Imprinted bacterial suspension array inoculation block, device and method

Abstract

This invention belongs to the technical field of bacterial culture transfer equipment, and more specifically, relates to an imprinted bacterial culture dot matrix inoculation block, device, and method. The invention includes a thin-film inoculation block body, which is divided into a smooth pickup section and an imprinted inoculation section. The imprinted inoculation section includes at least one pressure relief groove and dot matrix protrusions located on both sides of the pressure relief groove. Each dot matrix protrusion includes several raised portions, which are symmetrically and evenly distributed in a matrix on both sides of the pressure relief groove. Each raised portion has a concentric recess at its top center. This invention, through its thin-film inoculation block body, through-type pressure relief groove, matrix dot matrix protrusions, and concentric recess structure, combined with a porous liquid storage pad for quantitative liquid supply and an automated pickup and imprinting mechanism, solves the problems of poor consistency, vacuum adhesion, cross-contamination, low automation, and lack of process traceability in existing inoculation methods, achieving high-throughput, standardized, pollution-free, and traceable bacterial culture dot matrix inoculation.

Description

Technical Field

[0001] This invention belongs to the technical field of bacterial culture transfer equipment, and more specifically, relates to an imprinted bacterial culture dot matrix inoculation block, device, and method. Background Technology

[0002] In bacterial culture, colony counting, clone screening, antimicrobial susceptibility testing, and optimization of culture conditions, it is often necessary to inoculate bacterial suspensions onto the surface of solid agar culture dishes. Traditional methods include swab application, streaking with an inoculation loop, spreading with a spreader, drop plate method, replication plate, and multi-needle array transfer. These methods each have their uses, but in automated batch inoculation, there are still problems with consistency, cycle time, contamination control, and pattern control. Manual swabs or spreaders are suitable for forming bacterial mosses or spreading over large areas, but the amount of liquid absorbed, the spreading pressure, and the trajectory depend on the operator; multi-needle arrays or replication plates are suitable for dot-matrix transfer from existing colonies or bacterial libraries, but the bacteria are usually carried directly by the needle tip, cloth, or impression, making it difficult to stably handle quantitative supply of a single liquid bacterial sample, prevent dripping, and perform regular dot-matrix imprinting; pipetting allows for the addition of bacterial suspension according to coordinates, but the spread, drying, and absorption of the droplets are greatly affected by the surface condition of the culture dish.

[0003] Chinese patent document CN206157135U discloses a rectangular sample loading platform for a multi-point inoculation instrument and a matching rectangular culture dish. The vertical tube and locking structure reduce bacterial liquid dripping, and the liquid level plate controls the liquid level height. Although it improves inoculation efficiency and consistency, it still has problems such as the inoculation head and culture dish easily forming a vacuum adhesion, dragging agar when lifted, and not being compatible with the automated feeding of thin-film disposable inoculation blocks.

[0004] Chinese patent document CN105255715A discloses a high-throughput microbial inoculation device and a method for batch inoculating bacterial strains. It uses an array of inoculation needles and a sample tube rack to achieve batch dipping and synchronous inoculation. Although it improves screening throughput and comparison consistency, it still has problems such as unstable quantitative supply of liquid bacterial solution, easy cross-contamination of needles, inability to achieve imprint-type dot matrix transfer and full-process data traceability.

[0005] In summary, there is an urgent need to provide an imprinted bacterial liquid array inoculation block, inoculation device, and inoculation method to solve the problems of poor inoculation consistency, vacuum adhesion, high risk of contamination, low degree of automation, and lack of process traceability in the existing technology. Summary of the Invention

[0006] The technical problem to be solved by this invention is to address the issues of poor consistency, vacuum adhesion, cross-contamination, low automation, and lack of process traceability in existing inoculation methods by using a thin-film inoculation block body, a through-type pressure relief groove, a matrix-type dot matrix protrusion and concentric depression structure, combined with a porous liquid storage pad for quantitative liquid supply and an automated pick-up and imprinting mechanism. This results in high-throughput, standardized, pollution-free, and traceable bacterial dot matrix inoculation.

[0007] The technical terms related to this invention are explained as follows: Imprint inoculation: The bacterial solution carried by the protrusions is transferred to the surface of the petri dish by contact pressure to form a regular dot matrix.

[0008] Pressure relief groove: A groove that runs through the lower surface and periphery of the inoculation block body, used to break the vacuum negative pressure when it is lifted.

[0009] Matrix protrusions: A matrix-distributed protrusion structure used to support and transfer bacterial solution.

[0010] Recessed portion: A microstructure located at the top of the protrusion, used for capillary dipping of a quantitative bacterial solution.

[0011] Smooth pickup section: A smooth area on the upper surface of the inoculation block body, used for stable pickup by the negative pressure nozzle.

[0012] Porous liquid storage pad: A liquid supply carrier made of porous material, used to uniformly carry and stably release bacterial liquid.

[0013] Hydrophilic micro-roughened surface: The micro-roughened structure formed by surface treatment improves capillary adhesion stability.

[0014] Annular guard: A raised structure located on the outer periphery of the inoculation part to prevent accidental contact with non-working surfaces.

[0015] Gradient dilution: The bacterial culture is prepared in multiple concentration levels to meet different experimental needs.

