Sperm collection method for reproductive medicine laboratory

Abstract

The invention relates to the technical field of sperm collection, and discloses a sperm collection method for a reproductive medicine laboratory, which comprises the following steps: cleaning and operating an experimental mouse to accurately collect sperms of the experimental mouse; the method comprises the following steps: firstly, cleaning the abdomen of an experimental mouse and performing surgical incision to expose testis and epididymis; then, separating the epididymis from the testis by using a sterilized appliance, and flushing the epididymis with normal saline by using an injector; sperms are extracted from the flushing fluid, and distribution of the sperms in liquid drops is obtained through computer simulation and image analysis; according to the method, accurate image acquisition and simulation calculation are combined, sperm distribution can be accurately judged, the suction strategy is optimized, and it is ensured that the maximum number of sperms are collected; finally, the sperms are separated through centrifugal operation; the method provides a new accurate scheme for sperm collection, and has high reliability and operability.

Description

Technical Field

[0001] The present invention relates to the technical field of sperm collection, and specifically provides a sperm collection method for reproductive medicine laboratories. Background Art

[0002] In reproductive medicine research and clinical practice, sperm collection is a crucial step that directly affects the quality of subsequent experiments and the accuracy of results. Traditional sperm collection methods usually rely on manual operations and simple mechanical assistance. However, various challenges are faced during sperm collection. Firstly, sperm collection is usually carried out by flushing the epididymis. However, it is impossible to accurately evaluate the distribution and quantity of sperm. This method often relies on manual observation, with low precision, and it is impossible to avoid sperm loss caused by improper operation or environmental factors. In addition, traditional methods lack detailed monitoring and optimization strategies for sperm distribution. During the operation, the distribution of sperm droplets is highly non-uniform. How to accurately determine which areas have a concentrated sperm population and effectively aspirate these sperm remains a bottleneck that is difficult to break through in current technologies. Many existing technologies lack an in-depth understanding of fluid mechanics during sperm collection. Sperm in the droplets will change their positions under the influence of fluid mechanics during aspiration, and traditional collection methods cannot accurately predict and control these dynamic changes.

[0003] In summary, there is an urgent need for a sperm collection method for reproductive medicine laboratories to accurately evaluate the sperm distribution in the droplets to be aspirated, optimize the aspiration strategy, maximize the sperm collection efficiency, and significantly improve the precision and effectiveness of sperm collection. Summary of the Invention

[0004] The present invention provides a sperm collection method for reproductive medicine laboratories, which helps to solve the problems mentioned in the above background art.

[0005] The present invention provides the following technical solution: A sperm collection method for reproductive medicine laboratories, comprising:

[0006] Preparing experimental mice for sperm collection;

[0007] Cleaning the abdomen of the experimental mice, specifically:

[0008] Using a cotton ball soaked with 75% alcohol to evenly wipe the abdomen of the experimental mice;

[0009] After cleaning, using a sterilized scissors to cut open the abdominal cavity of the experimental mice;

[0010] Pulling the peritesticular fat to separate the testis and epididymis outside the experimental mice;

[0011] Preparing two sterilized culture dishes, denoted as the first culture dish and the second culture dish respectively;

[0012] Place the testis and epididymis into the first culture dish;

[0013] Separate the epididymis from the testis in the first culture dish, and place the separated epididymis into the second culture dish;

[0014] In the second culture dish, rinse the epididymis according to the rinsing strategy, specifically:

[0015] Use a syringe to aspirate 5 ml of 0.9% physiological saline that has been in a 37°C water bath;

[0016] Insert the needle of the syringe into an epididymis, and slowly push out the physiological saline in the syringe so that the pushed-out physiological saline forms a droplet. Place the rinsed epididymis into a sterilized centrifuge tube;

[0017] Perform the same rinsing operation on the other epididymis of the experimental mouse so that the physiological saline pushed out in the two rinses forms a droplet;

[0018] Place the two rinsed epididymides into the same sterilized centrifuge tube;

[0019] The sterilized centrifuge tube containing the two rinsed epididymides is denoted as the target centrifuge tube;

[0020] Denote the droplet formed by the physiological saline pushed out in the two rinses as the droplet to be aspirated;

[0021] Obtain the distribution of sperm in the droplet to be aspirated;

[0022] Aspirate the sperm in the droplet to be aspirated according to the aspiration strategy;

[0023] Transfer the aspirated sperm to the target centrifuge tube;

[0024] Centrifuge the target centrifuge tube to collect sperm.

[0025] Optionally, the obtaining the distribution of sperm in the droplet to be aspirated specifically includes:

[0026] Slowly move the second culture dish to the sample stage of the optical coherence tomography device, and keep the relative position between the second culture dish and the droplet to be aspirated in it unchanged during the movement;

[0027] Use a low-coherence light beam to scan the droplet to be aspirated layer by layer, and obtain the reflection signals at different positions of the droplet to be aspirated;

[0028] According to the reflection signals at different positions in the droplet to be aspirated, convert the reflection signal of each layer into an image signal to form a set of tomographic images from light to deep;

[0029] Each layer of the image corresponds to a cross-section at a different position within the droplet;

[0030] Perform image processing on each layer of the acquired images, enhance the low-contrast regions, and improve the contrast between the sperm and the surrounding background;

[0031] Synthesize each layer of the image after image processing into a three-dimensional space, denoted as the three-dimensional image of the droplet to be aspirated, and display the distribution of sperm in the three-dimensional image of the droplet to be aspirated.

