Customized micropipettor fluid modeling methods, apparatus, and media for organoid operations

Abstract

The invention discloses a customized micro-pipettor fluid modeling method, equipment and medium for organoid operation, and belongs to the technical field of micro-pipettor modeling. The invention aims to solve the problem that the motion law of internal fluid is not accurately described due to the deviation of the existing fluid model description method of the customized micro pipettor. The method comprises the following steps: acquiring a micro-pipettor source image and processing to obtain a micro-pipettor main body contour; carrying out pose correction on the main body contour; performing polynomial fitting to obtain a main body contour fitting curve under a rectangular coordinate system, and converting the main body contour fitting curve into a geometric model of the customized micro pipettor with an actual size; performing fluid static and dynamic simulation by using the geometric model, and obtaining a theoretical fluid flow velocity rule of the customized micro pipettor according to a simulation result; performing a fluid tracking experiment by using microbeads to obtain an actual fluid flow velocity rule; and verifying the theoretical fluid flow velocity rule by adopting an actual fluid flow velocity rule, and obtaining a finite element fluid simulation model based on a verification result. The method is used for fluid modeling of the micro pipettor.

Description

Technical Field

[0001] The invention relates to a customized micropipette fluid modeling method, equipment and medium for organoid operation, and belongs to the technical field of micropipette modeling. Background Art

[0002] Capillary glass needles, also known as capillary microelectrodes or micropipettes, are made mainly of silicate. Due to their light transmittance and good processability, they are widely used in micromanipulation and neuroscience research. In recent years, the development of organoid technology has further promoted the application of micropipettes, especially for different organoid operation scenarios, such as organoid injection, splicing and transfer, etc., micropipettes of different scales and shapes are processed and used. Therefore, compared with the fixed parameter micropipettes used in traditional commercialization, customized micropipettes are more flexible in size parameters, but also bring challenges to modeling in micropipette applications, especially for automated pipetting operations in automated organoid operations. More accurate modeling of micropipettes can better describe the operation process and help improve operation accuracy.

[0003] In other fields different from traditional micromanipulation, such as when operating on model animals such as cells, zebrafish, and nematodes, the micropipettes used include commercial micropipettes with fixed geometric parameters and customized micropipettes. In terms of geometric models, they usually choose to use straight line contours to approximate the curve contours of the actual model. In terms of fluid models, both of them choose general fluid models under specific conditions for approximation to study. When using micropipettes with fixed geometric parameters, their parameters lack flexibility and it is difficult to meet the increasingly complex organoid operation tasks. For example, organoid injection requires the end size of the micropipette to be several microns to more than ten microns, and organoid transfer requires the end size of the micropipette to be tens to hundreds of microns. Existing specifications of pipettes are difficult to meet the needs of all specific scenarios. When using customized micropipettes, approximate geometric models and general fluid models are often used to describe them. Although this description method greatly simplifies the modeling process, there are deviations in its description of specific operating environments. In theory, it is difficult to accurately predict the motion trajectory of the operating object, which will ultimately affect the accuracy and efficiency of automated operations of in vitro biological models represented by organoids.

[0004] With the diversification of micropipette application scenarios, micropipette with fixed specifications and sizes can no longer meet the needs of use. Although customized micropipette has largely solved this problem, there is still a lack of a method that can accurately describe the movement of fluid inside customized micropipette, which greatly affects the accuracy and efficiency of automated micromanipulation represented by organoids. Summary of the invention

[0005] In view of the problem that the fluid model description method of the existing customized micropipette has deviations, resulting in inaccurate description of the internal fluid movement law, a customized micropipette fluid modeling method, equipment and medium for organoid operation are provided.

