Automatic inoculation method and inoculation device for pouring method in microbe counting

Abstract

The invention relates to an automatic inoculation method and device for a pouring method in microbiological counting, and belongs to the field of microbiological devices.The automatic inoculation method comprises the following steps that a plurality of culture dishes added with sample liquid are placed in a lower dish channel and transferred to a dish receiving station; in the dish receiving station, the bottoms of the culture dishes are moved downwards to a filling disc through a guide cylinder material plate I, and the filling disc transfers the culture dishes to the inoculation station; in the inoculation station, a culture medium is injected into the culture dish bottom, after a filling disc swings and shakes clockwise and anticlockwise in a reciprocating mode, a control wheel drives the culture dish bottom to rotate, and the culture dish bottom enters a transition station after rotating; the transition station is used for transferring the culture dishes into the code printing station; the bottom of the culture dish is coded and transferred to the stacking station; after the bottoms of the culture dishes are supported and buckled with the culture dish covers, the culture dishes are dragged into the stacking channels, and through the steps, dish receiving, inoculation, transition, code printing and stacking of the culture dishes in the multiple dish discharging channels are completed. Human factor interference in the inoculation process is reduced to the greatest extent, and the inoculation process of a pouring method can be efficiently completed.

Description

Technical Field

[0001] The invention relates to an automatic inoculation method and an inoculation device of a pouring method in microbial counting, belonging to the field of microbiological devices. Background Art

[0002] In the actual inspection and testing work of the existing technology, personnel prepare the sample liquid, drip it into the culture dish, and make gradient marks one by one. Pour the sterilized agar culture medium into the culture dish, and manually shake the culture dish until the sample liquid and the culture medium in the culture dish are evenly mixed. During this period, the culture medium should be maintained in the range of 45-50°C. If the temperature of the culture medium is too low, the agar will clot and it will be unfavorable for the growth of the colony. If it is too high, it will scald and burn the bacteria. However, depending on the different samples, the time and degree of manual shaking of the culture dish to achieve uniform mixing are difficult to control and unify standards. It is time-consuming and labor-intensive, with large batch errors, no traceability, and a high chance of human cross-contamination. A small number of equipment rely on imported equipment, and the equipment functions fail to take into account the requirements of the inspection items of the "Chinese Pharmacopoeia", and are expensive and have high maintenance costs.

[0003] Therefore, the current market lacks an automated inoculation method and inoculation device for microbial counting using the pouring method that can achieve high automation, high precision and safety. This technological gap not only affects the work efficiency of the laboratory, but also restricts the further development of the biomedicine field to a certain extent. Summary of the Invention

[0004] Based on the existing problems in the prior art, the sample liquid is easily affected by human factors such as the operating level and proficiency of the production personnel during the inoculation process, resulting in uneven mixing of the sample liquid and the culture medium, easy coagulation of the culture medium, low inoculation efficiency, unstable inoculation quality and other problems. The present invention provides an automated inoculation method and inoculation device for the pouring method in microbial counting, which minimizes the interference of human factors in the inoculation process, can efficiently complete the inoculation process of the pouring method, and achieves uniform mixing of the sample liquid and the culture medium.

