Sperm sorting micro-fluidic chip based on ultrasonic waves as well as preparation method and application of sperm sorting micro-fluidic chip

Abstract

The invention discloses a body ultrasonic wave-based sperm sorting micro-fluidic chip and a preparation method and application thereof, the sperm sorting micro-fluidic chip comprises an integrated micro-fluidic chip main body and a body ultrasonic wave device, the integrated micro-fluidic chip main body comprises a substrate and an integrated region structure arranged on the substrate, the integration area structure comprises a sperm sampling pool, a sperm sorting channel and a sperm enrichment pool, and the bottom of the sperm sampling pool is communicated with the bottom of the sperm enrichment pool through the sperm sorting channel; the body ultrasonic device comprises an arbitrary waveform function generator and ultrasonic transducers, and the ultrasonic transducers are arranged on the two sides of the sperm sorting channel and used for generating a standing wave sound field; sperms swim to the sperm enrichment pool from the sperm sampling pool through the sperm sorting channel in the ultrasonic standing wave field, dead sperms and sperms with poor activity cannot swim into the sperm enrichment pool under the action of the sound field, and high-activity sperms can swim into the sperm enrichment pool, so that the active sperms can be sorted.

Description

Technical Field

[0001] The present invention belongs to the technical field of assisted reproduction, and in particular relates to a sperm sorting microfluidic chip based on body ultrasound, and a preparation method and application thereof. Background Art

[0002] Sperm sorting is of great significance in assisted reproduction and prevention of genetic diseases, especially in in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI). Selecting healthy and motile sperm is crucial to improving the success rate of pregnancy and the birth rate of healthy babies. Existing sperm sorting technologies mainly rely on traditional methods such as density gradient centrifugation and motility screening, but these methods have disadvantages such as complex operation, low sorting efficiency, and potential damage to sperm. In addition, traditional sperm sorting technology is difficult to perform high-throughput, non-destructive and efficient sperm sorting at the microscale.

[0003] The emergence of microfluidics technology has provided new possibilities for sperm sorting. Microfluidic chips can perform fine manipulation at the micrometer scale and achieve directional manipulation of biological particles through fluid mechanics and physical fields (such as electric fields, magnetic fields, and acoustic fields). However, most of the current sperm sorting methods based on microfluidics have problems such as low sorting efficiency and limited sperm concentration.

[0004] Therefore, it is necessary to develop an efficient and non-destructive microfluidic sperm sorting method based on bulk ultrasound. Summary of the invention

[0005] The present invention provides a sperm sorting microfluidic chip based on body ultrasound, and a preparation method and application thereof. The non-contact and high efficiency of body ultrasound are utilized to achieve accurate sperm sorting in the microfluidic chip, solving the problems of low sorting efficiency and severe sperm damage in the prior art.

[0006] To solve the above problems, the technical solution provided by the present invention is as follows:

[0007] The embodiment of the present invention provides a sperm sorting microfluidic chip based on bulk ultrasound, comprising an integrated microfluidic chip body (1) and a bulk ultrasound device (2), wherein the integrated microfluidic chip body (1) comprises a substrate (10) and an integrated area structure (20) arranged on the substrate (10), wherein the integrated area structure (20) comprises a sperm injection pool (201), a sperm sorting channel (202) and a sperm enrichment pool (203), wherein the bottom of the sperm injection pool (201) is connected to the bottom of the sperm enrichment pool (203) through the sperm sorting channel (202);

[0008] The body ultrasonic wave device (2) comprises an arbitrary waveform function generator (21) and an ultrasonic transducer (22). The ultrasonic transducer (22) is arranged on both sides of the sperm sorting channel (202) and is used to generate a standing wave acoustic field, so that sperm can swim from the sperm injection pool (201) through the sperm sorting channel (202) to the sperm enrichment pool (203) in the body ultrasonic wave standing wave field. Under the action of the acoustic field, dead sperm and sperm with poor vitality cannot swim into the sperm enrichment pool (203), while highly active sperm swim into the sperm enrichment pool (203), thereby achieving the sorting of active sperm.

[0009] In an optional embodiment of the present invention, the substrate (10) is a cover glass or a slide glass, which is used to overlap with the integrated area structure (20) to form a closed channel. The length of the substrate (10) is 2 to 10 cm and its width is 1 to 3 cm.