[0006] A customized micropipette fluid modeling method for organoid manipulation of the present invention comprises:

[0007] The source image of the customized micropipette is collected and image processed to obtain the main body contour of the customized micropipette; the posture of the main body contour is calculated and the posture is corrected so that the axis of the corrected main body contour coincides with the X-axis of the rectangular coordinate system;

[0008] Perform polynomial fitting on the contour pixel points of the corrected main body contour to obtain the main body contour fitting curve in the rectangular coordinate system; convert the pixel size of the main body contour fitting curve into the actual size to obtain the geometric model of the customized micropipette;

[0009] The geometric model is used to perform static and dynamic simulations of the fluid, and the theoretical fluid flow velocity law of the customized micropipette is obtained from the static and dynamic simulation results. Microbeads are then used to perform fluid tracking experiments on the customized micropipette to obtain the actual fluid flow velocity law of the customized micropipette. The actual fluid flow velocity law is used to verify the theoretical fluid flow velocity law, and a finite element fluid simulation model is obtained based on the verification results.

[0010] According to the method for fluid modeling of a customized micropipette for organoid manipulation of the present invention, the customized micropipette is obtained by a drawing instrument or by hand drawing, and the material is transparent borate or silicate;

[0011] The source image is an axial cross-sectional image of a customized micropipette.

[0012] According to the customized micropipette fluid modeling method for organoid manipulation of the present invention, the method for obtaining the main body contour of the customized micropipette includes:

[0013] Gaussian blur is used to suppress noise in the source image, and then the region of interest is extracted; the Canny operator is used to detect the contour of the region of interest, and then the morphological closing operation is used on the contour detection result to eliminate the holes in the edge and smooth the contour edge, and the main contour is obtained after short contour filtering.

[0014] According to the customized micropipette fluid modeling method for organoid manipulation of the present invention, the method for calculating the posture of the subject contour and performing posture correction includes:

[0015] The geometric center image coordinates and the overall rotation angle of the subject contour are calculated, and the posture of the subject contour is corrected according to the geometric center image coordinates and the overall rotation angle.

[0016] According to the customized micropipette fluid modeling method for organoid manipulation of the present invention, the theoretical fluid flow rate law and the actual fluid flow rate law both include the flow rate law of each point on the axis of the customized micropipette.

[0017] According to the customized micropipette fluid modeling method for organoid manipulation of the present invention, the size and mass of the microbeads are negligible;

[0018] Methods for fluid tracking using beads for custom micropipettes include:

[0019] The template difference method is used to extract the moving micro-bead target, and the background noise of the micro-bead target is suppressed by median filtering. Then, the Kalman filter is designed based on the constant acceleration model to track the micro-bead target.

[0020] According to the customized micropipette fluid modeling method for organoid manipulation of the present invention, obtaining a finite element fluid simulation model based on the verification results includes:

[0021] The actual fluid flow velocity law is used to verify the theoretical fluid flow velocity law. If the degree of fit between the actual fluid flow velocity law and the theoretical fluid flow velocity law meets the preset threshold, the finite element fluid simulation model is determined based on the theoretical fluid flow velocity law; otherwise, the theoretical fluid flow velocity law is corrected based on the actual fluid flow velocity law, and the finite element fluid simulation model is determined based on the correction result.

[0022] According to the customized micropipette fluid modeling method for organoid manipulation of the present invention, the customized micropipette is a micropipette with a pointed end;

[0023] The outer wall profile of one side of the main body contour fitting curve is represented as f1′(x), and the inner wall profile is represented as f2′(x); the outer wall profile of the other side wall is represented as f4′(x), and the inner wall profile is represented as f3′(x); f1′(x) and f4′(x) are symmetrical profiles, and f2′(x) and f3′(x) are symmetrical profiles;

[0024] The geometric models corresponding to f1′(x), f2′(x), f3′(x) and f4′(x) are represented as f1(x), f2(x), f3(x) and f4(x) respectively:

[0025] f1(x)≈-f4(x)=a1·x 3 -b1·x 2 +c1·x+d1,

[0026] f2(x)≈-f3(x)=a2·x 3 -b2 x 2 +c2·x+d2

[0027] Where x is the X-axis coordinate of the geometric model, x∈[a,b], a is the minimum coordinate of the geometric model, and b is the maximum coordinate of the geometric model; a1, b1, c1, and d1 are the cubic coefficient, quadratic coefficient, linear coefficient, and constant term of f1(x), respectively; a2, b2, c2, and d2 are the cubic coefficient, quadratic coefficient, linear coefficient, and constant term of f2(x), respectively.

