency by over 10 times. Combined with automated sample introduction and real-time imaging systems, it enables simultaneous analysis of multiple parameters, including sperm motility, morphology, and trajectory. 3. Dynamic data acquisition: The integration of high-speed photography and AI algorithms allows for tracking sperm's three-dimensional trajectory and accurately calculates key metrics such as average path velocity and linear velocity, providing a more comprehensive basis for clinical diagnosis. Therefore, the development of a microfluidics-based sperm climbing test chip not only overcomes the efficiency and accuracy limitations of traditional methods but also has the potential to transform male fertility assessment from static results to dynamic functions. For example, by simulating the fluid resistance and chemical gradients in different sections of the fallopian tube, sperm migration within the reproductive tract can be predicted, providing precise support for assisted reproductive technologies.

[0030] For this reason, Figure 1 and Figure 2 As shown, an embodiment of the present invention provides a device for a microfluidic chip for sperm climbing experiments, including a microfluidic chip body and a temperature control device. The microfluidic chip body includes a base 1 and a sperm sampling pool 2, a sperm filtration channel 3, a sperm enrichment pool 4, and a sperm climbing channel 6 disposed on the base 1. The sperm sampling pool 2 is filled with a human sperm sample and is connected to the sperm enrichment pool 4 via the sperm filtration channel 3. The sperm climbing channel 6 is vertically fixed to the center of the sperm enrichment pool 4 via a bridge block 10. A treatment liquid inlet 11 is provided at the top of the sperm climbing channel 6. The treatment liquid is injected into the sperm climbing channel 6 through the treatment liquid inlet 11. The treatment liquid slowly fills the sperm enrichment pool 4 through the sperm climbing channel 6, providing a fluid environment for subsequent sperm screening and culture. The temperature control device includes a heater 5 and a temperature controller. The heater 5 is disposed at the bottom 4-1 of the sperm enrichment pool 4 to maintain a stable temperature microenvironment within the microfluidic chip body. The treatment fluid of this embodiment can be selected from sperm culture medium, sperm capacitation fluid or human fallopian tube fluid. The treatment fluid is preferably Vitrolife Assisted Reproductive IVF-Sperm Washing and Insemination Fluid, product model: G-IVF PLUS / 60mL-10136.

[0031] The side of the sperm climbing channel 6 is provided with a first sperm sampling port 7, a second sperm sampling port 8 and a third sperm sampling port 9; and the through holes of the first sperm sampling port 7, the second sperm sampling port 8 and the third sperm sampling port 9 are connected to the inner cavity of the sperm climbing channel 6. The first treatment liquid inlet 6, the second treatment liquid inlet 7 and the third treatment liquid inlet 8 are all annular boss structures with a diameter of 1-3mm, which are used for efficient injection of treatment liquid. Preferably, the height between the first sperm sampling port 7 and the bottom of the sperm climbing channel 6 is 3cm high, the height between the second sperm sampling port 8 and the bottom of the sperm climbing channel 6 is 5cm high, and the height between the third sperm sampling port 9 and the bottom of the sperm climbing channel 6 is 8cm high, which are used to collect sperm of different climbing distances for dynamic analysis of movement trajectories.

[0032] Base 1 is a circular cover glass or slide with a diameter of 3-5 cm. It is used to overlap with sperm sampling reservoir 2 and sperm enrichment reservoir 4 to form a closed channel. Sperm sampling reservoir 2 is a ring-shaped structure used to store sperm samples. Its inner diameter is 1-3 mm, its outer diameter is 2-3 cm, and its height is 1-3 cm. Figure 3 Combine Figure 1 The sperm filtering channel 3 is a grid structure with a grid size 3-1 of 10-100 μm, which is used to filter impurities in the sperm sample. Figure 3 This is the cross-sectional structure of the sperm filtering channel 3, and its mesh size 3-1 is 10-100 μm.

[0033] The sperm collection tank 4 is a circular structure used to collect filtered sperm. The diameter of the sperm collection tank 4 is 2-3 cm and the height is 1-3 cm. The sperm climbing channel 6 is a cylindrical structure used to complete the sperm climbing experiment. The inner diameter of the sperm climbing channel 6 is 1-3 mm, the outer diameter is 3-5 mm, and the height is 10-15 cm.

[0034] The material of the microfluidic chip body is one or more of polydimethylsiloxane (PDMS), polycarbonate (PC), polymethyl methacrylate (PMMA) and pure glass with good biocompatibility; the material of the sperm filtration channel 3 is one or more of nylon, glass fiber and stainless steel with good chemical compatibility. Figure 4 Combine Figure 2 The heater 5 is made of ITO, nanosilver, or PI and has a diameter of 2-3 cm. The temperature controller is a digital thermostat with an adjustable temperature range of 36.5-37.5°C. It is used to adjust the temperature of the heater to maintain the temperature of the main area of the microfluidic chip within a certain range.