[0010] In an optional embodiment of the present invention, the sperm injection pool (201) is a circular structure for storing sperm samples, and a sperm injection hole is provided in the middle of the top thereof. The diameter of the sperm injection pool (201) is 2 to 10 mm, and the height is 1 to 10 mm.

[0011] In an optional embodiment of the present invention, the sperm sorting channel (202) is used to sort sperm with higher activity, and has a width of 0.5 to 5 mm, a length of 5 to 25 mm, and a height of 50 to 500 μm.

[0012] In an optional embodiment of the present invention, the sperm enrichment pool (203) is a circular structure for collecting sorted sperm, and a sperm enrichment hole is provided in the middle of the top thereof. The diameter of the sperm enrichment pool (203) is 2 to 10 mm and the height is 1 to 10 mm.

[0013] In an optional embodiment of the present invention, the ultrasonic transducer (22) is a rectangular piezoelectric ceramic piece with a length of 5 to 25 mm, a width of 1 to 5 mm, a thickness of 1 to 5 mm, and a resonance frequency of 1 to 10 MHz.

[0014] In an optional embodiment of the present invention, the arbitrary waveform function generator (21) has an output frequency of 1 to 10 MHz and an amplitude output of 1 to 10 Vpp.

[0015] The present invention provides a method for preparing a sperm sorting microfluidic chip based on body ultrasound as in the above embodiment, comprising the following steps:

[0016] Step 1, obtaining an integrated area structure (20) by using soft lithography technology or micro injection molding method, wherein the integrated area structure (20) includes a sperm injection pool (201), a sperm sorting channel (202) and a sperm enrichment pool (203);

[0017] Step 2, placing the integrated area structure (20) on a substrate (10), with the bottom of the integrated area structure (20) overlapping with the substrate (10) to form a closed channel, thereby obtaining an integrated microfluidic chip body (1);

[0018] Step 3, providing a body ultrasonic wave device (2), the body ultrasonic wave device (2) comprising an arbitrary waveform function generator (21) and an ultrasonic transducer (22) electrically connected to each other, the ultrasonic transducer (22) being arranged on both sides of the sperm sorting channel (202) to generate a standing wave acoustic field, so that sperm can swim from the sperm injection pool (201) through the sperm sorting channel (202) to the sperm enrichment pool (203) in the body ultrasonic wave standing wave field, and under the action of the acoustic field, dead sperm and sperm with poor motility cannot swim into the sperm enrichment pool (203), while highly active sperm can swim into the sperm enrichment pool (203), thereby achieving the sorting of active sperm.

[0019] The present invention also provides an application of a sperm sorting microfluidic chip based on body ultrasound as in the above embodiment in sperm sorting.

[0020] Compared with the prior art, the embodiments of the present invention provide a sperm sorting microfluidic chip based on body ultrasound and a preparation method and application thereof, which have the following beneficial effects:

[0021] (1) The present invention can achieve efficient sorting of active sperm through precise control of body ultrasound.

[0022] (2) The non-contact manipulation of the body ultrasound of the present invention avoids mechanical damage to sperm and can achieve non-destructive sorting of active sperm.

[0023] (3) In the sperm sorting process, the present invention does not require filtration and centrifugation, is easy to operate, and is convenient for personnel in related fields to operate. It is more user-friendly and easy to industrialize, and has great application prospects and commercial value. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] In order to more clearly illustrate the technical solutions in the embodiments or the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. 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.

[0025] Figure 1 A schematic diagram of a sperm sorting microfluidic chip based on body ultrasound provided in an embodiment of the present application.

[0026] Figure 2Schematic diagram of the analysis of the movement trajectory of unsorted sperm when the sperm sorting microfluidic chip provided in an embodiment of the present application performs sperm sorting.

[0027] Figure 3 Schematic diagram of sperm movement trajectory analysis by non-body ultrasonic sorting when the sperm sorting microfluidic chip provided in an embodiment of the present application is used for sperm sorting.

[0028] Figure 4 Schematic diagram of sperm movement trajectory analysis when the sperm sorting microfluidic chip provided in an embodiment of the present application is used for sperm sorting by bulk ultrasound sorting.

[0029] Figure 5 A schematic diagram of human sperm quality analysis - comparison of active sperm rates when the chip provided in an embodiment of the present application is used for human sperm sorting.

[0030] Figure 6 Schematic diagram of human sperm quality analysis - sperm curve rate comparison when the chip provided in the embodiment of the present application is used for human sperm sorting.