[0005] The technical solution adopted by the present invention is: an automated inoculation method of the pouring method in microbial counting, comprising the following steps: Step 1, setting a V-shaped culture dish channel in a silo as a stacking channel, and setting the remaining multiple V-shaped culture dish channels as lower dish channels, and respectively loading multiple culture dishes with sample liquid into the lower dish channels; starting the machine, with the first lower dish channel corresponding to the culture dish inlet hole, the stacking channel corresponding to the culture dish outlet hole, and two gripper arms gripping the first culture dish in the first lower dish channel; Step 2: At the dish receiving station, the receiving motor assembly is started, the guide plate I moves upward and extends out of the culture dish hole in area A of the filling tray, and is placed under the culture dish inlet hole of the silo support plate. The hand claw stepper motor is started, and the hand claw cam drives the two hand claws to expand. The first culture dish falls on the guide plate I. The hand claw cam drives the two hand claws to retract and clamp the second culture dish. The guide plate I moves downward and moves the first culture dish down to the culture dish hole in area A of the filling tray. The culture dish cover is placed on the raised platform of the canning tray, and the culture dish bottom falls on the table main board in the culture dish half-opening hole II 5-2-1; Step 3: At the inoculation station, the filling tray motor assembly starts and the filling tray rotates counterclockwise. When the bottom of the first culture dish enters the inoculation station, the filling tray stops rotating and the filling peristaltic pump assembly starts. The culture medium, which is kept at a constant temperature by the heating thermostat, is injected into the bottom of the culture dish through the filling head. The filling tray swings back and forth clockwise and counterclockwise. After the back and forth swing, the O-ring of the control wheel drives the bottom of the culture dish to rotate, and the rotation stops. At the same time, the second culture dish is transferred to the dish receiving station, and the second culture dish is moved down to the culture dish hole in area B of the filling tray. Repeat step 2 to complete the dish receiving work, and the two gripper arms grip the third culture dish; Step 4: At the transition station, the filling tray motor assembly starts, the filling tray rotates counterclockwise, the first culture dish is transferred to the transition station, the second culture dish is transferred to the inoculation station, the filling tray stops rotating, and step 3 is repeated. After the culture medium is injected into the second culture dish, the filling tray drives the bottom of the second culture dish and the bottom of the first culture dish to rock back and forth clockwise and counterclockwise at the same time. The rocking stops, and the O-ring of the control wheel drives the bottom of the second culture dish to rotate, and the rotation stops; at the same time, the third culture dish is transferred to the receiving dish station, and the third culture dish is moved down to the culture dish hole in area C of the filling tray. Step 2 is repeated to complete the receiving dish work, and the two gripper arms grip the fourth culture dish; Step 5: At the coding station, the filling tray motor assembly is started, the filling tray rotates counterclockwise, the bottom of the first culture dish is transferred to the printing station, the bottom of the second culture dish is transferred to the transition station, and the bottom of the third culture dish is transferred to the inoculation station. The filling tray stops rotating, and step 3 is repeated. After the injection of the culture medium of the third culture dish is completed, the filling tray drives the bottoms of the third, second, and first culture dishes to swing back and forth clockwise and counterclockwise at the same time. The reciprocating swinging stops, and the O-ring of the control wheel drives the bottom of the third culture dish to rotate. The rotation stops, and the coding of the bottom of the first culture dish is completed; at the same time, the fourth culture dish is transferred to the receiving dish station, and the fourth culture dish is moved down to the culture dish hole in area D of the filling tray. Step 2 is repeated to complete the receiving dish work, and the two gripper arms grip the fifth culture dish; Step 6: Stacking station, the filling tray motor assembly is started, the filling tray rotates counterclockwise, the bottom of the first culture dish is transferred to the stacking station, the bottom of the second culture dish is transferred to the coding station, the bottom of the third culture dish is transferred to the transition station, and the fourth culture dish is transferred to the inoculation station. The filling tray stops rotating, and the guide plate II moves upward to lift the bottom of the first culture dish in the culture dish hole and buckle it with the culture dish cover. Then, it is passed through the two U-shaped culture dish supporting plates in the culture dish outlet hole of the silo supporting plate and supported into the stacking channel of the silo. The front ends of the two U-shaped culture dish supporting plates 4-4 support the culture dishes in the stacking channel. Repeat step 3 to complete the injection of culture medium into the fourth culture dish. The filling tray drives the bottoms of the fourth, third, and second culture dishes to swing back and forth clockwise and counterclockwise simultaneously. The swinging stops, and the O-ring of the control wheel drives the bottom of the fourth culture dish to rotate, and the rotation stops. The bottom of the second culture dish is coded. At the same time, the fifth culture dish enters the receiving station and moves down to the culture dish hole in area E of the filling tray. Repeat step 2 to complete the receiving work, and the two gripper arms grip the sixth culture dish. Step 7: Repeat steps 2 to 6 to sequentially complete the stacking of the second culture dish bottom, the coding and stacking of the third culture dish bottom, the transition, coding, and stacking of the fourth culture dish, the inoculation, transition, coding, and stacking of the fifth culture dish bottom, and the connection, inoculation, transition, coding, and stacking of the sixth to multiple culture dishes. Step 8: After the multiple culture dishes in the first dish lowering channel of the hopper have finished moving downward, the hopper rotates counterclockwise, and the first dish lowering channel becomes a stacking channel; Through steps 2 to 6, the receiving, inoculation, transfer, coding and stacking of multiple culture dishes in the next dish channel are completed in sequence.