[0028] According to the customized micropipette fluid modeling method for organoid manipulation of the present invention, the theoretical fluid flow rate law of the customized micropipette is described as:

[0029] v(x)=l(x)*v input ,

[0030] Where v(x) is the fluid velocity at each point on the axis of the customized micropipette, l(x) is the proportional function, and v input Input velocity for custom micropipette tip fluid;

[0031]

[0032] The geometric model is divided into a tip segment and a tip rear segment along the axial direction, and the axial connection point of the tip segment and the tip rear segment is taken as the 0 point of the horizontal coordinate of the rectangular coordinate system; α is the tip segment curvature adjustment coefficient, β is the tip rear segment curvature adjustment coefficient, y0 is the amplitude adjustment coefficient, and θ is the asymptote coefficient of the tip rear segment function.

[0033] The present invention also provides a computer device, comprising a memory, a processor, and a computer program stored in the memory and executable on the processor, wherein the processor implements the steps of any one of the methods when executing the computer program.

[0034] The present invention also provides a medium storing computer executable instructions, wherein the computer executable instructions are used to enable a computer to execute any one of the methods.

[0035] Beneficial effects of the present invention: The present invention firstly accurately extracts the geometric contour of the micropipette through image processing, and accurately establishes a customized micropipette fluid model through theoretical simulation and microbead tracking test, wherein the contour detection method of image extraction establishes an accurate geometric model by extracting the contour curve equation; the fluid motion description method based on the combination of theoretical simulation and microbead test more completely supports the effectiveness of the fluid model, wherein the theoretical simulation can find more common and difficult-to-test fluid laws, and has the technical characteristics of being stable, reliable and highly feasible.

[0036] The method of the present invention can accurately describe the movement patterns of fluids inside a customized micropipette, and can greatly improve the accuracy and efficiency of automated micromanipulations represented by organoids. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] Figure 1 is a flow chart of a customized micropipette fluid modeling method for organoid manipulation according to the present invention;

[0038] Figure 2 is a schematic diagram of the fitting result of the corrected body contour in the embodiment; in the figure, pixel represents pixel;

[0039] Figure 3 is a schematic diagram of the modeling process in the embodiment;

[0040] Figure 4 is a schematic diagram of using a geometric model to perform finite element fluid simulation modeling in an embodiment;

[0041] Figure 5 Schematic diagram of flow rate at each point on the central axis of the micropipette detected by static simulation in the embodiment;

[0042] Figure 6 Schematic diagram of flow velocity at each point on the central axis of the micropipette detected by dynamic simulation in the embodiment;

[0043] Figure 7 This is the relationship diagram between different inputs and flow rates at each point on the central axis of the micropipette obtained from 9 sets of static simulation experiments;

[0044] Figure 8 It is the corresponding relationship diagram of the proportional function l(x) and the position;

[0045] Fig. 9 It is a schematic diagram of flow velocity verification results of using actual fluid flow velocity law to verify theoretical fluid flow velocity law in the embodiment. DETAILED DESCRIPTION

[0046] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0047] It should be noted that, in the absence of conflict, the embodiments of the present invention and the features in the embodiments may be combined with each other.

[0048] The present invention will be further described below with reference to the accompanying drawings and specific embodiments, but they are not intended to limit the present invention.

[0049] Specific implementation method 1. Combination Figure 1 As shown, the present invention provides a customized micropipette fluid modeling method for organoid manipulation, comprising:

[0050] The source image of the customized micropipette under the microscope is collected and image processing is performed to obtain the main body contour of the customized micropipette; the posture of the main body contour is calculated and the posture correction is performed so that the axis of the corrected main body contour coincides with the X-axis of the rectangular coordinate system;

[0051] Perform polynomial fitting on the contour pixel points of the corrected main body contour to obtain the main body contour fitting curve in a rectangular coordinate system; convert the pixel size of the main body contour fitting curve into the actual size, input the main body contour fitting curve of the actual size into the modeling software, and obtain the geometric model of the customized micropipette;

[0052] Import the geometric model into the simulation software, use the geometric model to perform static and dynamic simulation of the fluid, and obtain the theoretical fluid flow rate law of the customized micropipette from the static and dynamic simulation results of the fluid; then use microbeads to perform fluid tracking experiments on the customized micropipette to obtain the actual fluid flow rate law of the customized micropipette; use the actual fluid flow rate law to verify the theoretical fluid flow rate law, and obtain the finite element fluid simulation model based on the verification results. The static simulation of the fluid is the simulation of the uniform speed state of the fluid, and the dynamic simulation of the fluid is the simulation of the variable speed state of the fluid.