[0016] Vision and Traceability: Through image acquisition and data recording, the entire vaccination process can be traced.

[0017] The technical problem to be solved by this invention is achieved by the following technical solution: An imprinted bacterial suspension array inoculation block includes a thin-film inoculation block body, which is divided into the following parts: A smooth pickup section is located on the upper surface of the inoculation block body and has a smooth pickup area; The imprinting inoculation section, located on the lower surface of the inoculation block body, includes at least one pressure relief groove and dot matrix protrusions located on both sides of the pressure relief groove, wherein: The pressure relief channel extends from the center of the lower surface to both sides of the periphery of the lower surface to form a through opening. The dot matrix protrusions include several protrusions that are symmetrically and evenly distributed in a matrix on both sides of the pressure relief channel. Each protrusion has a concentric recess at the center of its top.

[0018] This technical solution achieves regular inoculation points, uniform pick-up volume, no vacuum adhesion when lifted, and no dripping when moved by functional partitioning of upper and lower surfaces, through a through-type pressure relief structure, a matrix-style protrusion layout, and a capillary structure of concentric recesses. The thin-sheet body is adapted for automated pickup and single use, reducing the risk of cross-contamination and improving inoculation consistency and throughput.

[0019] Preferably, in this invention, the inoculation block body is a thin-film structure adapted to the shape of a petri dish, and the depth of the pressure relief groove is less than the height of the dot matrix protrusions. This technical solution, through the limitation of shape and height difference, ensures structural strength, pressure relief effectiveness, imprinting stability, and capillary pick-up accuracy, and is compatible with automated pickup and imprinting control parameters.

[0020] Preferably, the pickup area of ​​this invention is a flat surface, a shallow concave surface, or a mating surface that adapts to the sealing ring. The pickup area is provided with a QR code, color mark, batch identification code, asymmetric error-proofing notch, or a single-sided chamfer. This technical solution improves the stability of negative pressure mating through diversified pickup surface structures, reduces assembly error rate through identification and error-proofing structures, and realizes batch management and posture recognition.

[0021] Preferably, the pressure relief groove of this invention is one or more combinations of straight grooves, zigzag grooves, curved grooves, intersecting grooves, or radial grooves. The through opening is a straight opening, an arc-shaped opening, or a geometrically regular opening, extending through to the side wall of the inoculation block body. The pressure relief groove is located between the lattice protrusions but avoids the inoculation position of the lattice protrusions, and the depth of the pressure relief groove is greater than the depth of the recess. This technical solution ensures unobstructed air passage through the multi-form groove and opening layout, avoids affecting the inoculation point through positional avoidance, and ensures pressure relief effect through depth difference.

[0022] Preferably, the protrusions of this invention are cylindrical, frustum-shaped, or hemispherical structures, while the recesses are one or more combinations of micro-blind holes, micro-through holes, annular microgrooves, cross-shaped microgrooves, or radial microgrooves. This technical solution improves versatility by adapting to different bacterial solutions viscosities, pick-up volumes, and transfer effects through multi-shaped protrusions and recesses.

[0023] Preferably, the dot-matrix protrusions of this invention have hydrophilic micro-roughened surfaces, which are formed on the end faces of the protrusions and the inner walls of the recesses. These hydrophilic micro-roughened surfaces are one or more of the following: frosted surface, sandblasted surface, laser-etched surface, chemically etched surface, plasma-treated surface, or UV-ozone-treated surface. This technical solution enhances capillary action through the hydrophilic micro-roughened structure, stabilizes the pick-up amount, and reduces residue and dripping.

[0024] Preferably, the outer periphery of the imprinting inoculation section of this invention is provided with an annular guard, which surrounds the dot matrix protrusions. The height of the annular guard is lower than the height of the protrusions, and a height difference is formed between the bottom end face of the annular guard and the recessed part of the protrusion. This technical solution uses the guard to prevent non-working surfaces from contacting the culture dish or liquid storage pad, thus preventing contamination and liquid stain diffusion.

[0025] The present invention also provides an imprint-type bacterial culture array inoculation device.

[0026] An imprinted bacterial suspension array inoculation device, employing the aforementioned imprinted bacterial suspension array inoculation block, the device comprising: The supply mechanism is used to supply the inoculation block body, including a clip-type, drawer-tray type, stacked hopper type or rotary table type supply unit, and the supply unit is arranged adjacent to the working position of the picking mechanism. The pickup mechanism is used to pick up the smooth pickup part by negative pressure. It includes a negative pressure nozzle, a vision / negative pressure / height detection unit and an anti-inversion recognition unit. The negative pressure nozzle moves above the supply mechanism, the liquid supply mechanism and the culture dish support module. The liquid supply mechanism is a porous liquid storage pad, and each porous liquid storage pad is filled with bacterial liquid. The liquid supply mechanism is located below the moving path of the picking mechanism. The dilution mechanism is used to perform gradient dilution of the bacterial solution from the liquid supply mechanism, and is connected to the liquid supply mechanism. The liquid replenishment / pad replacement mechanism is used to replenish / replace the liquid supply mechanism. It is set up adjacent to the liquid supply mechanism and is triggered by the number of dips, weight signal changes, or image signals to replenish / replace the liquid. The disposal / recycling mechanism is used to dispose of / recycle the inoculation block body after inoculation, including a biohazardous waste bin / offline sterilization and recycling bin, which is located at the end of the movement path of the picking mechanism; A visual and traceability system is used to record relevant information about the inoculation process, including batch number, concentration, imprint location, spot diameter, and petri dish number. The control mechanism is electrically connected to the pickup mechanism, liquid supply mechanism, dilution mechanism, liquid replenishment / pad replacement mechanism, waste / recycling mechanism, and vision and traceability mechanism, respectively, and is used to control the imprinting pressure, position, dwell time, and number of repetitions of the imprinting inoculation section, and coordinate the timing of actions.