[0031] Figure 7 A schematic diagram of human sperm quality analysis - sperm linear velocity comparison when the chip provided in an embodiment of the present application is used for human sperm sorting.

[0032] Figure 8 A schematic diagram of human sperm quality analysis - sperm average rate comparison when the chip provided in an embodiment of the present application is used for human sperm sorting. DETAILED DESCRIPTION

[0033] 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 part of the embodiments of the present application, rather than 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 the present application.

[0034] Throughout the specification, unless otherwise specifically stated, the terms used herein should be understood as meanings commonly used in the art. Therefore, unless otherwise defined, all technical and scientific terms used herein have the same meanings as those generally understood by technicians in the field to which the present invention belongs. If there is a conflict, this specification takes precedence. It should be noted that when an element is referred to as being fixed to or "set on" another element, it can be directly on another element or indirectly on another element; when an element is referred to as being "connected to" another element, it can be directly connected to another element or indirectly connected to another element. It should be understood that the orientation or positional relationship indicated by the terms "length", "width", "upper", "lower", "front", "back", "first", "second", "vertical", "horizontal", "top", "bottom", "inside", "outside", etc. is based on the orientation or positional relationship shown in the drawings, which is only for the convenience of describing the present application and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as a limitation to the present application. In addition, in the description of the present application, the meaning of "multiple" and "several" is two or more, unless otherwise clearly and specifically defined.

[0035] like Figure 1 As shown, an embodiment of the present invention provides a sperm sorting microfluidic chip based on bulk ultrasound, including an integrated microfluidic chip body 1 and a bulk ultrasound device 2. The integrated microfluidic chip body 1 includes a substrate 10 and an integrated area structure 20 arranged on the substrate 10. The integrated area structure 20 includes a sperm injection pool 201, a sperm sorting channel 202 and a sperm enrichment pool 203. The bottom of the sperm injection pool 201 is connected to the bottom of the sperm enrichment pool 203 through the sperm sorting channel 202.

[0036] The body ultrasonic wave device 2 includes an arbitrary waveform function generator 21 and an ultrasonic transducer 22. The ultrasonic transducer 22 is arranged on both sides of the sperm sorting channel 202 to generate a standing wave sound field so that sperm can swim from the sperm injection pool 201 through the sperm sorting channel 202 to the sperm enrichment pool 203 in the body ultrasonic wave standing wave field. Under the action of the sound field, dead sperm and sperm with poor vitality cannot swim into the sperm enrichment pool 203, while highly active sperm can swim into the sperm enrichment pool 203, thereby realizing the sorting of active sperm.

[0037] The substrate 10 is a cover glass or a slide glass, which is used to overlap with the integrated area structure 20 to form a closed channel. The length of the substrate 10 is 2 to 10 cm, and its width is 1 to 3 cm. The sperm sampling pool 201 is a circular structure, which is used to store sperm samples. The middle part of its top is provided with a sperm sampling hole. The diameter of the sperm sampling pool 201 is 2 to 10 mm, and its height is 1 to 10 mm. The sperm sorting channel 202 is used to sort sperm with higher activity. Its width is 0.5 to 5 mm, its length is 5 to 25 mm, and its height is 50 to 500 μm. The sperm enrichment pool 203 is a circular structure, which is used to collect sorted sperm. The middle part of its top is provided with a sperm enrichment hole. The diameter of the sperm enrichment pool 203 is 2 to 10 mm, and its height is 1 to 10 mm.

[0038] The ultrasonic transducer 22 is a rectangular piezoelectric ceramic piece with a length of 5-25 mm, a width of 1-5 mm, a thickness of 1-5 mm, and a resonance frequency of 1-10 MHz. The arbitrary waveform function generator 21 has an output frequency of 1-10 MHz and an amplitude output of 1-10 Vpp.

[0039] The material of the above-mentioned integrated microfluidic chip body 1 is polydimethylsiloxane (PDMS) with good biocompatibility, and other microfluidic chip materials, such as PC, PMMA, pure glass and other materials, can also be used. The arbitrary waveform function generator 21 is used to provide an excitation signal to the ultrasonic transducer, and can adjust the frequency, amplitude and phase of the signal within a certain range. The ultrasonic transducer 22 converts the high-frequency electrical signal into a corresponding high-frequency acoustic signal through the piezoelectric effect, which is used to generate a sound field of a specific frequency. The two poles of the ultrasonic transducer 22 are led out through wires and are electrically connected to the arbitrary waveform function generator 21 respectively.