[0053] Furthermore, the customized micropipette is obtained by drawing an instrument or by hand, and the material is transparent borate or silicate;

[0054] The source image is an axial cross-sectional image of a customized micropipette.

[0055] In this embodiment, the method for obtaining the main body profile of the customized micropipette includes:

[0056] The source image is subjected to noise suppression by using Gaussian blur, and then the region of interest is extracted, and the region of no concern is shielded to improve the image processing efficiency; the region of interest is subjected to edge detection, and the contour is detected by using the Canny operator, and then the contour detection result is subjected to morphological closing operation to eliminate small holes in the edge and smooth the contour edge, and the effective contour is retained by contour search and short contour filtering to obtain the main contour. The main contour includes two curve segments on the inner wall and the outer wall of the micropipette.

[0057] Methods for calculating the pose of the subject's outline and performing pose correction include:

[0058] Calculate geometric information, calculate the geometric center image coordinates and overall rotation angle of the subject contour, and correct the posture of the subject contour based on the geometric center image coordinates and the overall rotation angle; during the correction process, perform geometric transformation based on the geometric information to achieve posture correction, providing a prerequisite for extracting the rectangular coordinate system of the contour information.

[0059] The theoretical fluid flow rate law and the actual fluid flow rate law both include the flow rate law of each point on the axis of the customized micropipette.

[0060] Furthermore, the size and mass of the microbeads are negligible;

[0061] Methods for fluid tracking using beads for custom micropipettes include:

[0062] The template difference method is used to extract the moving micro-bead target, and the background noise of the micro-bead target is suppressed by median filtering. Then, a Kalman filter is designed based on the constant acceleration model to track the micro-bead target and obtain the actual fluid flow rate law.

[0063] Furthermore, the finite element fluid simulation model obtained based on the verification results includes:

[0064] The actual fluid flow velocity law is used to verify the theoretical fluid flow velocity law. If the degree of fit between the actual fluid flow velocity law and the theoretical fluid flow velocity law meets the preset threshold, the finite element fluid simulation model is determined based on the theoretical fluid flow velocity law; otherwise, the theoretical fluid flow velocity law is corrected based on the actual fluid flow velocity law, and the finite element fluid simulation model is determined based on the correction result.

[0065] As an example, the customized micropipette is a micropipette with a tip at the end;

[0066] The outer wall profile of one side of the main body contour fitting curve is represented as f1′(x), and the inner wall profile is represented as f2′(x); the outer wall profile of the other side wall is represented as f4′(x), and the inner wall profile is represented as f3′(x); f1′(x) and f4′(x) are symmetrical profiles, and f2′(x) and f3′(x) are symmetrical profiles;

[0067] The geometric models corresponding to f1′(x), f2′(x), f3′(x) and f4′(x) are represented as f1(x), f2(x), f3(x) and f4(x) respectively:

[0068] f1(x)≈-f4(x)=a1·x 3 -b1·x 2 +c1·x+d1,

[0069] f2(x)≈-f3(x)=a2·x 3 -b2 x 2 +c2·x+d2

[0070] Where x is the X-axis coordinate of the geometric model, x∈[a,b], a is the minimum coordinate of the geometric model, and b is the maximum coordinate of the geometric model; a1, b1, c1, and d1 are the cubic coefficient, quadratic coefficient, linear coefficient, and constant term of f1(x), respectively; a2, b2, c2, and d2 are the cubic coefficient, quadratic coefficient, linear coefficient, and constant term of f2(x), respectively.

