A coating method and coating system

Abstract

The application discloses a coating method and a coating system, and relates to the technical field of microbiology, and in particular to a coating method and a coating system for a solid culture medium surface. The coating method comprises the following steps: providing a container which carries a solid culture medium surface; applying horizontal oscillation to the container, so that the solid culture medium surface is in a horizontal oscillation state; and spraying a fluid to be coated onto the solid culture medium surface while keeping the solid culture medium surface in the horizontal oscillation state, the fluid being a bacterial solution containing microorganisms. When the fluid contacts the solid culture medium surface in the horizontal oscillation state, the fluid is dispersed under the action of the horizontal oscillation and adheres to different regions of the solid culture medium surface due to adhesion, so as to form isolated single clone colonies. The application can simultaneously meet the requirements of high-throughput automation adaptability and single clone colony distribution uniformity, and further improves the single clone colony acquisition rate.

Description

Technical Field

[0001] This invention relates to the field of surface coating technology, and more specifically to a coating method and coating system. Background Technology

[0002] In the field of microbial culture and molecular cloning experiments, uniformly spreading the transformed bacterial solution onto the surface of a solid culture medium to obtain evenly distributed, relatively isolated, and non-overlapping single-clone colonies is a key prerequisite for achieving subsequent high-throughput positive clone screening and automated selection.

[0003] Traditional colony coating methods mainly include plating with a plating stick, glass bead mixing and coating, streaking, and spotting. Plating with a plating stick offers excellent coating uniformity, but it relies on repeated sterilization and cooling of the plating stick, making it slow, prone to killing bacteria, and unsuitable for batch samples. Furthermore, in automated high-throughput experiments, there is currently no mature automated plating stick replacement solution; high throughput refers to the ability to process or analyze a large number of samples per unit time.

[0004] The glass bead mixing and coating method requires the preparation of sterile glass beads and an additional operation of tilting and recovering the beads after coating. For small-diameter culture plates or multi-well plates, due to the narrow diameter and large depth of each well, glass beads are easily stuck at the corners of the well walls during tilting and are difficult to remove completely. In automated processes, existing robotic arms and gripper mechanisms are unable to perform precise and stable tilting and tilting actions for multi-well plates of different sizes. Furthermore, if glass beads become stuck, the size of existing general-purpose grippers is usually larger than the edge gripping area of ​​small-diameter plates, making it difficult to stably grasp and remove the glass beads. This results in low automation adaptability and difficulty in meeting high-throughput requirements.

[0005] Streak plating is prone to uneven streaking, resulting in low uniformity of single-clone colony distribution. It is difficult to obtain a large number of isolated single-clone colonies within a limited culture area, and satellite colonies tend to form at the ends of the streaks. Spot plating is limited by the spot volume and spacing, resulting in a small coverage area, which is difficult to meet the needs of high throughput. Furthermore, streaking or spotting tools can easily puncture the surface of the culture medium due to incompatibility or improper use, leading to sample contamination.

[0006] In the prior art, there is also a drop-and-oscillate coating method. For example, patent publication number CN115926970A discloses a coating method, including: controlling a coating liquid device to drop coating liquid into coating wells in a coating culture dish; placing the coating culture dish on a support position of an oscillator; controlling the oscillator to oscillate the coating culture dish, so that the coating liquid diffuses in the coating wells for coating. The control of the coating liquid device to drop coating liquid into the coating wells in the coating culture dish includes: controlling the coating liquid device to drop coating liquid into multiple preset positions in the coating wells.

[0007] The above coating method has the following drawbacks: 1. Because the liquid is added dropwise first and then shaken, once the stationary droplet has formed a stable contact with the surface of the culture medium, the sudden start of shaking causes the speed to accelerate instantaneously from zero to the set speed. This sudden acceleration will generate a transient inertial force, which may cause the droplet to shift, thus affecting the uniformity of the distribution of monoclonal colonies.

[0008] 2. It is necessary to drip coating liquid into multiple preset positions in the coating orifice, which is time-consuming and not conducive to meeting the requirements of high-throughput automation.

[0009] 3. Before the shaking is initiated, the droplets added statically have undergone a drying period. The evaporation rate at the edge of the droplet is higher than that at the center, causing some bacteria to move to the edge and begin to deposit. Once edge deposition is formed, subsequent shaking can only break up the bacteria that have not yet deposited, but the deposited bacteria cannot be resuspended by subsequent shaking, affecting the uniformity of monoclonal colony distribution.

[0010] In summary, existing bacterial coating methods cannot simultaneously meet the requirements of high-throughput automation and uniform distribution of single-clone colonies. Summary of the Invention

[0011] To address the shortcomings of existing technologies, this application provides a coating method and coating system that can simultaneously meet the requirements of high-throughput automation adaptability and uniform distribution of monoclonal colonies, and further improve the monoclonal colony acquisition rate.

[0012] To address the above problems, the present invention provides the following technical solution: In a first aspect, embodiments of this application provide a coating method, including: Provide a container that carries the surface of a solid culture medium; Apply horizontal agitation to the container to keep the surface of the solid culture medium in a horizontal agitation state; While maintaining horizontal agitation on the surface of the solid culture medium, the fluid to be coated is sprayed onto the surface of the solid culture medium in mid-air. The fluid is a bacterial solution containing microorganisms. When the fluid comes into contact with the surface of a solid culture medium that is in a horizontal oscillation state, it is dispersed by the horizontal oscillation and adheres to different areas of the solid culture medium surface due to adhesive force, so as to form isolated monoclonal colonies. After providing a container carrying the surface of a solid culture medium, the coating method further includes: The container is transferred from the first position to the horizontal oscillation mechanism by the first robotic arm, which is used to apply horizontal oscillation to the container; The second robotic arm transfers the sample container containing the fluid from the second position to the reach of the pipette, so that the pipette can draw the fluid from the sample container. The pipette is used to draw the fluid to be coated and spray the fluid onto the surface of the solid culture medium.

[0013] In some embodiments, the coating method further includes: The fluid is sprayed onto the surface of the solid culture medium in the form of multiple droplets, and each droplet experiences greater adhesion force than centrifugal force under horizontal oscillation, thus adhering to the vicinity of the spraying location.