[0032] Optionally, obtaining the distribution of sperm in the droplet to be aspirated according to the sperm distribution acquisition strategy specifically includes:

[0033] Obtain the three-dimensional image of the droplet to be aspirated, and display the distribution of sperm in the three-dimensional image of the droplet to be aspirated;

[0034] Uniformly distribute points in the three-dimensional image of the droplet to be aspirated, specifically:

[0035] Use a number of planes parallel to the horizontal plane to divide the three-dimensional image of the droplet to be aspirated, and the distance between each plane is 0.1 mm. The number of planes parallel to the horizontal plane constitutes the first plane set;

[0036] Use a number of planes perpendicular to the planes in the first plane set to divide the three-dimensional image of the droplet to be aspirated, and the distance between each plane is 0.1 mm. The number of planes perpendicular to the planes in the first plane set constitutes the second plane set;

[0037] Use a number of planes perpendicular to the planes in both the first plane set and the second plane set to divide the three-dimensional image of the droplet to be aspirated, and the distance between each plane is 0.1 mm. The number of planes perpendicular to the planes in both the first plane set and the second plane set constitutes the third plane set;

[0038] The planes in the first plane set, the planes in the second plane set, and the planes in the third plane set intersect to form a number of intersection points;

[0039] Obtain the intersection points located inside the three-dimensional image of the droplet to be aspirated, and denote them as the first test point, the second test point... the Nth test point, where N represents the number of intersection points located inside the three-dimensional image of the droplet to be aspirated.

[0040] Optionally, aspirating sperm in the droplet to be aspirated according to the aspiration strategy further includes:

[0041] Set the single aspiration volume;

[0042] The single aspiration volume refers to the volume aspirated at one time when aspirating sperm in the droplet to be aspirated, and the single aspiration volume is equal to 1 ml;

[0043] Obtain the thickness of the liquid droplet to be aspirated, denoted as the standard thickness;

[0044] Calculate the single aspiration volume divided by the standard thickness, and record the result as the maximum aspiration area;

[0045] In the three-dimensional image of the liquid droplet to be aspirated, draw a circle with the first test point as the center and an area equal to the maximum aspiration area, denoted as the first circular surface, and the first circular surface is parallel to the horizontal plane;

[0046] Obtain the number of sperm within the first circular surface, denoted as the first quantity.

[0047] Optionally, the method of aspirating sperm from the liquid droplet to be aspirated according to the aspiration strategy further includes:

[0048] When the aspiration point is the first test point and the aspiration volume is the single aspiration volume, simulate the flow process of the liquid inside the liquid droplet to be aspirated through computer, specifically including:

[0049] Before the start of aspiration, mark the liquid at the second test point, the third test point... the Nth test point, and record the positions of the liquid at the second test point, the third test point... the Nth test point at this time;

[0050] Obtain the moment when aspiration starts, denoted as the start moment;

[0051] Starting from the start moment, every time 0.1 ml is aspirated, obtain the real-time positions of the liquid at the second test point, the third test point... the Nth test point until the aspiration is completed;

[0052] Model the shape of the remaining liquid droplet after aspiration and the sperm distributed therein according to the real-time positions of the liquid at the second test point, the third test point... the Nth test point during the aspiration process, denoted as the three-dimensional image of the first auxiliary liquid droplet;

[0053] Uniformly distribute points in the three-dimensional image of the first auxiliary liquid droplet, specifically:

[0054] Make a number of planes parallel to the horizontal plane to divide the three-dimensional image of the first auxiliary liquid droplet, and the distance between each plane is 0.1 mm. The number of planes parallel to the horizontal plane constitutes the first auxiliary plane set;

[0055] Make a number of planes perpendicular to the planes in the first auxiliary plane set to divide the three-dimensional image of the first auxiliary liquid droplet, and the distance between each plane is 0.1 mm. The number of planes perpendicular to the planes in the first auxiliary plane set constitutes the second auxiliary plane set;

[0056] Make several planes that are simultaneously perpendicular to the planes in the first auxiliary plane set and the second auxiliary plane set to divide the three-dimensional image of the first auxiliary droplet. The distance between each plane is 0.1 mm. The several planes that are simultaneously perpendicular to the planes in the first auxiliary plane set and the second auxiliary plane set form a third auxiliary plane set;

[0057] The planes in the first auxiliary plane set, the planes in the second auxiliary plane set, and the planes in the third auxiliary plane set intersect to form several auxiliary intersection points;

[0058] In the three-dimensional image of the first auxiliary droplet, draw a circle with an area equal to the maximum aspiration area centered on each auxiliary intersection point. Each circular surface is parallel to the horizontal plane;

[0059] And respectively obtain the number of sperm in each circular surface;

[0060] Obtain the circular surface with the largest number of sperm, denoted as the first auxiliary circular surface;

[0061] Obtain the number of sperm in the first auxiliary circular surface, denoted as the first auxiliary quantity;

[0062] The auxiliary intersection point corresponding to the center of the first auxiliary circular surface is denoted as the first auxiliary point;

[0063] Calculate the first quantity + the first auxiliary quantity, and record the result as the first simulated quantity.

[0064] Optionally, the method of aspirating sperm from the droplet to be aspirated according to the aspiration strategy further includes:

[0065] In the three-dimensional image of the droplet to be aspirated, draw a circle with an area equal to the maximum aspiration area centered on the second test point, denoted as the second circular surface, and the second circular surface is parallel to the horizontal plane;

[0066] Obtain the number of sperm in the second circular surface, denoted as the second quantity;

[0067] When the aspiration point is the second test point and the aspiration volume is the single aspiration volume, use a computer to simulate the liquid flow process inside the droplet to be aspirated, specifically including:

[0068] Before the aspiration starts, mark the liquid at the first test point, the third test point... the Nth test point, and record the positions of the liquid at the first test point, the third test point... the Nth test point at this time;

[0069] Obtain the start time of the aspiration, denoted as the start time;

[0070] Starting from the start time, obtain the real-time positions of the liquid at the first test point, the third test point... the Nth test point every 0.1 ml of aspiration until the aspiration ends;

[0071] Model the shape of the remaining droplets and the sperm distributed therein after aspiration based on the real-time positions of the liquid at the first test point, the third test point,..., the Nth test point during the aspiration process, and record it as the three-dimensional image of the second auxiliary droplet;

[0072] Uniformly distribute points in the three-dimensional image of the second auxiliary droplet, specifically:

[0073] Make a number of planes parallel to the horizontal plane to divide the three-dimensional image of the second auxiliary droplet, and the distance between each plane is 0.1 mm. The number of planes parallel to the horizontal plane constitutes the fourth auxiliary plane set;