[0040] A sperm injection hole is provided in the middle of the top of the sperm injection pool 201 of this embodiment. The shape of the sperm injection pool 201 can be circular, square, rectangular or any other shape. The height of the sperm injection pool 201 is 1 to 10 mm, and the height of the sperm injection hole is 0.1 to 1 mm. A sperm enrichment hole is provided in the middle of the top of the sperm enrichment pool 203. The shape of the sperm enrichment pool 203 can be circular, square, rectangular or any other shape. The height of the sperm enrichment pool 203 is 1 to 10 mm, and the height of the sperm enrichment hole is 0.1 to 1 mm. The width of the sperm sorting channel 202 is 0.5 to 5 mm, the length is 5 to 25 mm, and the height is 50 to 500 μm. The sperm sorting effect within the length, width and height range of the sperm sorting channel 202 is better.

[0041] The present invention also provides a method for preparing a sperm sorting microfluidic chip based on body ultrasound as in the above embodiment, characterized in that it comprises the following steps:

[0042] Step 1, using soft lithography technology or micro injection molding method to obtain an integrated area structure 20, wherein the integrated area structure 20 includes a sperm injection pool 201, a sperm sorting channel 202 and a sperm enrichment pool 203; specifically, drawing the expected micro pattern according to CAD software, and then making a mask according to the micro pattern; developing the micro pattern on a silicon wafer to obtain a silicon wafer mold of the micro pattern; using an organic material polydimethylsiloxane, pouring unsolidified polydimethylsiloxane on the silicon wafer mold, and heat baking at 80°C for 1 hour to solidify, to obtain an integrated area structure with a plurality of microchannels.

[0043] Step 2, the integrated area structure 20 is arranged on the substrate 10, and the bottom of the integrated area structure 20 is overlapped with the substrate 10 to form a closed channel, so as to obtain an integrated microfluidic chip body 1. Specifically, the integrated area structure 20 of the microchannel is cut and punched with a puncher, and then the microchannel structure is bonded to the substrate 10 after surface treatment by a plasma cleaning machine.

[0044] Step 3, providing a body ultrasonic wave device 2, the body ultrasonic wave device 2 includes an arbitrary waveform function generator 21 and an ultrasonic transducer 22 electrically connected to each other, the ultrasonic transducer 22 is arranged on both sides of the sperm sorting channel 202, and generates a standing wave sound field, so that sperm can swim from the sperm injection pool 201 through the sperm sorting channel 202 to the sperm enrichment pool 203 in the body ultrasonic wave standing wave field, and under the action of the sound field, dead sperm and sperm with poor vitality cannot swim into the sperm enrichment pool 203, and highly active sperm can swim into the sperm enrichment pool 203, thereby realizing the sorting of active sperm.

[0045] The sperm sorting process of the sperm sorting microfluidic chip based on body ultrasound is as follows: first, the treatment liquid is added to the sperm sampling pool, and the treatment liquid passes through the sperm sorting channel and the sperm enrichment pool in turn. At this time, the treatment liquid fills the sperm sorting channel to provide the required environment for the subsequent addition of sperm. Then, the treated human sperm sample is added to the sperm sampling pool, and then the microfluidic chip is placed in an incubator and the arbitrary waveform function generator is turned on to allow the sperm to swim to the sperm enrichment pool through the sperm sorting channel in the body ultrasound standing wave field. Finally, the highly active sperm in the sperm enrichment pool is collected, and the dead sperm and sperm with poor vitality cannot swim into the sperm enrichment pool under the action of the acoustic field, thereby realizing the sorting of active sperm. Among them, the treatment liquid can be selected from sperm culture fluid, sperm capacitation fluid or human fallopian tube fluid. The treatment liquid of the embodiment of the present invention is Vitrolife assisted reproductive IVF-washing sperm insemination fluid, product model: G-IVF PLUS / 60mL-10136.

[0046] The embodiment of the present invention also provides an application of a sperm sorting microfluidic chip based on body ultrasound as described in the above embodiment in sperm sorting.