[0014] In some embodiments, the container is a multi-well plate, the multi-well plate including multiple wells, each well bearing a surface of solid culture medium; while the surface of the solid culture medium is kept horizontally agitated, the fluid to be coated is suspended and sprayed onto the surface of the solid culture medium, including: While maintaining horizontal agitation on the surface of the solid culture medium, fluid is simultaneously or sequentially sprayed onto the surface of the solid culture medium in multiple wells of a multi-well plate using a pipetting device.

[0015] In some embodiments, the pipetting device includes a multi-channel robotic arm that, while maintaining horizontal oscillation on the surface of a solid culture medium, suspends and sprays the fluid to be coated onto the surface of the solid culture medium, including: A multi-channel robotic arm grasps a corresponding number of pipette tips to match the spacing between the pipette tips and the well spacing of the multi-well plate, allowing multiple pipette tips to aspirate fluid from the sample container. Move multiple pipette tips above the corresponding multiwell plate; While maintaining horizontal agitation on the surface of the solid culture medium, multiple pipette tips suspend the aspirated fluid and spray it onto the surface of the solid culture medium in the corresponding well.

[0016] In some embodiments, the volume of fluid sprayed onto the surface of the solid culture medium is 30 to 80 μL.

[0017] In some implementations, the method further includes: before the multiple pipette tips aspirate fluid from the sample container. The fluid in the sample container was mixed by blowing and sucking multiple times.

[0018] In some implementations, the method further includes: After the multi-channel robotic arm completes the suspended spraying of fluid to a set of wells in the multi-well plate, the multi-channel robotic arm removes the currently gripped pipette tip and grabs a new pipette tip. The new pipette tip performs fluid suspension spraying on another set of wells in the multi-well plate; Repeat the process of removing the suction head, grabbing the new suction head, and spraying while suspended in the air until all holes have been sprayed while suspended in the air. Multiple wells were capped, and the capped solid culture medium was transferred to an incubator for incubation.

[0019] Secondly, embodiments of this application also provide a coating system, the coating system comprising: A horizontal oscillation mechanism is used to support a container with a solid culture medium surface and to apply horizontal oscillation to the container; A pipetting device is used to pick up the fluid to be coated and spray it onto the surface of a solid culture medium while suspended in the air. The control unit, which is communicatively connected to the horizontal oscillation mechanism and the pipetting device, is used to control the activation of the horizontal oscillation mechanism to keep the surface of the solid culture medium in a horizontal oscillation state; and to control the pipetting device to spray the fluid to be coated onto the surface of the solid culture medium while keeping the surface of the solid culture medium in a horizontal oscillation state.

[0020] In some embodiments, a first robotic arm and a second robotic arm are also included, both of which are communicatively connected to the control unit; The first robotic arm is used to transfer the container from the first position to the horizontal oscillation mechanism; The second robotic arm is used to transfer the sample container containing fluid from the second position to within reach of the pipette, so that the pipette can aspirate the fluid from the sample container.

[0021] Beneficial effects: 1. This application eliminates the need for contact coating tools such as coating rods and inoculation loops, thus eliminating the need for repeated high-temperature sterilization and cooling of coating rods. This reduces the mechanical complexity and control difficulty of the automated system, and avoids problems such as glass beads getting stuck in the orifice wall and the grippers being unable to grasp and pour the contents, thereby improving the adaptability of automation.

[0022] 2. This application improves the uniformity of monoclonal colony distribution by combining suspended spraying and horizontal shaking. Suspended spraying causes the bacterial solution to fall onto the culture medium surface as multiple discrete microdroplets. The discreteness of the droplet landing points avoids the problem of bacterial accumulation caused by excessive concentration of a single droplet in traditional spotting methods, thus improving the uniformity of colony distribution.

[0023] When droplets come into contact with the surface of a culture medium that is in a horizontal oscillation state, the horizontal oscillation causes the droplets to be continuously subjected to tangential shear force during the drying process. This avoids the situation caused by droplets flowing and aggregating towards the edge during the drying process, which is caused by traditional static coating. As a result, the bacteria are randomly dispersed in the area covered by the droplets and fixed in different positions by adhesive forces, thus forming multiple isolated monoclonal colonies. Attached Figure Description

[0024] Figure 1This is a schematic flowchart of the coating method provided in the embodiments of this application.

[0025] Figure 2 This is a schematic diagram of the working state of the coating system provided in the embodiments of this application. Detailed Implementation

[0026] The technical solutions of the embodiments of this application will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this application, and not all embodiments. Based on the embodiments of this application, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of this application.

[0027] Furthermore, the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. In the description of this invention, "a plurality of" means two or more, unless otherwise explicitly specified.

[0028] The coating method provided in this application will now be described in detail with reference to the accompanying drawings.

[0029] Please see Figure 1 , Figure 1 This is a schematic flowchart of the first embodiment of the coating method provided in this application. Figure 1 As shown, the coating method includes steps S100 to S800.

[0030] Step S100: Provide a container that carries the surface of a solid culture medium.

[0031] Specifically, the container can be a multi-well plate or a standard petri dish. If it is a multi-well plate, it can be a 24-well plate, a 96-well plate, etc. In this embodiment, a multi-well plate is used. The multi-well plate includes multiple wells, each of which carries a solid culture medium surface. Specifically, each well contains liquid culture medium, which solidifies to form a smooth solid culture medium surface.

[0032] Preferably, the solid culture medium of this application is a resistant solid culture medium, with one or more antibiotics of a specific concentration added to kill or inhibit all miscellaneous bacteria / original bacteria that do not contain the gene.

[0033] In this embodiment, the container is a 24-hole plate with a lid, and the interval between two adjacent holes is 18.5 to 19.0 mm.

[0034] In some embodiments, step S100 further includes steps S110 to S120, the specific steps of which are as follows: Step S110: The container is transferred from the first position to the horizontal oscillation mechanism by the first robotic arm, which is used to apply horizontal oscillation to the container.

[0035] Optionally, the first robotic arm is an external robotic arm with grippers for grasping perforated plates. The grippers grasp the container and transfer it from the first position to the horizontal oscillation mechanism.

[0036] Optionally, the first position can be a material storage rack, a culture medium preparation area, etc., and the horizontal oscillation mechanism is a mechanism that can generate periodic motion in the horizontal direction.

[0037] Specifically, the first robotic arm receives a gripping command, moves to the first position, closes its gripper to hold the edge of the container, and then moves the container horizontally above the horizontal oscillation mechanism along a preset motion trajectory. The container is then placed on the horizontal oscillation mechanism, which is equipped with buckles or clips to fix the container in place, preventing the container from being thrown out or displaced during the oscillation process.