[0027] This technical solution achieves full automation of the inoculation process through modular integration and time-series collaboration, including material feeding, picking, liquid supply, dipping, imprinting, traceability, and recycling, thereby improving inoculation efficiency and stability.

[0028] Preferably, the porous liquid storage pad in the liquid supply mechanism of this invention is one or more of the following: sterile sponge, PVA sponge, non-woven fabric, cellulose pad, microporous polyurethane pad, microporous silicone pad, or sintered porous plastic pad. The porous liquid storage pad is configured with a strip-type, matrix-type, ring-type, or multi-compartment tray-type partitioned structure. The overall pore size of the porous liquid storage pad is 10μm-2mm, the overall thickness is 0.5mm-20mm, the pre-added liquid volume is 10μL-10mL, the contact pressure of the porous liquid storage pad is 0.01N-10N, and the contact time is 0.1s-10s. This technical solution, through optimization of materials, structure, and parameters, ensures uniform distribution and stable release of the bacterial solution, and is suitable for gradient experiments and parallel processing of multiple samples.

[0029] The present invention also provides a method for inoculating bacterial suspensions using an imprinting method.

[0030] An imprint-type bacterial suspension array inoculation method includes the following steps: S1. Supplying inoculation blocks: The sterilized inoculation block body is loaded into the supply mechanism and supplied in the form of a clip, drawer tray, stacked hopper or turntable. S2. Preparation of the liquid supply pad: Place the porous liquid storage pad into the liquid supply mechanism and add a predetermined volume of bacterial solution into the porous liquid storage pad; S3, Negative Pressure Pickup: The inoculation block body is picked up from the supply mechanism through the negative pressure nozzle of the pickup mechanism, and the correct posture is confirmed by vision, negative pressure or foolproof structure. S4. Capillary dipping: The picking mechanism drives the inoculation block body to descend, so that the dot matrix protrusions of the imprinted inoculation part contact the porous liquid storage pad with a predetermined pressure, and pick up the bacterial liquid through capillary action. S5. Moving and positioning: The picking mechanism moves the inoculation block body to the culture dish; S6, Dot Imprinting: The control mechanism drives the inoculation block body to press down, so that the dot matrix protrusions contact the culture dish with a predetermined pressure and residence time, and the bacterial solution is transferred to form a regular colony dot matrix; at the same time, the pressure relief channel maintains air communication through the through opening, breaking the negative pressure between the bottom surface and the culture dish. S7. Static absorption: After imprinting, allow the inoculum block to stand for a short time to allow the bacterial solution to be fully absorbed by the culture medium. S8. Image traceability: Detects the position, integrity, liquid spot diameter, and presence of continuous liquid droplets of the imprinted dot matrix through vision and traceability mechanisms; S9. Discard / Recycle: The picking mechanism releases the used inoculation block body to the discard / recycle mechanism, then picks up the next inoculation block body and enters the next cycle.

[0031] This technical solution constructs a closed-loop logic across the entire chain of "supply-dipping-transfer-verification-circulation," achieving continuous and standardized supply of the inoculum block body through a modular, multi-form supply architecture (clamp / tray / hopper / rotary). It also utilizes a triple verification mechanism of negative pressure suction and visual / negative pressure / mistake-proofing to ensure the correctness of the picking posture and operational reliability in each step. Furthermore, it replaces traditional active pipetting with capillary action, allowing the dot matrix protrusions of the imprint inoculum to uniformly pick up a predetermined volume of bacterial solution through passive wetting, eliminating pipetting errors and cross-contamination risks at the physical mechanism level. In the transfer stage, precise control of the pressing pressure and residence time achieves regular dot matrix transfer of the bacterial solution to the culture dish. Simultaneously, it introduces a pressure relief channel and an air-connected structure with a through-opening to break the negative pressure formed between the bottom surface and the culture medium at the moment of imprinting. This method fundamentally solves the problems of colony stringing, deformation, and incomplete transfer caused by negative pressure adsorption in imprint inoculation. After imprinting, a brief static setting promotes the full absorption and solidification of the bacterial solution by the culture medium. Then, an image traceability mechanism performs multi-dimensional quality detection on the dot matrix position, integrity, liquid spot diameter, and contiguous droplets, forming an "operation-feedback" quality closed loop. Finally, a pick-up-release-discard / recycling cycle mechanism drives the next round of inoculation, ensuring that the entire method achieves controllability, traceability, and repeatability of each step while maintaining high throughput. Essentially, it reconstructs the traditional serial operation of point-by-point inoculation into a parallel batch processing logic based on physical contact transfer, with negative pressure management and capillary dipping as the two core physical mechanisms throughout, thereby achieving the goal of high-precision, high-throughput, and fully traceable automated bacterial dot matrix inoculation.