[0006] The clockwise and counterclockwise reciprocating rocking of the filling plate described in step three includes a rocking angle of 1-90 degrees, a rocking speed of 10-500 rpm, a rocking interval of 0.1-3 seconds, and a rocking frequency of 1-20 times; the control wheel drives the bottom of the culture dish to rotate in the filling plate at an angle of 60-90 degrees; and the constant temperature of the culture medium is 36°C-50°C.

[0007] An inoculation device, a constant temperature filling device, a vertical plate with an ultraviolet lamp, a safety cover and a U-shaped enclosure are respectively fixed on the main surface of the shell table, and a silo supporting plate is fixed on the vertical plate and the U-shaped enclosure, and a silo motor assembly and two hand claw arms are fixed under the silo supporting plate, and the shaft of the silo motor assembly passes through the silo supporting plate and is fixed to the silo arranged on the silo supporting plate, so as to drive the silo to rotate; a filling disk motor assembly, a hand claw motor assembly, a material receiving motor assembly, a material ejecting motor assembly, a culture medium uniformity control device and an inkjet printer are respectively fixed under the main surface of the table, and the shaft of the filling disk motor assembly passes through the main surface of the table and is fixed to the filling disk arranged on the main surface of the table, so as to drive the filling disk to rotate; the hand claw cam of the hand claw motor assembly extends out of the main surface of the table and is slidably connected with the two hand claw arms, so as to drive the two hand claw arms to expand outward or retract inward, and the guide cylinder material plate I of the material receiving motor assembly and the guide cylinder material plate of the material ejecting motor assembly Ⅱ The control wheels of the culture medium uniformity control device extend out of the main board surface of the table respectively. The control wheels are set in the opening slot of the constant temperature filling device. The control wheels and the filling head guide plate I of the constant temperature filling device correspond to the culture dish inlet hole of the silo support plate. Ⅱ Corresponding to the outlet hole of the culture dish, the inkjet printer head of the inkjet printer is arranged in the inkjet window; the filling peristaltic pump assembly is fixed on the inner bottom plate of the shell, the peristaltic pump head of the filling peristaltic pump assembly is exposed outside the shell panel, and the discharge pipe mouth on the peristaltic pump head is connected with the filling head of the constant temperature filling device; the operation screen, USB data interface and external peristaltic pump interface of the PLC all-in-one are respectively arranged on the shell panel; the PLC all-in-one is respectively connected with the filling peristaltic pump assembly, constant temperature filling device, silo motor assembly, filling tray motor assembly, gripper motor assembly, material receiving motor assembly, top material motor assembly, culture medium uniformity control device, ultraviolet lamp, operation screen, inkjet printer, USB data interface and external peristaltic pump interface.