[0038] Step S120: The sample container carrying the fluid is transferred from the second position to the reach of the pipetting device by the second robotic arm, so that the pipetting device can draw the fluid from the sample container. The pipetting device is used to draw the fluid to be coated and spray the fluid to be coated onto the surface of the solid culture medium.

[0039] Specifically, the sample container can be a sample carrier plate, which contains fluid, specifically a bacterial solution containing microorganisms.

[0040] Optionally, the second robotic arm is an external robotic arm with grippers for grasping sample containers.

[0041] Alternatively, the second location can be a material storage rack or a sample storage area.

[0042] Specifically, upon receiving a gripping command, the second robotic arm moves to a second position, its grippers close to hold the edge of the sample container, transferring the sample container into the operating range of the pipetting device. The pipetting device has a pipette tip used to aspirate fluid from the sample container, and its operating range is the spatial area that the pipette tip can reach through its axis of motion.

[0043] Optionally, the pipetting device in this embodiment is a pipetting workstation. The pipetting workstation includes a sample holder, a multi-channel robotic arm, and a wrench-like robotic arm. The sample holder is located near and below the multi-channel robotic arm. The wrench-like robotic arm can be used to place containers on a horizontal oscillation mechanism or to place sample containers on the sample holder. Here, the first and second robotic arms can be wrench-like robotic arms. The sample container and its contents can be transferred to the pipetting workstation by an external robotic arm, and then the wrench-like robotic arm of the pipetting workstation transfers the container to the horizontal oscillation mechanism, thus transferring the sample container to the reach of the pipetting device.

[0044] In the above technical solution, steps S110 and S120 are used to automatically transfer the container to the horizontal oscillation mechanism and transfer the sample container to the reach of the pipetting device, which facilitates the automation of subsequent coating and improves the automation of the entire process.

[0045] It should be noted that the execution order of steps S110 and S120 is not strictly limited and can be executed in parallel to improve overall efficiency. These two steps achieve full automation of material feeding in the coating process without manual intervention.

[0046] Step S200: Apply horizontal shaking to the container to bring the surface of the solid culture medium into a horizontal shaking state.

[0047] Optionally, the horizontal motion can be circular or linear. Preferably, this embodiment uses a track-type horizontal oscillation mechanism, in which the fluid performing the circular motion forms a uniform annular flow within the hole, resulting in uniform shear force distribution, no splashing, and the smoothest coating.

[0048] Specifically, the horizontal oscillation mechanism can be in the form of circular oscillation with a frequency of 500-1200 revolutions per minute and an oscillation duration of 15-20 seconds. The oscillation duration can be set according to the fluid volume and the required degree of dryness of the fluid.

[0049] Specifically, the horizontal oscillation mechanism is activated, causing the entire container to vibrate in a circular motion in the horizontal plane. During the oscillation, the surface of the solid culture medium moves synchronously with the container, forming a continuous horizontal oscillation state.

[0050] Step S300: While maintaining horizontal shaking on the surface of the solid culture medium, the fluid to be coated is sprayed onto the surface of the solid culture medium in mid-air. The fluid is a bacterial solution containing microorganisms.

[0051] When a fluid comes into contact with the surface of a solid culture medium that is in a horizontal oscillation state, it is dispersed by the horizontal oscillation and adheres to different areas of the solid culture medium surface due to adhesive forces, thus forming isolated monoclonal colonies.

[0052] Specifically, the container is a multi-well plate, which includes multiple wells, each of which carries a solid culture medium surface. While the solid culture medium surface is kept horizontally agitated, the fluid is simultaneously or sequentially sprayed onto the solid culture medium surface of the multiple wells of the multi-well plate using a pipetting device.

[0053] During the continuous shaking, the multi-channel robotic arm of the pipette draws fluid from the sample container and sprays it onto the surface of the solid culture medium in a suspended spray manner. During spraying, the tip of the pipette is kept at a certain distance from the surface of the culture medium, avoiding any physical contact and thus preventing contamination of the sample.

[0054] Optionally, the distance between the tip of the pipette and the surface of the culture medium is 2 to 5 mm, which shortens the falling distance of the sprayed droplets, reduces dispersion and deviation, and sets the discharge speed of the pipette tip to a low speed to avoid the droplets from being sprayed in a mist, so that the spraying area extends beyond the surface of the culture medium.

[0055] When fluid falls as droplets onto the surface of a horizontally oscillating culture medium, the droplets are subjected to tangential shear force the instant they contact the surface, and are further dispersed into smaller units. Due to the large number of micron-sized pores on the surface of the culture medium, an adsorption force, i.e. an adhesion force, is generated between the droplets and the surface, thus attaching to different areas of the solid culture medium surface to form isolated monoclonal colonies.

[0056] In some implementations, step S300 further includes step S310.

[0057] Step S310: The fluid is sprayed onto the surface of the solid culture medium in the form of multiple droplets, and the adhesive force experienced by each droplet under horizontal oscillation is slightly greater than the centrifugal force, thus adhering to the vicinity of the spraying position.

[0058] Specifically, the volume of fluid sprayed onto the surface of the solid culture medium in each well is preferably 30 to 80 μL, more preferably 50 μL. During spraying, the center of the pipette tip is aligned directly above the center of the well, and the fluid is sprayed in the form of multiple discrete droplets within a localized area of ​​the well's central region. The initial coverage area of ​​this localized area is less than the total surface area of ​​the solid culture medium, for example, not exceeding 50%. The adhesive force experienced by each droplet is slightly greater than the centrifugal force, thus fixing the droplet's position near the spray landing point and preventing it from migrating to the well wall or other areas due to horizontal oscillation. This ensures that the bacterial solution in different landing areas is isolated from each other, forming isolated monoclonal colonies. However, although the bottom of the droplet is fixed to the culture medium surface by adhesive force, under the continuous action of horizontal oscillation, the liquid inside the droplet may disperse into smaller parts, gradually spreading outwards from the center of the droplet, thereby achieving uniform coverage of the entire well and the isolated distribution of monoclonal colonies. It should be noted that the spreading process is controllable, with the shear force driven by horizontal oscillation influencing the droplet's rolling direction, rather than the uncontrollable random rolling after the droplet splits.