[0047] The separation effects of Example 1 of the present invention and Comparative Example 2 are compared, and the specific operations are as follows:

[0048] A sperm sorting microfluidic chip based on bulk ultrasound in Example 1 of the present invention comprises an integrated microfluidic chip body 1 and a bulk ultrasound device 2. The integrated microfluidic chip body 1 comprises a substrate 10, an integrated area structure 20 arranged on the substrate, the integrated area structure 20 comprises a sperm injection pool 201, a sperm sorting channel 202, a sperm enrichment pool 203, and an ultrasonic transducer 204, the bottom of the sperm injection pool 201 is connected to the bottom of the sperm enrichment pool 203 through the sperm sorting channel 202. The bulk ultrasound device 2 comprises an arbitrary waveform function generator 21 and an ultrasonic transducer 22, the ultrasonic transducer 22 is arranged on both sides of the sperm sorting channel 202, and is used to generate a standing wave acoustic field.

[0049] The substrate 10 is a 22×40mm cover glass, and its model is Fisher-12-545-C. The sperm sampling pool 201 has a diameter of 6mm and a height of 6mm, and the sperm sampling hole has a diameter of 0.4mm and a height of 0.4mm. The sperm sorting channel 202 has a width of 0.8mm, a length of 15mm, and a height of 100μm. The sperm enrichment pool 203 has a diameter of 4mm and a height of 6mm; the sperm enrichment hole has a diameter of 0.2mm and a height of 0.4mm. The ultrasonic transducer 22 is a rectangular piezoelectric ceramic piece with a length of 15mm, a width of 5mm, a thickness of 1mm, and a resonant frequency of 2MHz. The model of the arbitrary waveform function generator 21 is AFG3000, which has a dual-channel design, a maximum sampling rate of 1GS / s, an output frequency of up to 100MHz, and a maximum amplitude output of 10Vpp. In this embodiment, the arbitrary waveform function generator outputs a 2 MHz sine wave signal with an amplitude of 10 Vpp and a phase of 0.

[0050] The method for preparing a sperm sorting microfluidic chip based on body ultrasound in Example 1 of the present invention is as follows:

[0051] Step 1: Draw the expected micro pattern according to the CAD software, and then make a mask according to the micro pattern; develop the micro pattern on the silicon wafer through ultraviolet lithography technology to obtain a silicon wafer mold with the micro pattern.

[0052] Step 2: using organic material polydimethylsiloxane, pouring unsolidified polydimethylsiloxane on the silicon wafer mold, and baking it at 80° C. for 1 hour to solidify it, thereby obtaining a semi-finished product with a plurality of microchannels.

[0053] Step 3, encapsulate the ultrasonic transducer on both sides of the sperm sorting channel, 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.

[0054] In Comparative Example 2, no body ultrasound was applied during the sperm sorting process, and other structures and steps were similar to those of Example 1 of the present invention.

[0055] The chips of Example 1 of the present invention and Comparative Example 2 were used to sort human sperm, and the sperm sorting effect of each group was counted, wherein the sperm sorting step includes:

[0056] Step 1: Inject 1 mL of human fallopian tube fluid into the sperm injection pool, and pass it through the sperm sorting channel and sperm enrichment pool in sequence until the entire sperm sorting channel is filled, and then inject the processed 200 μL human sperm sample into the sperm injection hole.

[0057] Step 2, place the entire device in an incubator at 37°C and a carbon dioxide concentration of 5%, turn on the arbitrary waveform function generator, and allow sperm to swim from the sperm sampling pool through the sperm sorting channel in the body ultrasonic standing wave field to the sperm enrichment pool.

[0058] Step 3: 20 minutes later, take 5 μL of sorted sperm sample from the sperm enrichment pool for sperm activity detection.

[0059] The sorting effects of Example 1 of the present invention and Comparative Example 2 are statistically shown in Table 1.

[0060] Table 1

[0061]

[0062] It can be seen from Table 1 that in Comparative Example 2, the active sperm rate, sperm curve velocity, sperm linear velocity and sperm average velocity of sperm obtained by non-body ultrasonic sorting are lower. In Example 1 of the present invention, the active sperm rate, sperm curve velocity, sperm linear velocity and sperm average velocity of sperm obtained by body ultrasonic sorting are higher.

[0063] Figure 2 , Figure 3 , Figure 4 For the analysis of human sperm movement trajectory in Example 1 of the present invention, Figure 5 , Figure 6 , Figure 7 and Figure 8 This is the sperm activity analysis in Example 1 of the present invention. It can be seen that the sperm activity obtained by the in vivo ultrasonic wave sorting of the present invention is higher.

[0064] 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 series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements that are inherent to such process, method, article, or apparatus.

[0065] 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.