[0008] The technical effect produced by the present invention is: the constant temperature filling device is provided with a filling head bracket, and a polyimide heating plate and a thermocouple are installed at the lower end of the filling head bracket. The surface temperature of the filling head bracket is adjusted by controlling the start and stop of the heating plate. The filling head for conveying the culture medium is a thin-walled stainless steel filling head, and its outer wall fits the surface of the filling head bracket to transfer the temperature to the culture medium, so that the culture medium maintains a constant temperature during the filling process. The filling temperature can be set by the user within the range of 35-50° to keep the culture medium at a suitable temperature.

[0009] After the culture medium is added, the filling plate drives the culture dish to swing back and forth in the clockwise and counterclockwise directions. The first time the filling plate swings back and forth 1-90 degrees clockwise and counterclockwise to make the sample liquid in the bottom of the culture dish diffuse laterally in the culture medium. After the filling plate stops swinging, the pulse control of the stepper motor and the friction between the O-ring on the control wheel and the contact part of the culture dish bottom are controlled to drive the bottom of the culture dish to rotate 60-90 degrees in the filling plate. The second and third time the filling plate swings back and forth 1-90 degrees clockwise and counterclockwise to make the sample liquid in the bottom of multiple culture dishes diffuse laterally in the culture medium again. Since the bottom of the culture dish rotates 60-90 degrees in the filling plate, the filling plate swings back and forth multiple times. swing The shaking makes the sample liquid diffuse vertically. The sample liquid in the bottom of the culture dish is subjected to centrifugal action and is fully dispersed to the edge of the culture dish, so that the sample liquid and culture medium are evenly mixed, providing single colony conditions for colony counting and separation after cultivation.

[0010] After the sample liquid and culture medium are fully mixed, the filling tray rotates to the coding position, and the system automatically assigns a serial number mark to the culture dish. After marking, the filling tray rotates to the cover and stacking position for arrangement, and the operator takes it out for subsequent culture.

[0011] The silo and filling tray are respectively linked by a double-outlet turntable motor, which overcomes the shortcomings of the existing technology that the silo and filling tray are respectively coordinated with the motor gear through the silo and filling tray gears, resulting in errors in the rotation of the silo and filling tray, and poor synchronization between the culture medium out of the silo and the filling tray receiving the culture medium. It can accurately achieve the consistency of the synchronization between the culture medium out of the silo and the filling tray receiving the culture medium.

[0012] The structural design of the gripper cam and gripper arm enables precise control of the discharge of each culture dish from the lower dish channel of the silo. The structural design of the rotating silo combined with the rotating filling plate saves space while ensuring that the opening and closing of the culture dish lids are completed in the smallest range, effectively reducing the risk of contamination.

[0013] When the culture medium is inoculated, the safety cover and the U-shaped enclosure form a sealed inoculation area. The UV lamp is linked to the filling process to ensure that the UV lamp is always on during the entire inoculation process, thereby ensuring the sterile operation of the inoculation area and protecting the operator from liquid splashes and ultraviolet rays.

[0014] The inoculation device has high inoculation efficiency and can complete more than 500 inoculations per hour. The efficiency is more than three times higher than manual mixing, effectively reducing the labor intensity of experimenters. The inoculation process is standardized, which is superior to manual operation and realizes the traceability of the experimental process. The entire inoculation process is completed in a relatively closed sterile space without human intervention, effectively reducing the risk of cross-contamination and meeting the requirements of the GMP quality management system.