[0071] In this embodiment, the theoretical fluid flow rate law of the customized micropipette is described as:

[0072] v(x)=l(x)*v input ,

[0073] Where v(x) is the fluid velocity at each point on the axis of the customized micropipette, l(x) is the proportional function, and v input Input velocity for custom micropipette tip fluid;

[0074]

[0075] The geometric model is divided into a tip segment and a tip rear segment along the axial direction, and the axial connection point of the tip segment and the tip rear segment is taken as the 0 point of the horizontal coordinate of the rectangular coordinate system; α is the tip segment curvature adjustment coefficient, β is the tip rear segment curvature adjustment coefficient, y0 is the amplitude adjustment coefficient, and θ is the asymptote coefficient of the tip rear segment function.

[0076] Example:

[0077] The beneficial effects of the present invention are further illustrated below by means of specific examples, which on the one hand prove the feasibility of the method of the present invention, and on the other hand reflect the advantages of the method of the present invention through comparison.

[0078] Take a silicate micropipette with an outer diameter of 1 mm and an inner diameter of 0.5 mm as an example:

[0079] 1) Use a commercial needle puller Sutter P97 to set the parameters HEAT=500, PULL=0, VEL=130, TIME=200 to pull a micropipette, and the tip size of the grinded pipette is about 110 μm;

[0080] 2) Obtain the source image of the micropipette tip profile under a microscope (8192*6144 pixels);

[0081] 3) Use Gaussian blur source image, kernel size: 51*51;

[0082] 4) Extracting regions of interest based on image features;

[0083] 5) Use Canny edge detection to detect image edges with double threshold (10, 20);

[0084] 6) Perform morphological closing operation, kernel size: 9*9;

[0085] 7) Use contour length to filter out short contours and retain the main contours, threshold: 4000;

[0086] 8) Calculate the geometric center of the contour, and average the rotation angle of each contour to obtain the overall contour rotation angle;

[0087] 9) Adjust the contour posture according to the above posture information; obtain the fitting result of the corrected subject contour as follows Figure 2 As shown;

[0088] 10) Fit the contour data points and obtain the actual contour curve, i.e., the geometric model, according to the ratio of pixel size to actual size. The curve equation is finally obtained as:

[0089] f1(x)≈-f4(x)=5.50×10 -8 ·x 3 -5.17×10 -6 ·x 2 +0.21 x+242.11,

[0090] f2(x)≈-f3(x)=3.27×10 -8 ·x 3 -6.88×10 -6 ·x 2 +0.13 x+161.71

[0091] Where x is the X-axis coordinate of the geometric model, x∈[-958,1150].

[0092] 11) Use equation-driven modeling methods in modeling software to obtain geometric models, such as Figure 3 As shown, including (a) micropipette source image, (b) closing operation processing result, (c) main contour extraction result and (d) modeling result;

[0093] 12) Use geometric models to perform fluid finite element simulation, including dynamic simulation and static simulation. Figure 4 shown.

[0094] 13) In the static simulation, a constant flow rate input is applied to the end of the pipette, and then the flow rate at each point on the central axis of the micropipette is detected, such as Figure 5 As shown in the figure; in the dynamic simulation, a sinusoidal flow rate input is applied to the end of the pipette, and then the flow rate changes at each point in the center of the micropipette are detected, as shown in the figure. Figure 6 As shown; 9 groups of static simulation experiments were repeated, each group of experiments applied different inputs, and the flow rate at different points was detected to determine that the relationship between input and output was a linear relationship, such as Figure 7As shown; piecewise functions are used to describe the fluid law, and the fluid law is described as:

[0095] v(x)=l(x)*v input ;

[0096] The proportional function l(x) reflects the linear relationship between the flow velocity at each point on the streamline and the input, such as Figure 8 shown.

[0097]

[0098] For the micropipette with a 130 μm nozzle, the values ​​of α, β, y0, and θ are 6288, -956.5, 17.06, and -1.806, respectively.

[0099] 14) Use 5 μm polystyrene beads as tracking objects to verify and correct the simulation fluid model, such as Fig. 9 shown.

[0100] 15) In this example, the normalized standard deviation of the simulation and test is 0.0174, which proves the validity of the simulation results. No correction is required in this case.