[0059] In some embodiments, step S310 includes steps S311 to S313, the specific steps of which are as follows: Step S311: The pipetting device includes a multi-channel robotic arm that grasps a corresponding number of pipette tips to match the spacing between the pipette tips and the well spacing of the multi-well plate. Multiple pipette tips aspirate fluid from the sample container.

[0060] Specifically, in this embodiment, the industry standard well center-to-well spacing of the 24-well plate is 19mm, and the 24-well plate is arranged in 4 rows by 6 columns. The original channel spacing of the pipette tips in the eight-channel pipetting robot arm is typically 9mm. The multi-channel robot arm is an eight-channel robot arm, which grasps four pipette tips, that is, there is an empty channel without a pipette tip between each two adjacent pipette tips. However, there is a deviation between the four pipette tips, and longitudinal coordinate compensation needs to be applied to each channel through the robot arm control software so that the center of the four pipette tips can coincide with the center of the four wells in the same column.

[0061] Optionally, the eight-channel pipetting robot arm can be an eight-channel pipetting robot arm with adjustable spacing, which can be directly adjusted according to the needs so that the center of the four pipetting tips can coincide with the center of the four wells in the same column.

[0062] In some embodiments, before multiple pipette tips draw fluid from the sample container, the fluid in the sample container is further subjected to multiple blow-and-pipette mixing operations to ensure uniform bacterial concentration.

[0063] Specifically, the blowing and sucking mixing operation is performed 3 times.

[0064] Step S312: Move multiple pipette tips above the corresponding multiwell plate.

[0065] Specifically, the four pipette tips are moved to their respective positions so that their centers coincide with the centers of the four wells in the same row.

[0066] Under high-speed circular oscillation, such as the 800 rpm condition in this embodiment, the surface of the solid culture medium undergoes high-speed circular motion with the horizontal oscillation mechanism, while the pipette tip remains suspended in a fixed spatial position and does not rotate with the platform. The droplet falls vertically from the tip, and during its flight time, the center of the well below has shifted tangentially along the circumference with the platform. To compensate for this displacement and ensure that the droplet accurately falls into the center of the well, step S312 in this embodiment includes steps S3121 to S3122, as follows: Step S3121: When the robotic arm performs the alignment action, determine the compensation distance of the pipette tip during alignment based on the oscillation radius R and oscillation angular velocity ω of the current horizontal oscillation mechanism. The compensation distance... The calculation formula is as follows: ; In the formula, This is the compensation distance for the pipette tip during alignment, so that the droplets sprayed from the pipette tip can land at the center of the corresponding orifice of the horizontal oscillation mechanism after falling. Let be the radius of rotation of the horizontal oscillating mechanism. The angular velocity of the horizontal oscillating mechanism. The flight time of a droplet from the tip of the pipette to its contact with the surface of the solid culture medium. This is a correction factor related to fluid viscosity and surface tension.

[0067] Specifically, The acquisition method is as follows: with the oscillation mechanism in a static state, a high-speed camera is mounted on the side of the pipette tip, the liquid discharge command is triggered, and the first frame image of the droplet leaving the tip of the pipette tip and the second frame image of the droplet contacting the surface of the culture medium are recorded.

[0068] Time of flight was measured when the oscillating mechanism was at rest. This is to eliminate interference from oscillations. Ignoring the effects of air eddies, horizontal oscillations do not change the vertical fall time of the droplet. By establishing baseline parameters under static conditions, the flight time can be... Defined as a constant physical constant of the system, thus simplifying subsequent calculation models in dynamic oscillations. Alternatively, the flight time can be measured when the oscillating mechanism is in a set operating condition. For example, the 800 rpm operating condition in this embodiment.

[0069] Specifically, The method for obtaining the data is as follows: Assuming the phase triggering accuracy of S313 meets the requirements, multiple spraying tests are conducted while the oscillation mechanism rotates at 800 rpm. The actual droplet landing point deviation on the culture medium surface, i.e., the distance between the actual landing point and the center of the well, can be measured using a high-speed camera. Since this embodiment sprays multiple droplets simultaneously onto the well, the center of the pipette tip can be aligned with the center of the well, and one droplet can be sprayed from the center of the pipette tip onto the well for easy observation. Assuming... The value was 1. After multiple experiments, based on the distance between the actual landing point and the center of the hole, the value was adjusted to 1. Adjustments are made to ensure that the actual landing point error of the droplet under high-speed rotation is within the preset error range.

[0070] Step S3122: Determine the preset coordinates of the pipette tip based on the compensation distance of the offset during alignment.

[0071] Specifically, the preset coordinates of the pipette tip are calculated using a formula. The addition and subtraction signs are selected according to the direction of rotation. In the formula, The center of the corresponding hole in the container. coordinate, Let Y be the center Y coordinate of the corresponding hole position in the container. This is the phase azimuth angle of the corresponding hole position in the container.

[0072] Specifically, in circular oscillation, each hole on the perforated plate has a fixed polar coordinate position relative to the rotation center of the oscillation platform. The coordinates of the corresponding hole position of the container on the perforated plate are given, which is the polar angle relative to the rotation center of the oscillating platform.

[0073] Because the hole center moves in a circle with the platform, its instantaneous direction of motion is always along the tangential direction of the circle, that is, perpendicular to the line connecting the hole center and the center of rotation. Therefore, the compensation distance... It needs to be decomposed along the tangential direction, that is, through the phase azimuth angle. Project it onto the X and Y axes of a Cartesian coordinate system.

[0074] By introducing the phase azimuth angle of the aperture position It can accurately calculate the components of the compensation distance on the X and Y axes, ensuring that the pipette tip can be precisely pre-biased in the tangential direction of the orifice's trajectory, regardless of the orifice's position on the oscillation platform. This compensates for the tangential displacement of the orifice's center during the droplet's flight time, ensuring that the droplet's final landing point can accurately fall into the orifice's geometric center under high-speed oscillation conditions.

[0075] The above technical solution effectively solves the problem of landing point deviation caused by the continuous movement of the culture medium surface under high-speed oscillation conditions by predicting the tangential displacement of the well center during the droplet flight time and pre-offsetting the alignment coordinates of the pipette tip. It ensures that the overlap error between the bacterial liquid landing point and the well center is controlled within the preset range, thereby improving the coating accuracy under high-frequency oscillation conditions and avoiding bacterial liquid splashing onto the well wall or cross-contamination between wells due to landing point deviation.