[0015] The device automatically completes a series of tasks including receiving plates, opening lids, filling culture medium, mixing sample liquid and culture medium, coding and marking, closing lids, and stacking and arranging culture medium plates, complying with the relevant requirements of the plate pouring method in national standards such as GB / T 4789 and the Chinese Pharmacopoeia. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 A schematic diagram of the overall structure of the inoculation device of the present invention; Figure 2 for Figure 1 Schematic diagram of the internal structure; Figure 3 for Figure 2 The structure is shown in side view; Figure 4 It is a structural schematic diagram of the silo of the present invention; Figure 5 This is a schematic structural diagram of the turntable on the silo of the present invention; Figure 6 This is a schematic structural diagram of the lower turntable of the silo of the present invention; Figure 7 It is a structural schematic diagram of the filling tray of the present invention; Figure 8 This is a schematic structural diagram of the silo support plate of the present invention; Figure 9 for Figure 8 Bottom view of the structure; Figure 10 This is a schematic diagram of the specific positions of the gripper in-position sensor and the gripper cam of the present invention; Figure 11 This is a schematic diagram of the specific position of the coding window of the present invention; Figure 12 This is a structural diagram of the silo support plate and the table main board being fixed to each other in the present invention; Figure 13 This is a schematic structural diagram of the filling peristaltic pump assembly of the present invention; Figure 14 It is a structural schematic diagram of the constant temperature filling device of the present invention; Figure 15 for Figure 14 Bottom view of the assembled constant temperature filling device; Figure 16 This is a schematic structural diagram of the silo motor assembly of the present invention; Figure 17 This is a schematic structural diagram of the filling tray motor assembly of the present invention; Figure 18 This is a schematic structural diagram of the gripper motor assembly of the present invention; Figure 19 A top view of the gripper motor assembly of the present invention; Figure 20This is a structural diagram of the material receiving motor assembly of the present invention; Figure 21 for Figure 20 Schematic diagram of the structural breakdown of the material receiving motor assembly; Figure 22 This is a schematic structural diagram of the ejector motor assembly of the present invention; Figure 23 Schematic diagram of the structure of the culture medium uniformity control device of the present invention; Figure 24 for Figure 23 Schematic diagram of the structure breakdown of the culture medium uniformity control device; Figure 25 A circuit connection diagram for realizing the vaccination device of the present invention; Figure 26 This is a schematic structural diagram of a culture dish according to an embodiment of the present invention; Figure 27 A screenshot showing the comparison of the mixing degree of the culture medium and the straw mushroom dark soy sauce stock solution by shaking 10 times in the first lower plate channel during the inoculation process using Example 1 of the inoculation device and manually pouring the culture medium into the plate and adding a 10-fold dilution; Figure 28 A screenshot showing the comparison of the mixing degree of the culture medium and the straw mushroom dark soy sauce stock solution by shaking 15 times in the second lower plate channel during the inoculation process of Example 1 of the inoculation device and manually pouring the culture medium into the plate and adding a 10-fold dilution; Figure 29 A screenshot showing the comparison of the mixing degree of the culture medium and the straw mushroom dark soy sauce stock solution by shaking 20 times in the third lower plate channel during the inoculation process of Example 1 of the inoculation device and manually pouring the culture medium into the plate and adding a 10-fold dilution; Figure 30 A screenshot showing the comparison of the mixing degree of the culture medium and flour by shaking 10 times in the first lower plate channel during the inoculation process of Example 2 of the inoculation device and manually pouring the culture medium and flour into the plate with a 10-fold dilution; Figure 31 A screenshot showing the comparison of the mixing degree of the culture medium and flour by shaking 15 times in the second lower plate channel during the inoculation process using Example 2 of the inoculation device and manually pouring the culture medium and flour into the plate with a 10-fold dilution; Figure 32 This is a screenshot of the comparison results of the mixing degree of the culture medium and flour by shaking 20 times in the third lower dish channel during the inoculation process of Example 2 of the inoculation device and manually pouring the culture medium and flour into the plate with a 10-fold dilution. DETAILED DESCRIPTION

[0017] like Figures 1 to 25As shown, an inoculation device includes a shell 1, a silo 2, a silo guard 3, a silo support plate 4, a filling tray 5, a filling peristaltic pump assembly 6, a constant temperature filling device 7, a silo motor assembly 8, a filling tray motor assembly 9, a gripper motor assembly 10, a material receiving motor assembly 11, a material pushing motor assembly 12, a culture medium uniformity control device 13, a gripper arm 14, a vertical plate 15, a laser sensor 16, an operation screen 17, a safety cover 18, an inkjet printer 19, a U-shaped enclosure 20, a PLC all-in-one machine, a USB data interface and an external peristaltic pump interface.