[0074] Make a number of planes perpendicular to the planes in the fourth auxiliary plane set to divide the three-dimensional image of the second auxiliary droplet, and the distance between each plane is 0.1 mm. The number of planes perpendicular to the planes in the fourth auxiliary plane set constitutes the fifth auxiliary plane set;

[0075] Make a number of planes perpendicular to the planes in both the fourth auxiliary plane set and the fifth auxiliary plane set to divide the three-dimensional image of the second auxiliary droplet, and the distance between each plane is 0.1 mm. The number of planes perpendicular to the planes in both the fourth auxiliary plane set and the fifth auxiliary plane set constitutes the sixth auxiliary plane set;

[0076] The planes in the fourth auxiliary plane set, the planes in the fifth auxiliary plane set, and the planes in the sixth auxiliary plane set intersect to form a number of auxiliary intersection points;

[0077] In the three-dimensional image of the second auxiliary droplet, draw a circle with an area equal to the maximum aspiration area with each auxiliary intersection point as the center, and each circular surface is parallel to the horizontal plane;

[0078] And respectively obtain the number of sperm in each circular surface;

[0079] Obtain the circular surface with the largest number of sperm, and record it as the second auxiliary circular surface;

[0080] Obtain the number of sperm in the second auxiliary circular surface, and record it as the second auxiliary quantity;

[0081] The auxiliary intersection point corresponding to the center of the first auxiliary circular surface is recorded as the first auxiliary point;

[0082] Calculate the second quantity + the second auxiliary quantity, and record the result as the second simulated quantity;

[0083] Through the above method, respectively obtain the third quantity, the fourth quantity,..., the Nth quantity and the third auxiliary quantity, the fourth auxiliary quantity,..., the Nth auxiliary quantity.

[0084] Optionally, the step of aspirating sperm from the droplet to be aspirated according to the aspiration strategy further includes:

[0085] Calculating the third quantity + the third auxiliary quantity, and recording the result as the third simulated quantity; calculating the fourth quantity + the fourth auxiliary quantity, and recording the result as the fourth simulated quantity... calculating the Nth quantity + the Nth auxiliary quantity, and recording the result as the Nth simulated quantity;

[0086] Comparing the magnitudes of the first simulated quantity, the second simulated quantity... the Nth simulated quantity;

[0087] If the first simulated quantity is the largest, then when aspirating the droplet to be aspirated for the first time, select the first test point as the aspiration point, and when aspirating the droplet to be aspirated for the second time, select the first auxiliary point as the aspiration point; if the second simulated quantity is the largest, then when aspirating the droplet to be aspirated for the first time, select the second test point as the aspiration point, and when aspirating the droplet to be aspirated for the second time, select the second auxiliary point as the aspiration point... if the Nth simulated quantity is the largest, then when aspirating the droplet to be aspirated for the first time, select the Nth test point as the aspiration point, and when aspirating the droplet to be aspirated for the second time, select the Nth auxiliary point as the aspiration point.

[0088] The present invention has the following beneficial effects:

[0089] 1. For the sperm collection method for a reproductive medicine laboratory, by obtaining the three-dimensional image of the droplet to be aspirated, it is possible to more accurately understand the distribution of sperm within the droplet. This precise visualization technology can help operators identify areas where sperm are concentrated, avoiding sperm loss caused by blind aspiration. This technology can effectively improve the efficiency and accuracy of sperm collection.

[0090] 2. For the sperm collection method for a reproductive medicine laboratory, by performing surface segmentation on the three-dimensional image of the droplet to be aspirated in the first plane set, the second plane set, and the third plane set, and obtaining the intersection points formed by the planes in the first plane set, the second plane set, and the third plane set located inside the three-dimensional image of the droplet to be aspirated, which are respectively recorded as the first test point, the second test point... the Nth test point. The uniform distribution of points is not only a mark for the image, but also provides precise data support for subsequent sperm aspiration; through the uniformly distributed points, the system can optimize the selection of aspiration points based on the distribution density of sperm; during the collection process, operators can select the optimal area for aspiration according to the distribution of the point positions and sperm quantities, thus avoiding blind or excessive aspiration in local areas, ensuring the balance and efficiency of sperm collection.

[0091] 3. For the sperm collection method used in a reproductive medicine laboratory, the single aspiration volume is set to 1 milliliter, and the maximum aspiration area is calculated based on the single aspiration volume and the standard thickness. Then, a circle with an area equal to the maximum aspiration area is drawn with each test point as the center, and the number of sperm within the circle is obtained. The number of sperm within the circle represents the amount of sperm aspirated at this test point during aspiration. In this way, quantitative analysis of the sperm collection strategy can be carried out, and then the aspiration strategy can be optimized based on experimental data, improving the scientificity and reliability of experimental results.

[0092] 4. For the sperm collection method used in a reproductive medicine laboratory, before the start of aspiration, the liquids at the first test point, the second test point... the Nth test point are marked, and the real-time flow conditions of the liquids at the first test point, the second test point... the Nth test point during the aspiration process are simulated. After aspiration is completed, the computer models the shape of the remaining liquid droplets and the sperm distributed therein, and then obtains the distribution of sperm in the remaining liquid after aspiration at each test point, providing data support for the second aspiration. By simulating the process of liquid flow through the computer, the flow path of the liquid and the distribution changes of sperm therein during the aspiration process can be predicted. Moreover, by adjusting the aspiration strategy in real time, the accuracy and efficiency of the sperm collection process are improved.