[0076] Step S313: While maintaining horizontal agitation on the surface of the solid culture medium, multiple pipette tips suspend the aspirated fluid and spray it onto the surface of the solid culture medium in the corresponding well.

[0077] Specifically, the horizontal oscillation mechanism operates in a circular oscillation mode. After the horizontal oscillation mixing module enters a stable, uniform circular motion, a suspended spraying operation is performed within a 0°±10° phase window of each oscillation cycle. The time for each drainage, i.e., the suspended spraying time, is ensured to be less than the phase window period. Suspended spraying begins when the container reaches 0°, thus guaranteeing the accuracy of the spray landing point and ensuring that the spray reaches the corresponding well positions. After completing the suspended spraying of each set of well positions, the horizontal oscillation mechanism can maintain oscillation for a few seconds to ensure uniform dispersion of the bacterial solution on the surface of the solid culture medium, and then it stops.

[0078] In some implementations, this embodiment divides the 24 holes into 6 groups of 4 holes each, and completes the full-board coating through step S313. Step S313 includes steps S3131 to S3134, and the specific steps are as follows: Step S3131: Obtain the angular velocity of the oscillation mechanism. Total time for pipette tip drainage .

[0079] Specifically, the total time for the pipette tip to discharge fluid is the sum of all the time elapsed from the moment the multichannel pipetting robot arm issues the discharge command until the moment the fluid actually begins to be discharged stably.

[0080] Specifically, the total time for the pipette tip to expel liquid. The control time is 15ms.

[0081] Step S3132: Obtain the current oscillation phase angle in real time. .

[0082] Step S3133: Based on angular velocity Total time for pipette tip to drain liquid and the current oscillation phase angle Determine the predicted phase angle of the oscillation mechanism when the pipette tip is discharging liquid. .

[0083] Specifically, the predicted phase angle is determined according to the formula. : ; In the formula, For the time elapsed since the current moment Subsequently, the phase angle that the oscillation mechanism is expected to reach, This represents the current oscillation phase angle of the oscillating mechanism. The angular velocity of the oscillating mechanism. This represents the total time for the pipette tip to expel liquid.

[0084] Step S3134: Based on the preset spray window, if If the spray is within the preset spray window, all channels will be simultaneously drained; if If it falls outside the window, it will wait to enter the next nearest safe window to trigger drainage.

[0085] Specifically, the current count value of the rotary encoder of the oscillation mechanism is obtained, and the current count value is converted into the current oscillation phase angle. Therefore, calculate according to the formula In this embodiment, the spraying window is a 0°±10° phase window for each oscillation cycle, once determined. If the temperature is within 0°±10°, it will trigger synchronous drainage from all channels. Outside the window, wait to enter the next nearest safe window to trigger drainage.

[0086] The above technical solution suffers from a delay between the issuance of the discharge command and the actual departure of the fluid from the pipette tip. If the discharge is triggered directly at the current phase, the orifice center will have already moved with the platform when the droplet is actually discharged. By introducing a predicted phase angle... The timing of the drainage command will be brought forward. · This ensures that the center of the orifice is exactly below the pipette tip the moment the droplet begins to leave the tip, providing the correct droplet landing point.

[0087] Steps S3131 to S3134 resolve the delay in response to the drainage command. The issue of mismatched droplet landing points is addressed by ensuring that the well center is precisely below the pipette tip when the droplet begins to leave the tip, providing the correct starting point for the droplet's fall—this is compensation in the time dimension. Steps S3121 to S3122 resolve the issue of tangential displacement of the well center caused by the continuous movement of the culture medium surface during the droplet's flight time τ after drainage. By pre-biasing the pipette tip's alignment coordinates, the final droplet landing point coincides with the well center after the displacement over time τ—this is compensation in the spatial dimension. The combined time and spatial compensation ensures that the droplet accurately falls into the well center at any rotational speed, avoiding landing point deviations and cross-contamination between wells caused by control delays or target displacement.

[0088] Step S400: After the multi-channel robotic arm completes the suspended spraying of fluid to a group of wells in the multi-well plate, the multi-channel robotic arm removes the currently gripped pipette tip and re-grips a new pipette tip.

[0089] By replacing the disposable pipette tip after each operation, the risk of cross-contamination between different wells or sample plates is completely avoided, ensuring the reliability of experimental results.

[0090] Step S500: The new pipette tip performs fluid suspension spraying on another set of wells in the multi-well plate.

[0091] Specifically, after completing the overhead spraying of this group of wells, the eight-channel robotic arm can move laterally by 19 mm to reach above the center of four wells in an adjacent column, in order to perform overhead spraying of the next group. Once the eight-channel robotic arm has picked up a new pipette tip and moved above the next group of wells, the horizontal oscillation mechanism can continue to operate.

[0092] Step S600: Repeatedly perform the operations of removing the suction head, grabbing the new suction head, and spraying in the air until the spraying in the air is completed for all holes.

[0093] In this embodiment, for a 24-well plate, a total of 6 cycles are required, with 4 wells sprayed each time, to complete the coating of all 24 wells.

[0094] Step S700: Cap the multiple wells and transfer the capped solid culture medium to an incubator for incubation.

[0095] After the entire plate is coated, the first robotic arm or the cap-grabbing robotic arm moves to the location where the cap was previously stored, grabs the cap, and accurately places it back onto the multi-well plate. This capping operation prevents contamination by other microorganisms and excessive water loss from the culture medium during incubation.

[0096] After the cover is applied, the first robotic arm picks up the multi-well plate from the horizontal oscillation mechanism and transfers it to an incubator or other designated incubation location. The incubator is set to a temperature suitable for microbial growth, and the incubation time is determined based on the microbial strain and experimental requirements.

[0097] Step S800: Transfer the source bacterial culture sample plate to the material transfer location for subsequent processing.

[0098] The second robotic arm picks up the sample container that has been sampled from the rack in the pipetting workstation and transfers it to the material transfer position. If there are other sample plates that need to be processed, the process from steps S120 to S700 is repeated; if all samples have been processed, the process ends and the workstation enters standby mode.