[0032] The one or more technical solutions provided by this invention have the following advantages compared with the prior art: (1) High inoculation consistency, the dot matrix protrusions and concentric depressions ensure stable position, spacing and amount of saturation, the porous liquid storage pad provides uniform liquid supply, and the regular colonies facilitate image recognition and high-throughput screening.

[0033] (2) Stable and reliable operation, through-type pressure relief groove breaks vacuum adhesion, the inoculation block is lifted without dragging, the imprinting parameters are controllable, and it is suitable for automated continuous operation.

[0034] (3) Strict pollution control, disposable or offline sterilized inoculation blocks avoid cross-contamination, ring-shaped baffles prevent accidental contact, and the liquid supply and inoculation zones are independent, which improves the safety of the test. Attached Figure Description

[0035] Figure 1 This is one of the structural schematic diagrams of the inoculation block of the present invention.

[0036] Figure 2 This is the second schematic diagram of the structure of the inoculation block of the present invention.

[0037] Figure 3 This is a diagram showing the usage status of the inoculation device of the present invention.

[0038] In the figure: 1. Inoculation block body; 2. Pressure relief groove; 3. Dot matrix protrusion; 31. Protrusion; 32. Recess; 4. Edge guard; 5. Pick-up mechanism; 6. Porous liquid storage pad; 7. Bacterial solution. Detailed Implementation

[0039] Example 1 Please refer to Figure 1 , Figure 2 This embodiment provides an imprinted bacterial culture array inoculation block.

[0040] The inoculation block body 1 adopts an integrated molding structure, and its overall shape can be adapted to the shape of the petri dish. It is preferably designed as a rectangular thin sheet structure, with a regular shape that facilitates automated feeding and stacking. All four corners are rounded to avoid scratches, jamming, and stress concentration problems caused by sharp edges during handling, picking, and imprinting operations, effectively improving operational safety and overall structural durability. The aspect ratio of the inoculation block body 1 is preferably set at 2:1, and the thickness is preferably one-tenth of its length, with the main body thickness controlled within the range of 1mm-4mm. This thin sheet structure can ensure sufficient structural rigidity, adapt to various automated feeding modes such as clips, hoppers, and pallets, and also reduce raw material consumption and lower overall manufacturing costs.

[0041] The entire upper surface of the inoculation block body 1 is designed as a smooth pickup area. The pickup area surface remains flat and smooth, without protrusions, burrs, or grooves, enabling a stable and reliable seal with the negative pressure nozzle of automated equipment, ensuring that the inoculation block does not fall off, misalign, or tilt during the pickup process. The pickup area surface can be equipped with asymmetrical anti-foolproof notches and QR code markings according to usage requirements. The asymmetrical anti-foolproof notches ensure that the inoculation block maintains a unique and correct posture during supply and pickup, avoiding reverse, deflection, or misalignment pickup. The QR code markings can record production batch, material information, sterilization status, and usage restrictions, enabling full lifecycle traceability management.

[0042] The entire lower surface of the inoculation block body 1 is designed as an imprinting inoculation section, which is the core area for bacterial suspension dipping, transfer, and pressure relief. A pressure relief groove 2 is formed in the center of the imprinting inoculation section, extending from the central area of ​​the lower surface to the periphery on both sides, thus forming a through opening. This through opening directly penetrates the sidewall of the inoculation block body 1, ensuring continuous communication between the inside of the pressure relief groove 2 and the external atmosphere, forming a stable and unobstructed air channel. This air channel rapidly introduces outside air when the imprinting is completed and the inoculation block is lifted upwards, completely eliminating the negative pressure adsorption formed between the lower surface and the culture medium due to the sealed compression, preventing the agar layer from being dragged, torn, or deformed, and ensuring the integrity of the lattice morphology.

[0043] In this embodiment, the pressure relief channel 2 has a depth of 0.3 mm and a width of 0.5 mm. The depth of the channel is strictly less than the overall height of the protrusion 31, ensuring that the pressure relief channel 2 will not directly contact the porous liquid storage pad 6 during the dipping and imprinting process, and will not interfere with the normal dipping and transfer of the bacterial solution 7. The pressure relief channel 2 is located between the dot matrix protrusions formed by the dot matrix protrusions 3, with a central position that completely avoids the inoculation points corresponding to all the dot matrix protrusions, so as not to destroy the continuity and uniformity of the protrusion structure, while ensuring the shortest pressure relief path and the highest pressure relief efficiency.

[0044] The pressure relief channel 2 has symmetrically arranged dot matrix protrusions 3 on both sides. All protrusions 31 are evenly and equidistantly distributed in a matrix, with regular arrangement and consistent spacing, ensuring that the colony matrix formed after imprinting is neat and standardized, facilitating subsequent image recognition, colony counting, and data analysis. In this embodiment, the protrusions 31 adopt a cylindrical structure, which is symmetrical, has uniform force, and stable transfer. The diameter is uniformly 0.8 mm, and the height is uniformly 0.2 mm. The center-to-center distance between adjacent protrusions is set to 2 mm. The reasonable spacing can prevent the bacterial liquid from spreading and merging together, ensuring that each point grows independently.