[0066] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. A sperm sorting microfluidic chip based on body ultrasound, characterized in that: The invention comprises an integrated microfluidic chip body (1) and a body ultrasonic wave device (2), wherein the integrated microfluidic chip body (1) comprises a substrate (10) and an integrated area structure (20) arranged on the substrate (10), wherein the integrated area structure (20) comprises a sperm injection pool (201), a sperm sorting channel (202) and a sperm enrichment pool (203), and the bottom of the sperm injection pool (201) is connected to the bottom of the sperm enrichment pool (203) through the sperm sorting channel (202); The body ultrasonic wave device (2) comprises an arbitrary waveform function generator (21) and an ultrasonic transducer (22). The ultrasonic transducer (22) is arranged on both sides of the sperm sorting channel (202) and is used to generate a standing wave acoustic field, so that sperm can swim from the sperm injection pool (201) through the sperm sorting channel (202) to the sperm enrichment pool (203) in the body ultrasonic wave standing wave field. Under the action of the acoustic field, dead sperm and sperm with poor vitality cannot swim into the sperm enrichment pool (203), while highly active sperm can swim into the sperm enrichment pool (203), thereby achieving the sorting of active sperm.

2. The sperm sorting microfluidic chip based on body ultrasound according to claim 1, characterized in that: The substrate (10) is a cover glass or a slide glass, and is used to overlap with the integrated area structure (20) to form a closed channel. The length of the substrate (10) is 2 to 10 cm, and the width is 1 to 3 cm.

3. The sperm sorting microfluidic chip based on body ultrasound according to claim 1, characterized in that: The sperm sampling pool (201) is a circular structure for storing sperm samples, and a sperm sampling hole is provided in the middle of the top thereof. The diameter of the sperm sampling pool (201) is 2 to 10 mm, and the height is 1 to 10 mm.

4. The sperm sorting microfluidic chip based on body ultrasound according to claim 1, characterized in that: The sperm sorting channel (202) is used to sort sperm with higher activity, and has a width of 0.5 to 5 mm, a length of 5 to 25 mm, and a height of 50 to 500 μm.

5. The sperm sorting microfluidic chip based on body ultrasound according to claim 1, characterized in that: The sperm enrichment pool (203) is a circular structure, used for collecting sorted sperm, and a sperm enrichment hole is provided in the middle of the top thereof. The sperm enrichment pool (203) has a diameter of 2 to 10 mm and a height of 1 to 10 mm.

6. The sperm sorting microfluidic chip based on body ultrasound according to claim 1, characterized in that: The ultrasonic transducer (22) is a rectangular piezoelectric ceramic piece with a length of 5 to 25 mm, a width of 1 to 5 mm, a thickness of 1 to 5 mm, and a resonance frequency of 1 to 10 MHz.

7. The sperm sorting microfluidic chip based on body ultrasound according to claim 1, characterized in that: The arbitrary waveform function generator (21) has an output frequency of 1 to 10 MHz and an amplitude output of 1 to 10 Vpp.

8. A method for preparing a sperm sorting microfluidic chip based on bulk ultrasound according to any one of claims 1 to 7, characterized in that: The following steps are involved: Step 1, obtaining an integrated area structure (20) by using soft lithography technology or micro injection molding method, wherein the integrated area structure (20) includes a sperm injection pool (201), a sperm sorting channel (202) and a sperm enrichment pool (203); Step 2, placing the integrated area structure (20) on a substrate (10), with the bottom of the integrated area structure (20) overlapping with the substrate (10) to form a closed channel, thereby obtaining an integrated microfluidic chip body (1); Step 3, providing a body ultrasonic wave device (2), the body ultrasonic wave device (2) comprising an arbitrary waveform function generator (21) and an ultrasonic transducer (22) electrically connected to each other, the ultrasonic transducer (22) being arranged on both sides of the sperm sorting channel (202) to generate a standing wave acoustic field, so that sperm can swim from the sperm injection pool (201) through the sperm sorting channel (202) to the sperm enrichment pool (203) in the body ultrasonic wave standing wave field, and under the action of the acoustic field, dead sperm and sperm with poor motility cannot swim into the sperm enrichment pool (203), while highly active sperm can swim into the sperm enrichment pool (203), thereby achieving the sorting of active sperm.

9. Use of the sperm sorting microfluidic chip based on bulk ultrasound according to any one of claims 1 to 7 in sperm sorting.

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