[0099] The following embodiments all adopt the following... Figure 2 The coating system shown performs... Figure 1 The methods shown in steps S100 to S800 are performed using an automated process for coating. The test bacterial solution is a standard E. coli culture, and the container is a 24-well plate, with each well containing a surface of antibiotic-treated solid culture medium. After coating, the 24-well plate is placed in a 37°C incubator and incubated for 18 to 24 hours. The colony distribution in each well is observed and recorded visually. The average colony count (CFU) and coefficient of variation are calculated, and the coating uniformity is rated as excellent, good, or poor. Excellent indicates uniform colony distribution with no aggregation or blank areas; good indicates relatively uniform colony distribution with a small amount of aggregation; and poor indicates severe colony aggregation or large areas of blank areas.

[0100] Other experimental instruments include: disposable sterile pipette tips, all of which are sterilized to avoid contamination; and an automated pipetting workstation compatible with Tecan Evo Freedom series pipetting workstations.

[0101] The test parameters were set as follows: This test adopted the single-factor variable method, setting variable gradients around four core parameters: spray volume, oscillation frequency, oscillation time, and suspension spray time point, while keeping the other parameters consistent. The specific variable gradients are as follows: (1) Spray volume: set to 50μL and 80μL respectively; (2) Oscillation frequency: set to 500 r / min, 800 r / min, and 1200 r / min respectively; (3) Oscillation time: set to 10s, 15s, and 20s respectively; (4) Suspended spray time points: set to before oscillation, during oscillation, and after oscillation respectively.

[0102] Example 1 1. Example 1 optimized the spraying time point. Under the conditions of fixed spray volume of 50μL or 80μL, oscillation frequency of 800r / min, and oscillation time of 15s, the coating effect was tested at three spraying time points: before oscillation, during oscillation, and after oscillation. Among them, "during oscillation" refers to the middle moment in the continuous oscillation process after the horizontal oscillation mechanism starts and reaches stable uniform oscillation.

[0103] Table 1 summarizes the coating effects at different spraying time points in Example 1; Table 1 Test number Spray volume (μL) Oscillation frequency (r / min) Oscillation time (s) Suspension spray timing Average colony count (CFU) Coefficient of variation (%) Coating uniformity (Excellent / Good / Poor) 1 50 800 15 Before the turmoil 878 17.2 good 2 50 800 15 During the fluctuations 673 8.2 excellent 3 50 800 15 After the shock 432 23.1 Difference 4 80 800 15 Before the turmoil 1022 14.5 good 5 80 800 15 During the fluctuations 809 10.1 excellent 6 80 800 15 After the shock 321 18.4 Difference 2. Analysis of the results in Example 1: First, the data in Table 1, showing the spraying time points during shaking (test numbers 2 and 5) and before shaking (test numbers 1 and 4), indicate that the coefficients of variation for spraying during shaking were 8.2% and 10.1%, respectively, lower than the 17.2% and 14.5% for spraying before shaking. This also shows that the coating uniformity improved from good to excellent. This indicates that during horizontal shaking, the droplets are uniformly dispersed and fixed by a stable tangential shear force the instant they contact the culture medium surface. This avoids the irregular displacement of static droplets caused by sudden acceleration changes at the start of subsequent shaking, as seen in spraying before shaking, thus effectively reducing the variability in colony distribution.

[0104] Second, comparing the data in Table 1 for the suspension spraying time points during shaking (test numbers 2 and 5) and after shaking (test numbers 3 and 6), it can be seen that the coefficients of variation for suspension spraying after shaking are as high as 23.1% and 18.4%, respectively, and the coating uniformity is poor in both cases. This is because when suspension spraying is performed after shaking stops, the droplets, after being added to the static culture medium surface, lose the continuous effect of tangential shear force. The evaporation rate at the edge of the droplet is higher than that at the center, causing bacteria to migrate to the edge and form ring-shaped deposits at the edge, resulting in severe colony aggregation. However, when suspension spraying is performed during shaking, the tangential shear force generated by horizontal shaking continuously acts on the inside of the droplet, preventing the bacteria from forming stable edge aggregation, thus obtaining uniformly distributed isolated monoclonal colonies.

[0105] Third, comparing test numbers 2 (spray volume 50μL) and 5 (spray volume 80μL), it can be seen that under the same oscillation frequency and oscillation time, the coefficient of variation of the 50μL spray volume is 8.2%, which is lower than the 10.1% of the 80μL. This indicates that the 50μL spray volume is preferred, as it can appropriately reduce the spray volume and is beneficial to further improve the coating uniformity.

[0106] In summary, the optimal spraying time is one that fully utilizes the tangential shear force generated during oscillation, dispersing and fixing the droplets the instant they contact the culture medium surface. This avoids irregular droplet displacement caused by sudden acceleration changes during oscillation start-up or cessation, and is crucial for achieving highly uniform coating. Furthermore, the optimal spray volume is 50 μL.

[0107] Example 2 1. Example 2 optimized the oscillation frequency and oscillation time. Under the condition of a fixed spray volume of 50 μL and a spraying time point during oscillation, the effects of different combinations of oscillation frequencies (500 r / min, 800 r / min, 1200 r / min) and oscillation times (10 s, 20 s) on the coating effect were investigated. Each set of conditions was repeated 3 times.

[0108] Table 2 summarizes the coating effects of different combinations of oscillation frequencies and oscillation times in Example 2; Table 2 Test number Spray volume (μL) Oscillation frequency (r / min) Oscillation time (s) Suspension spray timing Average colony count (CFU) Coefficient of variation (%) Coating uniformity (Excellent / Good / Poor) 1 50 500 10 During the fluctuations 324 11.2 good 2 50 800 10 During the fluctuations 378 9.8 good 3 50 1200 10 During the fluctuations 563 7.6 excellent 4 50 500 20 During the fluctuations 377 11.4 good 5 50 800 20 During the fluctuations 491 10.4 good 6 50 1200 20 During the fluctuations 663 6.5 excellent 2. Analysis of the results in Example 2: First, comparing data from different oscillation frequencies under the same oscillation time, it can be seen that when the oscillation time is 10s, as the oscillation frequency increases from 500r / min to 1200r / min, the coefficient of variation gradually decreases from 11.2% to 7.6%, and the coating uniformity improves from good to excellent. When the oscillation time is 20s, as the oscillation frequency increases from 500r / min to 1200r / min, the coefficient of variation decreases from 11.4% to 6.5%, and the coating uniformity also improves from good to excellent. This indicates that a higher oscillation frequency can provide stronger tangential shear force, allowing the bacterial solution to be uniformly dispersed into smaller units in a shorter time, which helps to form more isolated monoclonal colonies, while reducing the variability of colony distribution.