[0045] Each protrusion 31 has a concentric recess 32 at the center of its top. The recess 32 is coaxial with the protrusion. In this embodiment, the recess 32 adopts a micro-blind hole structure with a pore diameter of 0.1 mm and a depth of 0.05 mm. The micro-blind hole structure can stably adsorb trace amounts of bacterial liquid using capillary action. It does not drip or flow during movement and can release a quantitative amount during imprinting, thus achieving controllable micro-transfer.

[0046] The top end face of the protrusion 31 and the inner wall surface of the depression 32 are uniformly treated with plasma to form a uniform and fine hydrophilic micro-rough surface. The hydrophilic micro-rough surface can significantly improve surface energy and capillary adsorption capacity, so that the bacterial liquid can be quickly, uniformly and stably attached to the protrusion and depression structure, further improving the consistency of dipping and the repeatability of transfer, and reducing the liquid volume fluctuation caused by surface tension differences.

[0047] A continuous, closed annular baffle 4 is provided around the outer periphery of the inoculation section. The annular baffle 4 surrounds the entire area of ​​the dot matrix protrusions 3, forming an outer protective structure. The height of the annular baffle 4 is lower than the height of the protrusions 31, creating a significant height difference between the bottom end face of the baffle and the recessed portion 32 at the top of the protrusion. This height difference effectively prevents the non-working edge of the inoculation block body 1 from accidentally contacting the porous liquid storage pad or the surface of the culture dish during movement, dipping, and imprinting, preventing edge contamination, cross-contamination, and large-area liquid spot contamination, while not affecting the normal contact, dipping, and imprinting actions of the protrusions.

[0048] This embodiment achieves stable bacterial liquid pick-up, regular dot matrix imprinting, smooth detachment of the inoculation block, and full-process anti-contamination protection through a combination of external size ratio constraints, through-type pressure relief structure, hydrophilic micro-rough capillary structure, and outer ring edge protection. It is particularly suitable for high-throughput, automated, and standardized bacterial dot matrix inoculation scenarios.

[0049] Example 2 Please refer to Figure 3 This embodiment provides an imprint-type bacterial liquid array inoculation device.

[0050] The device adopts a modular integrated design, with a compact structure, reasonable layout, and smooth operation. It is specially adapted to and uses the imprinted bacterial solution matrix inoculation block described in Example 1. It can realize the fully automated operation of the entire process from inoculation block supply, negative pressure pickup, bacterial solution supply, gradient dilution, capillary dipping, matrix imprinting, image detection, data traceability to the disposal and recycling of inoculation blocks after use, which greatly improves inoculation efficiency and experimental consistency.

[0051] The feeding mechanism in the device adopts a clip-type feeding unit, which is stable in structure, continuous in feeding, and precise in positioning. The clip-type feeding unit is closely arranged with the working position of the picking mechanism 5, which reduces the picking stroke and action time. It can continuously, orderly and uniformly output the inoculation block body 1, meet the needs of long-term high-throughput automated operation, and can also be compatible with various feeding forms such as drawer pallet type, stacked silo type, and turntable type according to the site conditions.

[0052] The pickup mechanism 5 is the core actuator of the device, equipped with a high-precision negative pressure nozzle, a vision inspection unit, a negative pressure detection unit, and an anti-inversion recognition unit. It has multiple functions, including pickup, positioning, posture verification, and moving imprinting. Driven by the control mechanism, the negative pressure nozzle can make stable three-axis movements between the supply station, the liquid supply station, and the culture dish carrying station. It operates smoothly, has accurate positioning, and high repeatability, and can reliably complete the entire process of pickup, dipping, imprinting, and placement.

[0053] The liquid supply mechanism is located directly below the moving path of the picking mechanism 5, which facilitates the negative pressure nozzle to quickly drive the inoculation block to the liquid supply position. In this embodiment, the liquid supply mechanism uses a porous liquid storage pad 6 as a carrier for bacterial liquid. The porous liquid storage pad 6 is uniformly wetted and carries bacterial liquid 7, which can provide a stable, uniform and quantitative supply of bacterial liquid to the inoculation block, avoiding problems such as excessive dipping, dripping and splashing caused by the free liquid surface.

[0054] The dilution mechanism is directly connected to the liquid supply mechanism, which can automatically complete the gradient dilution of bacterial solutions according to the test requirements. By precisely controlling the dilution ratio and mixing process, a series of bacterial solutions with different concentration gradients can be obtained quickly to meet various application scenarios such as drug sensitivity testing, inhibition zone testing, and growth curve determination.

[0055] The replenishment / pad replacement mechanism is arranged adjacent to the porous storage pad 6. It can automatically determine the remaining amount and uniformity of bacterial solution in the storage pad based on the preset number of dips, real-time weight signal change detection signal or image detection signal, and automatically trigger the replenishment or pad replacement action to always maintain a stable and consistent liquid supply state and avoid inoculation failure due to insufficient bacterial solution or uneven distribution.