[0035] The embodiment of the present invention provides an experimental method for a microfluidic chip device for a sperm climbing experiment, comprising the following steps:

[0036] Step 1: injecting a treatment liquid into the sperm climbing channel 6 through the treatment liquid inlet 11. The treatment liquid slowly fills the sperm enrichment pool 4 through the sperm climbing channel 6 to provide a fluid environment for the subsequent addition of sperm; the treatment liquid is selected from sperm culture medium, sperm capacitation fluid or human fallopian tube fluid;

[0037] Step 2: Start the temperature controller and activate the heater 5 to precisely adjust the temperature of the microfluidic chip body through the temperature control device to ensure that the internal environment is stably maintained within the preset temperature range, providing a temperature environment for the subsequent addition of sperm;

[0038] Step 3: Add the processed human sperm sample to the sperm sampling pool 2, and effectively filter out impurities and dead sperm in the semen through the sperm filtration channel 2 to achieve directional enrichment of active sperm. Subsequently, the active sperm in the sperm enrichment pool 4 are guided into the sperm climbing channel 6 for a sperm climbing experiment;

[0039] Step 4: collect highly active sperm from the sperm sampling port of the sperm climbing channel 6, and detect and analyze their motion trajectory parameters, including active sperm rate, sperm curve velocity, sperm linear velocity and sperm average velocity.

[0040] The treatment liquid in step 1 is Vitrolife assisted reproductive IVF-sperm washing and insemination liquid, product model: G-IVF PLUS / 60mL-10136; step 2 specifically includes: encapsulating the heater 5 at the bottom of the sperm enrichment tank 4, and arranging the sperm sampling tank 2, sperm filtration channel 3, sperm enrichment tank 4, sperm climbing channel 6 and treatment liquid inlet on the base 1, and forming a closed channel through a superposition process to obtain the microfluidic chip body.

[0041] The preparation process of the microfluidic chip body in step 2 is as follows: the expected micropattern is drawn according to the CAD software, and then a mask is made according to the micropattern; the micropattern is developed on a silicon wafer to obtain a silicon wafer mold of the micropattern; using the organic material polydimethylsiloxane, unsolidified polydimethylsiloxane is poured on the silicon wafer mold, and it is heat-baked at 80°C for 1 hour to solidify, thereby obtaining an integrated area with several microchannels; the microchannel structure is cut and punched with a punch, and then the microchannel structure is bonded to the substrate after surface treatment using a plasma cleaner.

[0042] An embodiment of the present invention also provides a method for preparing a microfluidic chip device for sperm climbing experiments, the method comprising: obtaining the microfluidic chip body by using soft lithography technology or microinjection molding method; arranging the sperm sampling pool, sperm filtration channel, sperm enrichment pool, sperm climbing channel and treatment liquid inlet on the substrate, and forming a closed channel through a superposition process to obtain the microfluidic chip body.

[0043] An embodiment of the present invention further provides an application of the microfluidic chip device for sperm climbing experiment as described in the above embodiment in the sperm climbing experiment.

[0044] The following will describe in detail a microfluidic chip for sperm climbing experiment and its preparation method and application in conjunction with the accompanying drawings.

[0045] Example 1: A microfluidic chip for sperm climbing experiment and its preparation method

[0046] 1. A microfluidic chip device for sperm climbing experiment Figure 1 and Figure 2 As shown, it includes: a microfluidic chip body, including a substrate 1 and a sperm sampling pool 2, a sperm filtration channel 3, a sperm enrichment pool 4, a sperm climbing channel 6 and a treatment liquid inlet 11 arranged on the substrate 1, the bottom of the sperm sampling pool 2 is connected to the sperm enrichment pool 4 through the sperm filtration channel 3, and the sperm enrichment pool 4 is connected to the treatment liquid inlet 11 through the sperm climbing channel 6. A first sperm sampling port 7, a second sperm sampling port 8 and a third sperm sampling port 9 are arranged on the side of the sperm climbing channel 6. A temperature control device includes a heater 5 and a temperature controller. The heater 5 is arranged at the bottom 4-1 of the sperm enrichment pool 4, and is used to maintain the stability of the temperature microenvironment inside the microfluidic chip body, reference Figure 4 .