[0093] 5. For the sperm collection method used in a reproductive medicine laboratory, the three-dimensional image of the first droplet is segmented by the planes in the first auxiliary plane set, the second auxiliary plane set, and the third auxiliary plane set, and the intersection points formed by the planes in the first auxiliary plane set, the second auxiliary plane set, and the third auxiliary plane set located inside the three-dimensional image of the first auxiliary droplet are obtained. Then, a circle with an area equal to the maximum aspiration area is drawn with each intersection point as the center, and the point corresponding to the circle with the largest number of sperm within the circle is taken as the second aspiration point. Uniformly distributing points is not only a mark for the image, but also provides accurate data support for subsequent sperm aspiration. Through the uniformly distributed points, the system can optimize the selection of aspiration points based on the distribution density of sperm. During the collection process, the operator can select the optimal area for aspiration according to the distribution of the point positions and the number of sperm, thus avoiding blind or excessive aspiration in local areas and ensuring the balance and efficiency of sperm collection.

[0094] 6. For the sperm collection method used in a reproductive medicine laboratory, calculate the sum of the first quantity and the first auxiliary quantity, the sum of the second quantity and the second auxiliary quantity... the sum of the A-th quantity and the A-th auxiliary quantity in sequence, and compare the magnitudes of the first simulated quantity, the second simulated quantity... the N-th simulated quantity. Select the two aspiration points corresponding to the simulated quantity with the largest value as the two aspiration points for two times, which can ensure that the sperm quantity contained in the liquid aspirated twice is the largest. By calculating multiple simulated quantities and auxiliary quantities and comparing to select the optimal aspiration points, the sperm collection effect can be maximally improved. This method ensures the efficient collection of sperm in different regions through multiple aspiration strategies, avoiding the waste of resources caused by uneven sperm distribution. BRIEF DESCRIPTION OF THE DRAWINGS

[0095] Figure 1 It is a schematic flowchart of the present invention.

[0096] Figure 2 It is a schematic diagram of sperm distribution of the present invention. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0097] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

[0098] Embodiment, referring to Figure 1 and Figure 2 , a sperm collection method used in a reproductive medicine laboratory, includes:

[0099] Prepare experimental mice for sperm collection;

[0100] Clean the abdomen of the experimental mice, specifically:

[0101] Use a cotton ball soaked with 75% alcohol to evenly wipe the abdomen of the experimental mice;

[0102] By cleaning the abdomen of the experimental mice, potential pollution sources can be effectively removed to ensure the accuracy of experimental results. Wiping the abdomen with 75% alcohol can not only kill surface bacteria but also reduce the interference of any microorganisms and prevent sperm contamination. And using sterilized scissors for laparotomy can further ensure the sterility of the surgical process.

[0103] After cleaning, use sterilized scissors to cut open the abdomen of the experimental mice;

[0104] Pull the peritesticular fat and separate the testis and epididymis outside the experimental mice;

[0105] Prepare two sterilized petri dishes, denoted as the first petri dish and the second petri dish respectively;

[0106] Place the testis and epididymis into the first petri dish;

[0107] Separate the epididymis from the testis in the first petri dish, and place the separated epididymis into the second petri dish;

[0108] In the second petri dish, rinse the epididymis according to the rinsing strategy, specifically:

[0109] Use a syringe to aspirate 5 ml of 0.9% normal saline that has been in a 37°C water bath;

[0110] Insert the needle of the syringe into an epididymis, and slowly push out the normal saline in the syringe so that the pushed-out normal saline forms a liquid droplet. Place the rinsed epididymis into a sterilized centrifuge tube;

[0111] Perform the same rinsing operation on the other epididymis of the experimental mouse so that the normal saline pushed out in the two rinses forms a liquid droplet;

[0112] Place the two rinsed epididymides into the same sterilized centrifuge tube;

[0113] The sterilized centrifuge tube containing the two rinsed epididymides is denoted as the target centrifuge tube;

[0114] Denote the liquid droplet formed by the normal saline pushed out in the two rinses as the liquid droplet to be aspirated;

[0115] Obtain the distribution of sperm in the liquid droplet to be aspirated;

[0116] Aspirate the sperm in the liquid droplet to be aspirated according to the aspiration strategy;

[0117] Transfer the aspirated sperm to the target centrifuge tube;

[0118] Centrifuge the target centrifuge tube to collect the sperm.

[0119] The obtaining of the distribution of sperm in the liquid droplet to be aspirated specifically includes:

[0120] Slowly move the second petri dish to the sample stage of the optical coherence tomography device, and keep the relative position between the second petri dish and the liquid droplet to be aspirated in it unchanged during the movement;

[0121] Use a low-coherence light beam to scan the liquid droplet to be aspirated layer by layer, and obtain the reflection signals at different positions of the liquid droplet to be aspirated;

[0122] According to the reflection signals at different positions within the droplet to be aspirated, the reflection signals of each layer are converted into image signals, forming a set of tomographic images ranging from light to dark;

[0123] Each layer of the image respectively corresponds to the cross-section at different positions within the droplet;

[0124] Perform image processing on each layer of the obtained image, enhance the regions with low contrast, and improve the contrast between the sperm and the surrounding background;

[0125] Synthesize each layer of the image after image processing into a three-dimensional space, denoted as the three-dimensional image of the droplet to be aspirated, and display the distribution of sperm in the three-dimensional image of the droplet to be aspirated;

[0126] This sperm collection method for reproductive medicine laboratories can more precisely understand the distribution of sperm within the droplet by obtaining the three-dimensional image of the droplet to be aspirated. This precise visualization technology can help the operator identify the regions where sperm are concentrated, avoiding sperm loss caused by blind aspiration. This technology can effectively improve the efficiency and precision of sperm collection.