[0056] The waste / recycling mechanism is located at the end of the movement path of the picking mechanism 5. It adopts a biohazardous waste bin structure and is specifically used to collect and temporarily store the inoculation block body 1 after use. This enables centralized harmless treatment of consumables after use, avoiding environmental pollution and cross-infection. It can also be switched to an offline sterilization and recycling bin as needed to achieve sterilization and reuse of the inoculation block.

[0057] The vision and traceability system uses a high-definition image acquisition and data processing unit to collect, detect, and analyze the dot matrix status after imprinting in real time, including the accuracy of dot matrix position, the integrity of arrangement, the uniformity of liquid spot diameter, and whether there are any abnormalities such as continuous patches, missing parts, or offsets. It also records the inoculation batch, bacterial concentration, imprint coordinates, liquid spot data, and petri dish number simultaneously, forming a complete and traceable experimental data chain that meets laboratory standards and data audit requirements.

[0058] The control mechanism is the core of the entire device, electrically connected to the pickup mechanism 5, liquid supply mechanism, dilution mechanism, liquid replenishment / pad replacement mechanism, waste / recycling mechanism, and vision and traceability mechanism. It coordinates the timing, speed, positioning, and triggering logic of each mechanism. The control mechanism precisely controls the imprinting pressure, imprinting position, holding time, and number of repeated imprints at the imprinting inoculation section, ensuring consistent parameters and stable results for each imprinting action.

[0059] In this embodiment, the porous reservoir pad is made of sterile PVA sponge, which has good hydrophilicity, uniform pores, and stable adsorption. The overall pore size is controlled at 100 μm, and the overall thickness is 3 mm. The pre-added bacterial solution volume is set to 100 μL, ensuring uniform distribution and stable release. The porous reservoir pad adopts a matrix-style partitioned structure, which can simultaneously hold multiple different samples or bacterial solutions of different concentrations, adapting to parallel dipping of multiple samples and high-throughput screening. The dipping contact pressure is set to 0.1 N, and the contact time is set to 0.5 s. These gentle and stable contact parameters ensure that the bacterial solution is fully, uniformly, and consistently dipped without damaging the bacteria or disrupting the reservoir pad structure.

[0060] This embodiment achieves fully automated, unmanned operation of the entire process from material supply, dipping, imprinting, testing to traceability and recycling through multi-mechanism collaborative control. It has significant advantages such as high inoculation throughput, good site consistency, low contamination risk, and full data traceability, and is widely applicable to scenarios such as colony counting, clone screening, drug sensitivity testing, microbial culture and condition optimization.

[0061] Example 3 Please refer to Figure 1 , Figure 2 , Figure 3 This embodiment provides a method for inoculating bacterial suspensions using an imprinting method.

[0062] This method is based on the aforementioned imprinted bacterial suspension array inoculation block and inoculation device. It features a standardized process, controllable parameters, and good repeatability, and can stably complete high-throughput, standardized, and pollution-free automated bacterial suspension array inoculation.

[0063] The usage process of the above embodiments is as follows: S1 supplies inoculation blocks. The operator pre-loads the sterilized inoculation block body 1 neatly into the magazine feeding unit of the supply mechanism. The inoculation blocks are arranged in an orderly manner with a uniform posture. The supply mechanism automatically transports the inoculation blocks to the picking position in sequence, completing the automated supply preparation and providing a stable source of consumables for continuous operation.

[0064] S2 prepares the liquid supply pad. The clean and sterile porous liquid storage pad 6 is placed stably in the designated bearing position inside the liquid supply mechanism. The liquid supply mechanism automatically positions and fixes the porous liquid storage pad 6. Then, 100μL of bacterial solution 7 is added into the porous liquid storage pad 6. Under capillary action, the bacterial solution quickly and evenly wets the entire pore of the porous liquid storage pad 6, so that the bacterial solution is distributed without discontinuity, enrichment, or drying areas, and maintains a stable liquid supply state.

[0065] S3 negative pressure pickup: Under the drive of the control mechanism, the pickup mechanism 5 moves the negative pressure nozzle to the pickup position of the supply mechanism. The negative pressure nozzle is pressed against the smooth pickup part on the upper surface of the inoculation block body 1, and the negative pressure is activated to form a stable adsorption. At the same time, the posture of the inoculation block is double-checked by the visual detection unit and the anti-inversion structure to ensure that the position is correct, there is no tilt, and there is no reverse direction, thus completing reliable pickup.

[0066] S4 capillary dipping: The picking mechanism 5 drives the picked-up inoculation block body 1 to move smoothly above the liquid supply mechanism and slowly descends along a preset path, so that the dot matrix protrusions 3 on the lower surface of the inoculation block contact the surface of the porous liquid storage pad 6 with a constant pressure of 0.1N and maintain the contact time for 0.5s. During this process, the recessed part 32 at the top of the protrusion stably and quantitatively picks up the bacterial liquid 7 under the combined action of the hydrophilic micro-rough surface and capillary action, ensuring that the amount picked up by each protrusion is highly consistent.

[0067] After the S5 mobile positioning is completed, the picking mechanism 5 drives the inoculation block body 1 to rise smoothly and move quickly to the position directly above the culture dish. The vision and position control system accurately positions the inoculation block and the inoculation area of ​​the culture dish, ensuring that the imprinting dot matrix position is accurate and without deviation.