[0047] Base 1 is a 4 cm diameter coverslip, model Fisher-12-545-C. Sperm sampling reservoir 2 has an inner diameter of 2 mm, an outer diameter of 2.5 cm, and a height of 2 cm. Sperm filtration channel 3 has a pore size of 50 μm and is made of nylon. Sperm enrichment reservoir 4 has an inner diameter of 2.5 cm and a height of 2 cm. Sperm climbing channel 6 has an inner diameter of 2 mm, an outer diameter of 4 mm, and a height of 12.5 cm. The heights of the first, second, and third sperm sampling ports 7, 8, and 9 are 3, 5, and 8 cm, respectively. Heater 5 has a diameter of 2.5 cm and is made of ITO. The temperature controller is a digital OMRON-E5CC-800 thermostat, set at 37°C.

[0048] 2. The preparation method of the microfluidic chip for the sperm climbing experiment is as follows:

[0049] Step S1: Draw the desired micro pattern using CAD software, and then make a mask based on the micro pattern;

[0050] Step S2: developing the micro pattern on the silicon wafer by ultraviolet lithography to obtain a silicon wafer mold of the micro pattern;

[0051] Step S3: using an organic material, polydimethylsiloxane, to pour unsolidified polydimethylsiloxane onto a silicon wafer mold, and then heat-bake at 80° C. for 1 hour to solidify the mold, thereby obtaining a semi-finished product with a plurality of microchannels;

[0052] Step S4: encapsulate the heater at the bottom of the sperm enrichment tank, cut the microchannel structure and punch holes with a puncher, and then bond the microchannel structure to the substrate after surface treatment with a plasma cleaning machine.

[0053] Experimental Example 1: Human sperm sorting

[0054] The chips of the embodiment and the comparative example were used to sort human sperm, and the sperm sorting effect of each group was statistically analyzed. The sperm sorting steps included:

[0055] Step T1: 1 mL of human oviduct fluid is injected into the treatment fluid inlet, and sequentially passes through the sperm climbing channel and the sperm enrichment tank until the entire sperm enrichment tank is filled. Then, the treated 1 mL of human sperm sample is injected into the sperm injection tank;

[0056] Step T2: Start the temperature controller and activate the heater. The temperature control system accurately adjusts the temperature of the entire device to ensure that the internal environment is stably maintained at around 37°C, allowing the sperm in the sperm enrichment pool after filtration to swim into the sperm climbing channel.

[0057] Step T3: 30 minutes later, 5 μL of sperm samples were collected from the sperm sampling ports at heights of 3 cm, 5 cm, and 8 cm in the sperm climbing channel for sperm activity testing.

[0058] The sperm motility of the above examples was statistically analyzed, as shown in Table 1.

[0059] Table 1

[0060]

[0061] As shown in Table 1, in Example 1 of the present invention, there are significant differences in the active sperm rate, sperm curve velocity, sperm linear velocity and sperm average velocity of sperm sampled from the sperm climbing channel at heights of 3 cm, 5 cm and 8 cm. Figure 5 For the analysis of human sperm movement trajectory in Example 1 of the present invention, Figure 6 This is the sperm activity analysis in Example 1 of the present invention.

[0062] Finally, it should be noted that the terms "comprises," "includes," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that includes a list of elements includes not only those elements, but also other elements not expressly listed, or elements inherent to such process, method, article, or apparatus.

[0063] Although the preferred embodiments of the present invention have been described, those skilled in the art may make additional changes and modifications to these embodiments once they have learned the basic creative concept. Therefore, the appended claims are intended to be interpreted as including the preferred embodiments and all changes and modifications that fall within the scope of the present invention.

[0064] Obviously, those skilled in the art may make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if such changes and modifications fall within the scope of the claims and their equivalents, the present invention is intended to include such changes and modifications.

Claims

1. A microfluidic chip device for sperm climbing experiment, characterized in that: The invention comprises a microfluidic chip body and a temperature control device, wherein the microfluidic chip body comprises a substrate (1) and a sperm sampling pool (2), a sperm filtering channel (3), a sperm enrichment pool (4), and a sperm climbing channel (6) arranged on the substrate (1); the sperm sampling pool (2) is filled with a human sperm sample; the sperm sampling pool (2) is connected to the sperm enrichment pool (4) through the sperm filtering channel (3); and the sperm climbing channel (6) is vertically fixed to the center of the sperm enrichment pool (4) through a bridge block (10). The sperm climbing channel (6) is provided with a treatment liquid sampling port (11) at the top thereof, and the treatment liquid is injected into the sperm climbing channel (6) through the treatment liquid sampling port (11), and the treatment liquid slowly fills the sperm enrichment pool (4) through the sperm climbing channel (6), providing a fluid environment for subsequent sperm climbing; the temperature control device comprises a heater (5) and a temperature controller, and the heater (5) is provided at the bottom (4-1) of the sperm enrichment pool (4) and is used to maintain the stability of the temperature microenvironment inside the microfluidic chip body; A first sperm sampling port (7), a second sperm sampling port (8) and a third sperm sampling port (9) are sequentially arranged on the side of the sperm climbing channel (6); and the through holes of the first sperm sampling port (7), the second sperm sampling port (8) and the third sperm sampling port (9) are connected to the inner cavity of the sperm climbing channel (6).