[0109] Second, comparing data from different shaking times at the same shaking frequency reveals that, when the shaking frequency is consistently 1200 r / min, extending the shaking time from 10 s to 20 s further reduces the coefficient of variation from 7.6% to 6.5%, while increasing the average colony count from 563 CFU to 663 CFU. This indicates that appropriately extending the shaking time helps the bacterial solution to spread and dry fully under horizontal shear force, further reducing the variability in colony distribution and improving the uniformity of monoclonal colonies.

[0110] Third, test number 6 (oscillation frequency 1200 r / min, oscillation time 20 s) achieved the best coating effect, with a coefficient of variation as low as 6.5% and excellent coating uniformity. This indicates that under this parameter combination, the bacterial solution can be fully dispersed and evenly attached to the surface of the culture medium under horizontal oscillation, and the single clonal colony distribution is the most uniform and the isolation is the best.

[0111] In summary, appropriately increasing the oscillation frequency and extending the oscillation time both enhance the shear dispersion effect, thereby improving the coating uniformity. Under the conditions of this embodiment, the optimal oscillation parameters are: oscillation frequency 1200 r / min and oscillation time 20 s.

[0112] Through steps S100 to S800, this embodiment achieves fully automated, high-throughput bacterial culture coating, from material transfer, shaking start-up, suspended spraying, pipette tip replacement to capping and incubation. This method eliminates the need for contact coating tools such as coating sticks and glass beads. During the coating process, the pipette tip never comes into contact with the culture medium surface, avoiding problems such as culture medium puncture, cross-contamination, and difficulties in sterilizing tools. Simultaneously, the synergistic effect of horizontal shaking and suspended spraying improves the uniformity and isolation of single-clone colonies.

[0113] The beneficial effects of this application are: 1. In this application, the suspension spraying and oscillation are carried out simultaneously. When the droplet contacts the surface of the culture medium, the horizontal oscillation mechanism is already in uniform periodic motion and there is no sudden acceleration. Therefore, the droplet is fixed near the landing point at the moment of contact and will not undergo large displacement.

[0114] 2. In this application, when the droplet contacts the surface of the culture medium, the horizontal oscillation mechanism is already in uniform periodic motion. The tangential shear force generated by the oscillation continuously acts on the inside of the droplet, preventing the bacteria from forming stable edge aggregations and deposits. Instead, the bacteria are uniformly dispersed and randomly fixed at various positions within the droplet's coverage area. Compared to a statically added droplet, the evaporation rate at the droplet edge is higher than at the center, causing the suspended bacteria to be transported from the center to the edge. This results in ring-shaped deposits at the edge, similar to coffee rings, which severely affects the isolation of monoclonal colonies.

[0115] 3. In this application, steps 3121-S3122 and steps 3131-S3134 ensure that the droplets can fall into the preset positions of the corresponding wells, thereby improving the uniformity of the distribution of monoclonal colonies and avoiding the risk of cross-contamination.

[0116] 4. Under the conditions of the embodiments of this application, the optimal combination of automated coating parameters is: spray volume 50 μL, oscillation frequency 1200 r / min, oscillation time 20 s, and the spraying time point is during oscillation. Under this parameter combination, the coefficient of variation is as low as 6.5%, the coating uniformity reaches the excellent level, the single clonal colony distribution is uniform and the isolation is good, which fully meets the process requirements of high-throughput automated bacterial solution coating.

[0117] Please see Figure 2 , Figure 2 This is a schematic diagram of the coating system provided in an embodiment of this application. Figure 2As shown, the coating system includes: a control unit, a horizontal oscillation mechanism, a pipetting device, a first robotic arm, and a second robotic arm.

[0118] A control unit can be a computer system consisting of one or more processors and memory. The memory stores executable program instructions, and the processor executes these instructions to control the actions of various mechanisms.

[0119] The control unit is communicatively connected to the horizontal oscillation mechanism, the pipetting device, the first robotic arm, and the second robotic arm to control the actions of each mechanism: controlling the first robotic arm to transfer the container from the first position to the horizontal oscillation mechanism; controlling the second robotic arm to transfer the sample container carrying fluid from the second position to the reach of the pipetting device so that the pipetting device can draw fluid from the sample container; controlling the horizontal oscillation mechanism to start so that the surface of the solid culture medium is in a horizontal oscillation state; and controlling the pipetting device to spray the fluid to be coated onto the surface of the solid culture medium while maintaining horizontal oscillation on the surface of the solid culture medium.

[0120] The horizontal oscillation mechanism is used to support containers with a solid culture medium surface inside and to apply horizontal oscillation motion to the containers. Specifically, the horizontal oscillation mechanism includes an oscillation platform, a drive motor, and a clamping component. The oscillation platform is used to hold the container, and the clamping component is located at the edge of the oscillation platform to clamp and fix the container's edge after placement, preventing displacement of the container during oscillation. The drive motor is connected to the oscillation platform and drives the oscillation platform to perform periodic movements in the horizontal plane under the command of the control unit.

[0121] A pipetting device is used to aspirate fluid for coating and to spray the fluid onto the surface of a solid culture medium. Preferably, the pipetting device is a multi-channel pipetting robot arm, such as an eight-channel pipetting robot arm. Specifically, the multi-channel pipetting robot arm includes multiple connected pipetting channels and motion components. Each pipetting channel includes a pipetting pump, and its end is used to load replaceable pipetting tips. The pipetting pump is used to aspirate and dispense the liquid, and the motion components include X-axis, Y-axis, and Z-axis motion mechanisms for driving the pipetting tips to move in three-dimensional space.

[0122] Both the first and second robotic arms consist of a robotic arm body and grippers. The robotic arm body is a multi-axis articulated or Cartesian coordinate motion mechanism, which can move flexibly within the workspace. The grippers are mounted at the end of the robotic arm body, and their shape and size are adapted to the edge of the container or sample container for stable gripping and releasing of the container or sample container.

[0123] The workflow of the coating system in this application embodiment is as follows: Step 1: Control unit: Sends instructions to the first robotic arm to transfer the container carrying the solid culture medium surface from the first position to the oscillation platform of the horizontal oscillation mechanism, and locks the container with a clamping device.