[0101] Exemplary computer devices of the present invention:

[0102] The device includes a memory, a processor, and a computer program stored in the memory and executable on the processor. When the processor executes the computer program, the steps of any method described in the specific implementation manner are implemented.

[0103] Exemplary media of the present invention:

[0104] The medium stores computer-executable instructions, and the computer-executable instructions are used to enable a computer to execute any method described in any specific implementation manner.

[0105] In the present invention, automated micromanipulation refers to a technology that performs microscopic scale operations through computer control and precision mechanical equipment, with particular emphasis on the use of image servo systems to monitor and adjust operations in real time. This method can be used for experiments such as cell manipulation and microinjection, improves the accuracy and efficiency of the experiment, reduces human errors, and is widely used in biomedical research and nanotechnology. Organoids are three-dimensional cell structures cultured in the laboratory that can simulate the function and tissue structure of natural organs. They are generated from stem cells or primary cells and are widely used in disease research, drug screening, and regenerative medicine.

[0106] In summary, the method of the present invention can describe the fluid movement law in the tube of a customized micropipette when performing an operation task. The geometric modeling process based on image processing can provide a general method for modeling pipettes of different shapes and specifications, and has the possibility of being integrated into automated micromanipulation instruments. In fluid modeling, theory plus experimental verification more effectively supports the effectiveness and reliability of the description of fluid laws. The principle on which the description method is based is simple, the description process is intuitive and easy to understand, and can be well integrated into automated micromanipulation.

[0107] The method of the present invention obtains the three-dimensional morphology of the micropipette entity based on image contour curve fitting. The method can more intuitively extract the morphological features of the micropipette, and makes good use of the good light transmittance of the micropipette, so the modeling process is very direct and accurate. In fluid modeling, compared with existing methods (using general fluid formulas), it is more accurate when targeting specific scenes. The simulation method based on finite element theory provides an important reference for the law of fluid motion under complex environments, especially in some environments where it is difficult to perform experimental measurements. The method of verifying the fluid model by microbead tracking once again utilizes the good light transmittance of the micropipette and the mature characteristics of the image-based target tracking technology. Therefore, the principle is simple, the description process is intuitive, and it has engineering practical value. It can be better applied to the fluid modeling description of customized micropipette, especially for the complex tasks and the cross-scale organoid micromanipulation. It is very meaningful.

[0108] Although the present invention is described herein with reference to specific embodiments, it should be understood that these embodiments are merely examples of the principles and applications of the present invention. It should therefore be understood that many modifications may be made to the exemplary embodiments and that other arrangements may be devised without departing from the spirit and scope of the present invention as defined by the appended claims. It should be understood that the various dependent claims and features described herein may be combined in a manner different from that described in the original claims. It should also be understood that the features described in conjunction with a single embodiment may be used in other described embodiments.

Claims

1. A customized micropipette fluid modeling method for organoid manipulation, characterized in that: include: Acquiring a source image of the customized micropipette and performing image processing to obtain a main outline of the customized micropipette; Calculate the posture of the subject's contour and perform posture correction so that the axis of the corrected subject's contour coincides with the X-axis of the rectangular coordinate system; Perform polynomial fitting on the contour pixel points of the corrected main body contour to obtain the main body contour fitting curve in the rectangular coordinate system; convert the pixel size of the main body contour fitting curve into the actual size to obtain the geometric model of the customized micropipette; The geometric model is used to perform static and dynamic simulations of the fluid, and the theoretical fluid flow velocity law of the customized micropipette is obtained from the static and dynamic simulation results. Microbeads are then used to perform fluid tracking experiments on the customized micropipette to obtain the actual fluid flow velocity law of the customized micropipette. The actual fluid flow velocity law is used to verify the theoretical fluid flow velocity law, and a finite element fluid simulation model is obtained based on the verification results.

2. The customized micropipette fluid modeling method for organoid manipulation according to claim 1, characterized in that: The customized micropipette is obtained by drawing an instrument or by hand, and the material is transparent borate or silicate; The source image is an axial cross-sectional image of a customized micropipette.