[0068] S6 dot matrix imprinting: The control mechanism drives the pickup mechanism 5 and the inoculation block body 1 to press downwards at a uniform speed, so that the dot matrix protrusions 3 make uniform contact with the surface of the culture dish. The bacterial solution 7 carried on the dot matrix protrusions 3 is smoothly transferred to the surface of the culture dish under pressure, forming a regular and neat colony matrix. At the same time, the pressure relief groove 2 on the lower surface of the inoculation block body is connected to the outside atmosphere through the through opening, quickly breaking the negative pressure adsorption between the inoculation block and the culture medium, preparing for smooth detachment.

[0069] After the S7 static absorption and imprinting action is completed, the inoculation block body 1 is kept in a static state under the control of the control mechanism and is not lifted immediately, so that the bacterial solution transferred to the surface of the culture dish can fully penetrate into the agar, ensuring complete absorption of the bacterial solution, fixed position, no diffusion, no shrinkage, and improving the consistency of colony morphology.

[0070] After the S8 image traceability is completed, the vision and traceability mechanism immediately acquires and intelligently analyzes the imprinted dot matrix, detects the integrity of the dot matrix arrangement, the uniformity of the liquid spot diameter, the accuracy of the dot position, and whether there are any abnormalities such as continuous patches or missing parts. At the same time, the batch, concentration, location, diameter, culture dish number and other information are stored in the system to form a complete traceable record.

[0071] After image detection is completed, the picking mechanism 5 moves the used inoculation block body 1 above the waste / recycling mechanism, closes the negative pressure to release the inoculation block, and causes it to fall into the biohazardous waste bin or offline sterilization and recycling bin, completing a single inoculation process. Subsequently, the picking mechanism 5 automatically returns to the supply mechanism to pick up the next inoculation block body 1, entering the next round of inoculation cycle, realizing continuous and uninterrupted automated operation.

[0072] The entire inoculation process in this embodiment is fully automated and controlled by the device, requiring no manual intervention. The actions are standardized, the parameters are constant, and the process is standardized, significantly reducing human error and the risk of contamination. The inoculation efficiency is high, the results are consistent, and the data is reliable. It can reliably meet the high-throughput, standardized, and traceable inoculation needs of modern microbiology laboratories.

[0073] Obviously, the above embodiments of the present invention are merely examples for clearly illustrating the technical solutions of the present invention, and are not intended to limit the specific implementation of the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the claims of the present invention should be included within the protection scope of the claims of the present invention.

Claims

1. An embossed bacterial liquid dot array inoculation block, characterized in that, The inoculation block body (1) is a thin sheet-like structure, and the inoculation block body (1) is divided into the following parts: A smooth pickup section is located on the upper surface of the inoculation block body (1) and has a smooth pickup area; The inoculation section, located on the lower surface of the inoculation block body (1), includes at least one pressure relief groove (2) and dot matrix protrusions (3) located on both sides of the pressure relief groove (2), wherein: The pressure relief channel (2) extends from the middle of the lower surface to both sides of the periphery of the lower surface to form a through opening; The dot matrix protrusions (3) include several protrusions (31). The protrusions (31) are evenly distributed in a symmetrical matrix on both sides of the pressure relief channel (2). Each protrusion (31) has a concentric recess (32) at the top center.

2. The imprinted bacterial suspension array inoculation block as described in claim 1, characterized in that: The inoculation block body (1) is a thin sheet structure adapted to the shape of the culture dish; the depth of the pressure relief groove (2) is less than the height of the dot matrix protrusion (3).

3. The imprinted bacterial suspension array inoculation block as described in claim 1, characterized in that: The pickup area is a flat surface, a shallow concave surface, or a mating surface that fits a sealing ring, so as to achieve negative pressure mating with the nozzle; The pickup area is equipped with a QR code, color mark, batch identification code, asymmetric error-proof notch or single-sided chamfer for positioning, identification or error prevention.

4. The imprinted bacterial suspension array inoculation block as described in claim 1, characterized in that: The pressure relief channel (2) is one or more of the following: straight channel, broken channel, curved channel, cross channel or radial channel; The through opening is a straight opening, an arc opening or a geometrically regular opening, and the through opening extends through to the side wall of the inoculation block body (1), so that the pressure relief groove (2) forms an air channel communicating with the outside. The pressure relief channel (2) is located between the dot matrix protrusions and avoids the inoculation position of the dot matrix protrusions. Its depth is greater than the depth of the recess (32) so as to ensure that when the inoculation block body (1) is lifted, external air enters between the lower surface of the inoculation block body and the culture medium through the through opening to break the negative pressure.

5. The imprinted bacterial suspension array inoculation block as described in claim 1, characterized in that: The protrusion (31) is a cylindrical, frustum-shaped, or hemispherical structure; The recess (32) is one or more of the following: micro-blind hole, micro-through hole, annular micro-groove, cross micro-groove or radial micro-groove.