[0124] Step 2: The control unit sends a command to the second robotic arm to transfer the sample container containing the bacterial solution to be coated from the second position to the sample carrier within the reach of the pipetting device.

[0125] Step 3: The control unit starts the horizontal oscillation mechanism, and the oscillation platform drives the container to oscillate horizontally, so that the surface of the solid culture medium enters a stable horizontal oscillation state.

[0126] Step 4: While the horizontal oscillation continues, the control unit controls the pipetting device to grab the pipette tip, draw a preset volume of bacterial solution from the sample container, move it above the first set of wells in the container, and apply the bacterial solution to the surface of the solid culture medium by means of suspended spraying.

[0127] Step 5: After the suspension spraying is completed, the control unit controls the multi-channel pipetting robot to remove the used pipette tip, pick up the pipetting tip again, and perform suspension spraying on the next set of wells. The control unit can also control the horizontal oscillation mechanism to continue running for a preset time to ensure that the bacterial solution is fully spread and initially dried.

[0128] Step 6: After coating is completed, the first robotic arm will cover the container and transfer it to a designated location, such as an incubator, for incubation.

[0129] In summary, this application provides a coating method and coating system. The coating method includes providing a container carrying a solid culture medium surface; applying horizontal agitation to the container to keep the solid culture medium surface in a horizontal agitation state; while the solid culture medium surface is kept in a horizontal agitation state, spraying a fluid to be coated, which is a bacterial suspension containing microorganisms, onto the solid culture medium surface; wherein, when the fluid comes into contact with the horizontally agitated solid culture medium surface, it is dispersed by the horizontal agitation and adheres to different areas of the solid culture medium surface due to adhesive forces to form isolated monoclonal colonies. This application can simultaneously meet the requirements of high-throughput automation adaptability and uniform distribution of monoclonal colonies, and further improves the monoclonal colony acquisition rate.

[0130] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of this application, and are not intended to limit them. Although this application has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of this application.

Claims

1. A coating method, characterized in that, The coating method includes: Provide a container that carries the surface of a solid culture medium; The container is subjected to horizontal agitation to bring the surface of the solid culture medium into a horizontal agitation state. While maintaining horizontal agitation on the surface of the solid culture medium, the fluid to be coated is sprayed onto the surface of the solid culture medium in mid-air, the fluid being a bacterial solution containing microorganisms; When the fluid comes into contact with the surface of the solid culture medium, which is in a horizontal oscillation state, it is dispersed by the horizontal oscillation and adheres to different areas of the solid culture medium surface due to adhesive force, so as to form isolated monoclonal colonies. After providing a container carrying the surface of a solid culture medium, the coating method further includes: The container is transferred from a first position to the horizontal oscillation mechanism by a first robotic arm, the horizontal oscillation mechanism being used to apply horizontal oscillation to the container; The sample container containing the fluid is transferred from the second position to the reach of the pipetting device by the second robotic arm, so that the pipetting device can draw the fluid from the sample container. The pipetting device is used to draw the fluid to be coated and spray the fluid to be coated onto the surface of the solid culture medium.

2. The coating method according to claim 1, characterized in that, The coating method further includes: The fluid is sprayed onto the surface of the solid culture medium in the form of multiple droplets, and each droplet experiences a greater adhesive force than centrifugal force under horizontal oscillation, thus adhering to the vicinity of the spraying location.

3. The coating method according to claim 1, characterized in that, The container is a multi-well plate, which includes multiple wells, each of which carries the surface of the solid culture medium. While maintaining horizontal agitation on the surface of the solid culture medium, the fluid to be coated is sprayed onto the surface of the solid culture medium in a suspended state, including: While maintaining horizontal agitation on the surface of the solid culture medium, the fluid is simultaneously or sequentially sprayed into the surface of the solid culture medium at multiple wells of the multi-well plate using a pipetting device.

4. The coating method according to claim 3, characterized in that, The pipetting device includes a multi-channel robotic arm, which, while maintaining horizontal oscillation on the surface of the solid culture medium, suspends and sprays the fluid to be coated onto the surface of the solid culture medium, including: The multi-channel robotic arm grasps a corresponding number of pipette tips so that the spacing between the pipette tips matches the pore spacing of the multi-well plate, and the multiple pipette tips aspirate the fluid from the sample container; Move the plurality of pipette tips above the corresponding multiwell plates; While maintaining horizontal agitation on the surface of the solid culture medium, the multiple pipette tips suspend the aspirated fluid and spray it onto the surface of the solid culture medium in the corresponding well.

5. The coating method according to claim 1, characterized in that, The volume of fluid sprayed onto the surface of the solid culture medium is 30 to 80 μL.

6. The coating method according to claim 4, characterized in that, Before the plurality of pipette tips aspirate the fluid from the sample container, the method further includes: The fluid in the sample container is mixed by blowing and sucking multiple times.

7. The coating method according to claim 4, characterized in that, The method also includes: After the multi-channel robotic arm completes the suspended spraying of fluid to a group of orifices in the multi-hole plate, the multi-channel robotic arm removes the currently gripped pipette tip and re-grips a new pipette tip. The new pipette tip performs fluid suspension spraying on another set of wells in the multi-well plate; Repeat the process of removing the suction head, grabbing the new suction head, and spraying in the air until all holes have been sprayed in the air. The multiple wells are capped, and the capped solid culture medium is transferred to an incubator for incubation.

8. A coating system, characterized in that, include: A horizontal oscillation mechanism is used to support a container with a solid culture medium surface and to apply horizontal oscillation to the container; A pipetting device is used to pick up the fluid to be coated and spray the fluid onto the surface of the solid culture medium while suspended in the air. The control unit is communicatively connected to the horizontal oscillation mechanism and the pipetting device, and is used to control the activation of the horizontal oscillation mechanism so that the surface of the solid culture medium is in a horizontal oscillation state. And while maintaining horizontal oscillation on the surface of the solid culture medium, the pipetting device is controlled to spray the fluid to be coated onto the surface of the solid culture medium from a suspended position.

9. The coating system according to claim 8, characterized in that, It also includes a first robotic arm and a second robotic arm, both of which are communicatively connected to the control unit. The first robotic arm is used to transfer the container from the first position onto the horizontal oscillation mechanism; The second robotic arm is used to transfer a sample container containing the fluid from a second position to within the reach of a pipetting device, so that the pipetting device can draw the fluid from the sample container.

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