3. The customized micropipette fluid modeling method for organoid manipulation according to claim 2, characterized in that: Methods for obtaining a custom micropipette body profile include: Gaussian blur is used to suppress noise in the source image, and then the region of interest is extracted; the Canny operator is used to detect the contour of the region of interest, and then the morphological closing operation is used on the contour detection result to eliminate the holes in the edge and smooth the contour edge, and the main contour is obtained after short contour filtering.

4. The customized micropipette fluid modeling method for organoid manipulation according to claim 3, characterized in that: Methods for calculating the pose of the subject's outline and performing pose correction include: The geometric center image coordinates and the overall rotation angle of the subject contour are calculated, and the posture of the subject contour is corrected according to the geometric center image coordinates and the overall rotation angle.

5. The customized micropipette fluid modeling method for organoid manipulation according to claim 4, characterized in that: The theoretical fluid flow rate law and the actual fluid flow rate law both include the flow rate law of each point on the axis of the customized micropipette.

6. The customized micropipette fluid modeling method for organoid manipulation according to claim 5, characterized in that: The size and mass of the microbeads are negligible; Methods for fluid tracking using beads for custom micropipettes include: The template difference method is used to extract the moving microbead targets, and the background noise of the microbead targets is suppressed by median filtering. Then, a Kalman filter is designed based on the constant acceleration model to track the microbead target.

7. The customized micropipette fluid modeling method for organoid manipulation according to claim 6, characterized in that: The finite element fluid simulation model based on the verification results includes: The actual fluid flow velocity law is used to verify the theoretical fluid flow velocity law. If the degree of fit between the actual fluid flow velocity law and the theoretical fluid flow velocity law meets the preset threshold, the finite element fluid simulation model is determined based on the theoretical fluid flow velocity law; otherwise, the theoretical fluid flow velocity law is corrected based on the actual fluid flow velocity law, and the finite element fluid simulation model is determined based on the correction result.

8. The customized micropipette fluid modeling method for organoid manipulation according to claim 7, characterized in that: The customized micropipette is a micropipette with a pointed tip at the end; The outer wall profile of one side of the main body contour fitting curve is represented as f1′(x), and the inner wall profile is represented as f2′(x); the outer wall profile of the other side wall is represented as f4′(x), and the inner wall profile is represented as f3′(x); f1′(x) and f4′(x) are symmetrical profiles, and f2′(x) and f3′(x) are symmetrical profiles; The geometric models corresponding to f1′(x), f2′(x), f3′(x) and f4′(x) are represented as f1(x), f2(x), f3(x) and f4(x) respectively: f1(x)≈-f4(x)=a1·x 3 -b1·x 2 +c1·x+d1, f2(x)≈-f3(x)=a2·x 3 -b2·x 2 +c2·x+d2 Where x is the X-axis coordinate of the geometric model, x∈[a,b], a is the minimum coordinate of the geometric model, and b is the maximum coordinate of the geometric model; a1, b1, c1, and d1 are the cubic coefficient, quadratic coefficient, linear coefficient, and constant term of f1(x), respectively; a2, b2, c2, and d2 are the cubic coefficient, quadratic coefficient, linear coefficient, and constant term of f2(x), respectively.

9. The customized micropipette fluid modeling method for organoid manipulation according to claim 8, characterized in that: The theoretical fluid flow rate law of the customized micropipette is described as: v(x)=l(x)*v input , Where v(x) is the fluid velocity at each point on the axis of the customized micropipette, l(x) is the proportional function, and v input Input velocity for custom micropipette tip fluid; The geometric model is divided into a tip segment and a tip rear segment along the axial direction, and the axial connection point of the tip segment and the tip rear segment is taken as the 0 point of the horizontal coordinate of the rectangular coordinate system; α is the tip segment curvature adjustment coefficient, β is the tip rear segment curvature adjustment coefficient, y0 is the amplitude adjustment coefficient, and θ is the asymptote coefficient of the tip rear segment function.

10. A computer device comprising a memory, a processor, and a computer program stored in the memory and executable on the processor, characterized in that: When the processor executes the computer program, the steps of the method according to any one of claims 1 to 9 are implemented.

11. A medium, characterized in that The medium stores computer-executable instructions, and the computer-executable instructions are used to enable a computer to execute the method according to any one of claims 1 to 9.

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