6. The imprinted bacterial suspension array inoculation block as described in claim 5, characterized in that: The dot matrix protrusion (3) is also provided with a hydrophilic micro-rough surface, and the hydrophilic micro-rough end face is formed on the end face of the protrusion (31) and the inner wall of the recess (32); The hydrophilic micro-roughened end face is one or more of the following: a frosted surface, a sandblasted surface, a laser micro-etched surface, a chemical micro-etched surface, a plasma-treated surface, or a UV ozone-treated surface.

7. The imprinted bacterial suspension array inoculation block as described in claim 1, characterized in that: An annular baffle (4) is provided on the outer periphery of the imprinting inoculation part. The annular baffle (4) is arranged around the dot matrix protrusion (3). The height of the annular baffle (4) is lower than the height of the protrusion (31). A height difference is formed between the bottom end face of the annular baffle (4) and the recess (32) of the protrusion (31) to prevent the non-working surface of the inoculation block body (1) from accidentally contacting the porous liquid storage pad (6) and without affecting the normal imprinting inoculation of the protrusion (31).

8. An embossing type bacterial liquid dot array inoculation device, characterized by, The device employs the imprinted bacterial suspension array inoculation block as described in any one of claims 1-7, characterized in that: the device comprises: The supply mechanism is used to supply the inoculation block body (1), including a clip type, drawer tray type, stacked hopper type or turntable type supply unit, and the supply unit is arranged adjacent to the working position of the picking mechanism (5). Pick-up mechanism (5) is used to pick up the smooth pickup part by negative pressure, including negative pressure nozzle, vision / negative pressure / height detection unit and anti-inversion recognition unit. The negative pressure nozzle moves above the supply mechanism, liquid supply mechanism and culture dish support module. The liquid supply mechanism is a porous liquid storage pad (6), and each porous liquid storage pad (6) is filled with bacterial liquid (7). The liquid supply mechanism is located below the moving path of the picking mechanism (5). A dilution mechanism is used to perform gradient dilution of the bacterial solution (7) from the liquid supply mechanism, and is connected to the liquid supply mechanism. The liquid replenishment / pad replacement mechanism is used to replenish / replace the liquid supply mechanism. It is set up adjacent to the liquid supply mechanism and is triggered by the number of dips, weight signal changes, or image signals to replenish / replace the liquid. The disposal / recycling mechanism is used to dispose of / recycle the inoculation block body (1) after inoculation, including a biohazardous waste bin / offline sterilization and recycling bin, which is located at the end of the movement path of the picking mechanism (5); A visual and traceability system is used to record relevant information about the inoculation process, including batch number, concentration, imprint location, spot diameter, and petri dish number. The control mechanism is electrically connected to the pickup mechanism (5), the liquid supply mechanism, the dilution mechanism, the liquid replenishment / pad replacement mechanism, the waste / recycling mechanism, and the vision and traceability mechanism, respectively, and is used to control the imprinting pressure, position, dwell time and number of repetitions of the imprinting inoculation part, and coordinate the timing of actions.

9. The imprint bacterial liquid dot array inoculation device of claim 8, wherein : The porous liquid storage pad (6) in the liquid supply mechanism is one or more of the following: sterile sponge, PVA sponge, non-woven fabric, cellulose pad, microporous polyurethane pad, microporous silicone pad or sintered porous plastic pad. Alternatively, the porous liquid storage pad (6) may be configured as a strip, matrix, ring, or multi-compartment tray partition structure.

10. A method for imprinting bacterial liquid dot array inoculation, using the imprinting bacterial liquid dot array inoculation device according to any one of claims 8-9, characterized in that: Includes the following steps: S1. Supplying inoculation blocks: The sterilized inoculation block body (1) is loaded into the supply mechanism and supplied in the form of a clip, drawer tray, stacked hopper or turntable. S2. Preparation of the liquid supply pad: Place the porous liquid storage pad (6) into the liquid supply mechanism and add a predetermined volume of bacterial solution (7) into the porous liquid storage pad (6); S3, Negative pressure pickup: The inoculation block body (1) is picked up from the supply mechanism through the negative pressure nozzle of the pickup mechanism (5), and the correct posture is confirmed by vision, negative pressure or foolproof structure. S4, capillary dipping: The picking mechanism (5) drives the inoculation block body (1) to descend, so that the dot matrix protrusions (3) of the imprinted inoculation part contact the porous liquid storage pad (6) with a predetermined pressure, and pick up the bacterial liquid (7) through capillary action. S5, Moving and Positioning: The picking mechanism (5) moves the inoculation block body (1) to the culture dish; S6, Dot Imprinting: The control mechanism drives the inoculation block body (1) to press down, so that the dot protrusion (3) contacts the culture dish with a predetermined pressure and residence time, and the bacterial solution (7) is transferred to form a regular colony dot array; at the same time, the pressure relief channel (2) maintains air communication through the through opening, and breaks the negative pressure between the bottom surface and the culture dish. S7. Static absorption: After imprinting, allow the inoculum block body (1) to stand for a short time, allowing the bacterial solution (7) to be fully absorbed by the culture medium. S8. Image traceability: Detects the position, integrity, liquid spot diameter, and presence of continuous liquid droplets of the imprinted dot matrix through vision and traceability mechanisms; S9. Discard / Recycle: The picking mechanism (5) releases the used inoculation block body (1) to the discard / recycle mechanism, and then picks up the next inoculation block body (1) to enter the next cycle.

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