[0127] The obtaining of the distribution of sperm in the droplet to be aspirated according to the sperm distribution acquisition strategy specifically includes:

[0128] Obtain the three-dimensional image of the droplet to be aspirated, and display the distribution of sperm in the three-dimensional image of the droplet to be aspirated;

[0129] Uniformly distribute points in the three-dimensional image of the droplet to be aspirated, specifically:

[0130] Use several planes parallel to the horizontal plane to divide the three-dimensional image of the droplet to be aspirated, and the distance between each plane is 0.1 millimeter. The several planes parallel to the horizontal plane form the first plane set;

[0131] Use several planes perpendicular to the planes in the first plane set to divide the three-dimensional image of the droplet to be aspirated, and the distance between each plane is 0.1 millimeter. The several planes perpendicular to the planes in the first plane set form the second plane set;

[0132] Use several planes perpendicular to the planes in both the first plane set and the second plane set to divide the three-dimensional image of the droplet to be aspirated, and the distance between each plane is 0.1 millimeter. The several planes perpendicular to the planes in both the first plane set and the second plane set form the third plane set;

[0133] The planes in the first plane set, the planes in the second plane set, and the planes in the third plane set intersect to form several intersection points;

[0134] Obtain the intersection points located inside the three-dimensional image of the droplet to be aspirated, denoted as the first test point, the second test point... the Nth test point respectively, where N represents the number of intersection points located inside the three-dimensional image of the droplet to be aspirated;

[0135] This sperm collection method for reproductive medicine laboratories divides the three-dimensional image of the droplet to be aspirated by the planes in the first plane set, the second plane set, and the third plane set, and obtains the intersection points formed by the intersection of the planes in the first plane set, the second plane set, and the third plane set inside the three-dimensional image of the droplet to be aspirated, denoted as the first test point, the second test point... the Nth test point respectively. Uniformly distributing points is not only a marking of the image, but also provides precise data support for subsequent sperm aspiration; through the uniformly distributed points, the system can optimize the selection of aspiration points based on the distribution density of sperm; during the collection process, the operator can select the optimal area for aspiration according to the distribution of the point positions and the number of sperm, thus avoiding blind or excessive aspiration in local areas and ensuring the balance and efficiency of sperm collection.

[0136] The step of aspirating sperm from the droplet to be aspirated according to the aspiration strategy further includes:

[0137] Set the single aspiration volume;

[0138] The single aspiration volume refers to the volume aspirated at one time when aspirating sperm from the droplet to be aspirated, and the single aspiration volume is equal to 1 milliliter;

[0139] Obtain the thickness of the droplet to be aspirated, denoted as the standard thickness;

[0140] Calculate single aspiration volume ÷ standard thickness, and denote the result as the maximum aspiration area;

[0141] In the three-dimensional image of the droplet to be aspirated, draw a circle with the first test point as the center and an area equal to the maximum aspiration area, denoted as the first circular surface, and the first circular surface is parallel to the horizontal plane;

[0142] Obtain the number of sperm inside the first circular surface, denoted as the first quantity;

[0143] This sperm collection method for reproductive medicine laboratories sets the single aspiration volume to 1 milliliter, calculates the maximum aspiration area based on the single aspiration volume and the standard thickness, then draws a circle with an area equal to the maximum aspiration area with each test point as the center, obtains the number of sperm inside the circle, and uses the number of sperm inside the circle to represent the amount of sperm aspirated at this test point; in this way, quantitative analysis can be carried out on the sperm collection strategy, and then the aspiration strategy can be optimized according to the experimental data, improving the scientificity and reliability of the experimental results.

[0144] The step of aspirating sperm from the droplet to be aspirated according to the aspiration strategy further includes:

[0145] When the aspiration point is the first test point and the aspiration volume is the single aspiration volume, the flow process of the liquid inside the droplet to be aspirated is simulated by computer, specifically including:

[0146] Before the aspiration starts, the liquid at the second test point, the third test point... the Nth test point is marked, and the positions of the liquid at the second test point, the third test point... the Nth test point at this time are recorded;

[0147] Obtain the moment when the aspiration starts, denoted as the start moment;

[0148] Starting from the start moment, every 0.1 ml is aspirated, and the real-time positions of the liquid at the second test point, the third test point... the Nth test point are obtained until the aspiration ends;

[0149] Based on the real-time positions of the liquid at the second test point, the third test point... the Nth test point during the aspiration process, the shape of the remaining droplet after aspiration and the sperm distributed therein are modeled, denoted as the three-dimensional image of the first auxiliary droplet;

[0150] In this sperm collection method for a reproductive medicine laboratory, before the aspiration starts, the liquid at the first test point, the second test point... the Nth test point is marked, and the real-time flow conditions of the liquid at the first test point, the second test point... the Nth test point during the aspiration process are simulated. The computer models the shape of the remaining droplet after aspiration and the sperm distributed therein, and then obtains the distribution of sperm in the remaining liquid after aspiration at each test point, providing data support for the second aspiration. By simulating the process of liquid flow by computer, the flow path of the liquid during aspiration and the distribution change of sperm therein can be predicted. Also, by adjusting the aspiration strategy in real time, this technology improves the accuracy and efficiency of the sperm collection process;

[0151] Uniformly distribute points in the three-dimensional image of the first auxiliary droplet, specifically:

[0152] Make a number of planes parallel to the horizontal plane to divide the three-dimensional image of the first auxiliary droplet, and the distance between each plane is 0.1 mm. The number of planes parallel to the horizontal plane constitutes the first auxiliary plane set;

[0153] Make a number of planes perpendicular to the planes in the first auxiliary plane set to divide the three-dimensional image of the first auxiliary droplet, and the distance between each plane is 0.1 mm. The number of planes perpendicular to the planes in the first auxiliary plane set constitutes the second auxiliary plane set;

[0154] Make several planes that are simultaneously perpendicular to the planes in the first auxiliary plane set and the second auxiliary plane set to divide the three-dimensional image of the first auxiliary droplet. The distance between each plane is 0.1 mm. The several planes that are simultaneously perpendicular to the planes in the first auxiliary plane set and the second auxiliary plane set form a third auxiliary plane set;

[0155] The planes in the first auxiliary plane set, the planes in the second auxiliary plane set, and the planes in the third auxiliary plane set intersect to form several auxiliary intersection points;

[0156] In the three-dimensional image of the first auxiliary droplet, draw a circle with an area equal to the maximum aspiration area with each auxiliary intersection point as the center. Each circular surface is parallel to the horizontal plane;

[0157] And respectively obtain the number of sperm in each circular surface;

[0158] Obtain the circular surface with the largest number of sperm, denoted as the first auxiliary circular surface;