2. The microfluidic chip device for sperm climbing experiment according to claim 1, characterized in that: The first sperm sampling port (7), the second sperm sampling port (8) and the third sperm sampling port (9) are all circular boss structures with a diameter of 1-3 mm, and are used for efficiently injecting treatment fluid.

3. The microfluidic chip device for sperm climbing experiment according to claim 2, characterized in that: The height between the first sperm sampling port (7) and the bottom of the sperm climbing channel (6) is 3 cm, the height between the second sperm sampling port (8) and the bottom of the sperm climbing channel (6) is 5 cm, and the height between the third sperm sampling port (9) and the bottom of the sperm climbing channel (6) is 8 cm, which are used to collect sperm with different climbing distances for dynamic analysis of movement trajectories.

4. The microfluidic chip device for sperm climbing experiment according to claim 1, characterized in that: The substrate (1) is a circular cover glass or a glass slide with a diameter of 3-5 cm. The substrate (1) is used to overlap with a sperm sampling pool (2) and a sperm enrichment pool (4) to form a closed channel. The sperm sampling pool (2) is an annular structure for storing sperm samples. The inner diameter of the sperm sampling pool (2) is 1-3 mm, the outer diameter is 2-3 cm, and the height is 1-3 cm. The sperm filtering channel (3) is a grid structure with a grid size (3-1) of 10-100 μm, and is used to filter impurities in the sperm sample.

5. The microfluidic chip device for sperm climbing experiment according to claim 1, characterized in that: The sperm enrichment pool (4) is a circular structure for collecting filtered sperm, the diameter of the sperm enrichment pool (4) is 2-3 cm, and the height thereof is 1-3 cm; the sperm climbing channel (6) is a cylindrical structure for completing a sperm climbing experiment, the inner diameter of the sperm climbing channel (6) is 1-3 mm, the outer diameter thereof is 3-5 mm, and the height thereof is 10-15 cm.

6. The microfluidic chip device for sperm climbing experiment according to claim 1, characterized in that: The material of the microfluidic chip body is one or more materials selected from polydimethylsiloxane (PDMS), polycarbonate (PC), polymethyl methacrylate (PMMA) and pure glass with good biocompatibility; the material of the sperm filtration channel (3) is one or more materials selected from nylon, glass fiber and stainless steel with good chemical compatibility; the material of the heater (5) is ITO, nanosilver or PI heater with a diameter of 2-3 cm; the temperature controller is a digital temperature controller with a temperature adjustment range of 36.5-37.5°C, which is used to adjust the temperature of the heater.

7. An experimental method for a microfluidic chip device for sperm climbing experiment, characterized in that: The following steps are involved: Step 1, injecting a treatment liquid into the sperm climbing channel (6) through the treatment liquid inlet (11), the treatment liquid slowly filling the sperm enrichment pool (4) through the sperm climbing channel (6), and providing a fluid environment for the subsequent addition of sperm; the treatment liquid is selected from sperm culture medium, sperm capacitation fluid or human fallopian tube fluid; Step 2: Start the temperature controller and activate the heater (5), and accurately adjust the temperature of the microfluidic chip body through the temperature control device to ensure that the internal environment is stably maintained within the preset temperature range, providing a temperature environment for the subsequent addition of sperm; Step 3, adding the processed human sperm sample to the sperm sampling pool (2), effectively filtering out impurities and dead sperm in the semen through the sperm filtration channel (2), and achieving directional enrichment of active sperm. Subsequently, the active sperm in the sperm enrichment pool (4) are guided into the sperm climbing channel (6) to perform a sperm climbing experiment; Step 4, collecting highly active sperms from the sperm sampling port of the sperm climbing channel (6), detecting and analyzing their motion trajectory parameters, including active sperm rate, sperm curve velocity, sperm linear velocity and sperm average velocity.

8. The experimental method of a microfluidic chip device for sperm climbing experiment according to claim 7, characterized in that: The treatment liquid in step 1 is Vitrolife assisted reproductive IVF-sperm washing and insemination liquid, product model: G-IVFPLUS / 60mL-10136; step 2 specifically includes: encapsulating the heater (5) at the bottom of the sperm enrichment tank (4), arranging the sperm sampling tank (2), the sperm filtration channel (3), the sperm enrichment tank (4), the sperm climbing channel (6) and the treatment liquid inlet on the base (1), and forming a closed channel through a stacking process to obtain the microfluidic chip body.

9. Use of the microfluidic chip device for sperm climbing height experiment as claimed in any one of claims 1 to 6 in a sperm climbing height experiment.

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