[0159] Obtain the number of sperm in the first auxiliary circular surface, denoted as the first auxiliary quantity;

[0160] The auxiliary intersection point corresponding to the center of the first auxiliary circular surface is denoted as the first auxiliary point;

[0161] Calculate the first quantity + the first auxiliary quantity, and record the result as the first simulated quantity;

[0162] In this sperm collection method for a reproductive medicine laboratory, by making the planes in the first auxiliary plane set, the second auxiliary plane set, and the third auxiliary plane set to divide the three-dimensional image of the first droplet, obtaining the intersection points formed by the intersection of the planes in the first auxiliary plane set, the planes in the second auxiliary plane set, and the planes in the third auxiliary plane set located inside the three-dimensional image of the first auxiliary droplet, and drawing a circle with an area equal to the maximum aspiration area with each intersection point as the center, obtaining the point corresponding to the circle with the largest number of sperm inside the circle as the second aspiration point. The uniform distribution of points is not only a mark for the image, but also provides accurate data support for subsequent sperm aspiration; through the uniformly distributed points, the system can optimize the selection of aspiration points based on the distribution density of sperm; during the collection process, the operator can select the optimal area for aspiration according to the distribution of the point positions and the number of sperm, thus avoiding blind or excessive aspiration in local areas, and ensuring the balance and efficiency of sperm collection.

[0163] The sperm aspiration from the droplet to be aspirated according to the aspiration strategy further includes:

[0164] In the three-dimensional image of the droplet to be aspirated, draw a circle with an area equal to the maximum aspiration area with the second test point as the center, denoted as the second circular surface, and the second circular surface is parallel to the horizontal plane;

[0165] Obtain the number of sperm in the second circular surface, denoted as the second quantity;

[0166] When the aspiration point is the second test point and the aspiration volume is the single - aspiration volume, simulate the liquid flow process inside the liquid droplet to be aspirated through computer, specifically including:

[0167] Before the start of aspiration, mark the liquid at the first test point, the third test point... the Nth test point, and record the positions of the liquid at the first test point, the third test point... the Nth test point at this time;

[0168] Obtain the moment when aspiration starts, denoted as the start moment;

[0169] Starting from the start moment, every 0.1 milliliters of aspiration, obtain the real - time positions of the liquid at the first test point, the third test point... the Nth test point until the aspiration ends;

[0170] Model the shape of the remaining liquid droplet after aspiration and the sperm distributed therein according to the real - time positions of the liquid at the first test point, the third test point... the Nth test point during the aspiration process, denoted as the three - dimensional image of the second auxiliary liquid droplet;

[0171] Uniformly distribute points in the three - dimensional image of the second auxiliary liquid droplet, specifically:

[0172] Make a number of planes parallel to the horizontal plane to divide the three - dimensional image of the second auxiliary liquid droplet, and the distance between each plane is 0.1 millimeter. The number of planes parallel to the horizontal plane constitutes the fourth auxiliary plane set;

[0173] Make a number of planes perpendicular to the planes in the fourth auxiliary plane set to divide the three - dimensional image of the second auxiliary liquid droplet, and the distance between each plane is 0.1 millimeter. The number of planes perpendicular to the planes in the fourth auxiliary plane set constitutes the fifth auxiliary plane set;

[0174] Make a number of planes perpendicular to the planes in both the fourth auxiliary plane set and the fifth auxiliary plane set to divide the three - dimensional image of the second auxiliary liquid droplet, and the distance between each plane is 0.1 millimeter. The number of planes perpendicular to the planes in both the fourth auxiliary plane set and the fifth auxiliary plane set constitutes the sixth auxiliary plane set;

[0175] The planes in the fourth auxiliary plane set, the planes in the fifth auxiliary plane set, and the planes in the sixth auxiliary plane set intersect to form a number of auxiliary intersection points;

[0176] In the three - dimensional image of the second auxiliary liquid droplet, draw circles with an area equal to the maximum aspiration area with each auxiliary intersection point as the center, and each circular surface is parallel to the horizontal plane;

[0177] And respectively obtain the number of sperm in each circular surface;

[0178] Obtain the circular surface with the largest number of sperm, denoted as the second auxiliary circular surface;

[0179] Obtain the number of sperm in the second auxiliary circular surface, denoted as the second auxiliary quantity;

[0180] The auxiliary intersection point corresponding to the center of the first auxiliary circular surface is denoted as the first auxiliary point;

[0181] Calculate the second quantity + the second auxiliary quantity, and denote the result as the second simulated quantity;

[0182] Through the above method, respectively obtain the third quantity, the fourth quantity... the Nth quantity and the third auxiliary quantity, the fourth auxiliary quantity... the Nth auxiliary quantity.

[0183] The step of sucking sperm in the liquid droplet to be sucked according to the sucking strategy further includes:

[0184] Calculate the third quantity + the third auxiliary quantity, and denote the result as the third simulated quantity, calculate the fourth quantity + the fourth auxiliary quantity, and denote the result as the fourth simulated quantity... calculate the Nth quantity + the Nth auxiliary quantity, and denote the result as the Nth simulated quantity;

[0185] Compare the magnitudes of the first simulated quantity, the second simulated quantity... the Nth simulated quantity;

[0186] If the first simulated quantity is the largest, then when sucking the liquid droplet to be sucked for the first time, select the first test point as the sucking point, and when sucking the liquid droplet to be sucked for the second time, select the first auxiliary point as the sucking point; if the second simulated quantity is the largest, then when sucking the liquid droplet to be sucked for the first time, select the second test point as the sucking point, and when sucking the liquid droplet to be sucked for the second time, select the second auxiliary point as the sucking point... if the Nth simulated quantity is the largest, then when sucking the liquid droplet to be sucked for the first time, select the Nth test point as the sucking point, and when sucking the liquid droplet to be sucked for the second time, select the Nth auxiliary point as the sucking point;

[0187] For the sperm collection method used in a reproductive medicine laboratory, calculate the first quantity + the first auxiliary quantity, the second quantity + the second auxiliary quantity... the A quantity + the A auxiliary quantity in sequence, and compare the magnitudes of the first simulated quantity, the second simulated quantity... the Nth simulated quantity, and select the two sucking points corresponding to the largest simulated quantity as the two sucking points for two times, which can ensure that the number of sperm contained in the liquid sucked twice is the largest. By calculating multiple simulated quantities and auxiliary quantities and comparing to select the optimal sucking point, the sperm collection effect can be maximally improved. This method ensures the efficient collection of sperm in different regions through multiple sucking strategies, avoiding resource waste caused by uneven sperm distribution.

[0188] It should be noted that in this text, relational terms such as first and second are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any actual relationship or order between these entities or operations. Moreover, the terms "comprising", "including" or any other variant thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device comprising a series of elements not only includes those elements, but also includes other elements not expressly listed, or also includes elements inherent to such process, method, article or device.

[0189] The above are only the preferred embodiments of the present invention. It should be pointed out that for those of ordinary skill in the art, without departing from the technical principle of the present invention, several improvements and refinements can be made, and these improvements and refinements should also be regarded as the protection scope of the present invention.

Claims

1. A sperm collection method for reproductive medicine laboratories, characterized in that, Including: Experimental mice prepared for sperm collection; Clean the abdomen of the experimental mice, specifically: Use a cotton ball soaked with 75% alcohol to evenly wipe the abdomen of the experimental mice; After cleaning, use sterilized scissors to cut open the abdominal cavity of the experimental mice; Pull the peritesticular fat and separate the testis and epididymis outside the experimental mice; Prepare two sterilized culture dishes, denoted as the first culture dish and the second culture dish respectively; Place the testis and epididymis in the first culture dish; Separate the epididymis from the testis in the first culture dish and place the separated epididymis in the second culture dish; In the second culture dish, rinse the epididymis according to the rinsing strategy, specifically: Use a syringe to aspirate 5 ml of 0.9% physiological saline that has been in a 37°C water bath; Insert the syringe needle into an epididymis and slowly push out the physiological saline in the syringe so that the pushed-out physiological saline forms a droplet, and place the rinsed epididymis in a sterilized centrifuge tube; Perform the same rinsing operation on the other epididymis of the experimental mice so that the physiological saline pushed out in the two rinses forms a droplet; Place the two rinsed epididymides in the same sterilized centrifuge tube; The sterilized centrifuge tube containing the two rinsed epididymides is denoted as the target centrifuge tube; Denote the droplet formed by the physiological saline pushed out in the two rinses as the droplet to be aspirated; Obtain the sperm distribution in the droplet to be aspirated; Aspirate the sperm in the droplet to be aspirated according to the aspiration strategy; Transfer the aspirated sperm to the target centrifuge tube; Centrifuge the target centrifuge tube to collect sperm.

2. The sperm collection method for a reproductive medicine laboratory according to claim 1, wherein: The obtaining of the sperm distribution in the droplet to be aspirated according to the sperm distribution obtaining strategy specifically includes: Obtain a three-dimensional image of the droplet to be aspirated and display the sperm distribution in the three-dimensional image of the droplet to be aspirated; Evenly distribute points in the three-dimensional image of the droplet to be aspirated, specifically: Make several planes parallel to the horizontal plane to divide the three-dimensional image of the droplet to be aspirated, and the distance between each plane is 0.1 mm. The several planes parallel to the horizontal plane form the first plane set; Make several planes perpendicular to the planes in the first plane set to divide the three-dimensional image of the droplet to be aspirated, and the distance between each plane is 0.1 mm. The several planes perpendicular to the planes in the first plane set form the second plane set; Make several planes perpendicular to the planes in the first plane set and the second plane set to divide the three-dimensional image of the droplet to be aspirated, and the distance between each plane is 0.1 mm. The several planes perpendicular to the planes in the first plane set and the second plane set form the third plane set; The planes in the first plane set, the planes in the second plane set, and the planes in the third plane set intersect to form several intersection points; Obtain the intersection points located inside the three-dimensional image of the droplet to be aspirated, denoted as the first test point, the second test point... the Nth test point respectively, where N represents the number of intersection points located inside the three-dimensional image of the droplet to be aspirated.

3. The sperm collection method for a reproductive medicine laboratory according to claim 2, characterized in that: The aspirating of the sperm in the droplet to be aspirated according to the aspiration strategy further includes: Set the single aspiration volume; The single aspiration volume refers to the volume aspirated at one time when aspirating sperm in the liquid droplet to be aspirated, and the single aspiration volume is equal to 1 milliliter; Obtain the thickness of the liquid droplet to be aspirated, denoted as the standard thickness; Calculate the single aspiration volume ÷ the standard thickness, and denote the result as the maximum aspiration area; In the three-dimensional image of the liquid droplet to be aspirated, draw a circle with the first test point as the center and an area equal to the maximum aspiration area, denoted as the first circular surface, and the first circular surface is parallel to the horizontal plane; Obtain the number of sperm in the first circular surface, denoted as the first quantity.

4. A sperm collection method for a reproductive medicine laboratory according to claim 3, characterized in that: The step of aspirating sperm in the liquid droplet to be aspirated according to the aspiration strategy further includes: When the aspiration point is the first test point and the aspiration volume is the single aspiration volume, simulate the flow process of the liquid inside the liquid droplet to be aspirated through computer, specifically including: Before the aspiration starts, mark the liquid at the second test point, the third test point... the Nth test point, and record the positions of the liquid at the second test point, the third test point... the Nth test point at this time; Obtain the moment when the aspiration starts, denoted as the start moment; Starting from the start moment, every time 0.1 milliliter is aspirated, obtain the real-time positions of the liquid at the second test point, the third test point... the Nth test point until the aspiration ends; Model the shape of the remaining liquid droplet after aspiration and the sperm distributed therein according to the real-time positions of the liquid at the second test point, the third test point... the Nth test point during the aspiration process, denoted as the three-dimensional image of the first auxiliary liquid droplet; Uniformly distribute points in the three-dimensional image of the first auxiliary liquid droplet, specifically: Make several planes parallel to the horizontal plane to divide the three-dimensional image of the first auxiliary liquid droplet, and the distance between each plane is 0.1 millimeter. The several planes parallel to the horizontal plane form the first set of auxiliary planes; Make several planes perpendicular to the planes in the first set of auxiliary planes to divide the three-dimensional image of the first auxiliary liquid droplet, and the distance between each plane is 0.1 millimeter. The several planes perpendicular to the planes in the first set of auxiliary planes form the second set of auxiliary planes; Make several planes perpendicular to the planes in both the first set of auxiliary planes and the second set of auxiliary planes to divide the three-dimensional image of the first auxiliary liquid droplet, and the distance between each plane is 0.1 millimeter. The several planes perpendicular to the planes in both the first set of auxiliary planes and the second set of auxiliary planes form the third set of auxiliary planes; The planes in the first set of auxiliary planes, the planes in the second set of auxiliary planes, and the planes in the third set of auxiliary planes intersect to form several auxiliary intersection points; In the three-dimensional image of the first auxiliary liquid droplet, draw a circle with each auxiliary intersection point as the center and an area equal to the maximum aspiration area, and each circular surface is parallel to the horizontal plane; And respectively obtain the number of sperm in each circular surface; Obtain the circular surface with the largest number of sperm, denoted as the first auxiliary circular surface; Obtain the number of sperm in the first auxiliary circular surface, denoted as the first auxiliary quantity; The auxiliary intersection point corresponding to the center of the first auxiliary circular surface is denoted as the first auxiliary point; Calculate the first quantity + the first auxiliary quantity, and denote the result as the first simulated quantity.

5. A sperm collection method for a reproductive medicine laboratory according to claim 4, characterized in that: The step of aspirating sperm in the liquid droplet to be aspirated according to the aspiration strategy further includes: In the three-dimensional image of the droplet to be aspirated, draw a circle with the second test point as the center and an area equal to the maximum aspiration area, denoted as the second circular surface, and the second circular surface is parallel to the horizontal plane; Obtain the number of sperm within the second circular surface, denoted as the second quantity; When the aspiration point is the second test point and the aspiration volume is the single aspiration volume, simulate the flow process of the liquid inside the droplet to be aspirated through computer simulation, specifically including: Before the aspiration starts, mark the liquid at the first test point, the third test point... the Nth test point, and record the positions of the liquid at the first test point, the third test point... the Nth test point at this time; Obtain the moment when the aspiration starts, denoted as the start moment; Starting from the start moment, every 0.1 milliliters of aspiration, obtain the real-time positions of the liquid at the first test point, the third test point... the Nth test point until the aspiration ends; Based on the real-time positions of the liquid at the first test point, the third test point... the Nth test point during the aspiration process, model the shape of the remaining droplet after aspiration and the sperm distributed therein, denoted as the three-dimensional image of the second auxiliary droplet; Uniformly distribute points in the three-dimensional image of the second auxiliary droplet, specifically: Make a number of planes parallel to the horizontal plane to divide the three-dimensional image of the second auxiliary droplet, and the distance between each plane is 0.1 millimeter. The number of planes parallel to the horizontal plane constitutes the fourth auxiliary plane set; Make a number of planes perpendicular to the planes in the fourth auxiliary plane set to divide the three-dimensional image of the second auxiliary droplet, and the distance between each plane is 0.1 millimeter. The number of planes perpendicular to the planes in the fourth auxiliary plane set constitutes the fifth auxiliary plane set; Make a number of planes perpendicular to the planes in both the fourth auxiliary plane set and the fifth auxiliary plane set to divide the three-dimensional image of the second auxiliary droplet, and the distance between each plane is 0.1 millimeter. The number of planes perpendicular to the planes in both the fourth auxiliary plane set and the fifth auxiliary plane set constitutes the sixth auxiliary plane set; The planes in the fourth auxiliary plane set, the planes in the fifth auxiliary plane set, and the planes in the sixth auxiliary plane set intersect to form a number of auxiliary intersection points; In the three-dimensional image of the second auxiliary droplet, draw a circle with each auxiliary intersection point as the center and an area equal to the maximum aspiration area, and each circular surface is parallel to the horizontal plane; And respectively obtain the number of sperm in each circular surface; Obtain the circular surface with the largest number of sperm, denoted as the second auxiliary circular surface; Obtain the number of sperm in the second auxiliary circular surface, denoted as the second auxiliary quantity; The auxiliary intersection point corresponding to the center of the first auxiliary circular surface is denoted as the first auxiliary point; Calculate the second quantity + the second auxiliary quantity, and denote the result as the second simulated quantity; Through the above method, respectively obtain the third quantity, the fourth quantity... the Nth quantity and the third auxiliary quantity, the fourth auxiliary quantity... the Nth auxiliary quantity.

6. A sperm collection method for a reproductive medicine laboratory according to claim 5, characterized in that: The aspirating sperm in the droplet to be aspirated according to the aspiration strategy further includes: Calculate the third quantity + the third auxiliary quantity, and denote the result as the third simulated quantity, calculate the fourth quantity + the fourth auxiliary quantity, and denote the result as the fourth simulated quantity... calculate the Nth quantity + the Nth auxiliary quantity, and denote the result as the Nth simulated quantity; Compare the magnitudes of the first simulated quantity, the second simulated quantity, ……, the Nth simulated quantity; If the first simulated quantity is the largest, then when sucking the droplet to be sucked for the first time, select the first test point as the sucking point, and when sucking the droplet to be sucked for the second time, select the first auxiliary point as the sucking point; if the second simulated quantity is the largest, then when sucking the droplet to be sucked for the first time, select the second test point as the sucking point, and when sucking the droplet to be sucked for the second time, select the second auxiliary point as the sucking point …… if the Nth simulated quantity is the largest, then when sucking the droplet to be sucked for the first time, select the Nth test point as the sucking point, and when sucking the droplet to be sucked for the second time, select the Nth auxiliary point as the sucking point.

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