[0018] The constant temperature filling device 7, the vertical plate 15, the two laser sensors 16, the bracket 10-5 with the gripper in-position sensor 10-4 and the U-shaped enclosure 20 are fixed to the main board 1-2 of the shell 1 by bolts respectively, the silo supporting plate 4 is fixed to the vertical plate 15, the four columns and the U-shaped enclosure 20, and the ultraviolet lamp 15-1 is fixed to the vertical plate 15; the gripper arm limiting plate 14-4 is fixed by bolts under the silo supporting plate 4, and two gripper arms 14 are provided between the gripper arm limiting plate 14-4 and the silo supporting plate 4, one end of the gripper arm 14 is provided with a bearing 14-3, and the other end is a gripper 14-1, and the middle of the two gripper arms 14 are connected to the silo supporting plate 4 by finger shafts 14-2 respectively. On the top, one end of the two gripper arms 14 is respectively provided with a bearing 14-3, and the grippers 14-1 at the other ends of the two gripper arms 14 are symmetrically arranged on both sides of the culture dish inlet hole 4-1 of the silo supporting plate 4, and the two gripper arms 14 are fixed under the silo supporting plate 4; the silo motor assembly 8 motor fixing sleeve Ⅰ8-3 is fixed to the underside of the silo supporting plate 4 by bolts, and the silo motor assembly 8 shaft passes through the center hole of the silo supporting plate 4 and is fixed to the shaft hole 2-1-1 of the lower turntable of the silo 2 set on the silo supporting plate 4. The silo motor assembly 8 rotates clockwise or counterclockwise to drive the silo 2 to rotate accordingly; the filling plate motor assembly 9 motor fixing sleeve Ⅱ9-3, the gripper motor assembly 10 and the gripper stepper motor 10-1 are fixed together. Lan, the material receiving motor assembly 11 screw motor bracket Ⅰ11-2, the top material motor assembly 12 screw motor bracket Ⅱ12-2, the culture medium uniformity control device 13 control motor board 13-4 and the bracket equipped with the inkjet printer 19 are respectively fixed to the bottom of the table main board 1-2 by bolts, and the filling disk motor assembly 9 shaft passes through the table main board 1-2 and is fixed to the filling disk 5 through hole 5-4 set on the table main board 1-2. The filling disk motor assembly 9 rotates clockwise or counterclockwise to drive the filling disk 5 to rotate accordingly; the hand claw motor assembly 10 hand claw cam 10-2 extends out of the table main board 1-2, and the two arc-shaped strip holes 10-2-1 on the hand claw cam 10-2 are respectively set on the bearings 1 of the two hand claw arms 14 4-3, the gripper in place baffle 10-3 corresponds to the gripper in place sensor 10-4 fixed on the bracket 10-5, the gripper motor assembly 10 and the gripper stepper motor 10-1 rotate clockwise or counterclockwise, and the two arc-shaped strip holes 10-2-1 of the gripper cam 10-2 cooperate with the bearings 14-3 of the two gripper arms 14, driving the two gripper arms 14 to expand outward or retract inward; the inkjet printer head of the inkjet printer 19 is set in the inkjet printer window 1-2-1; the multiple culture dish inlets 2-2-1 of the silo 2 are respectively concentric with the culture dish inlet hole 4-1, the culture dish outlet hole 4-2 of the silo support plate 4, the multiple culture dish holes 5-1-1 of the filling tray 5 and the multiple culture dish semi-opening holes II 5-2-1;The laser sensor 16 on the upper part of the bracket 10-5 is set corresponding to the raised platform 5-1-11 of the canning tray 5, and the laser sensor 16 on the lower part of the bracket 10-5 is set corresponding to the multiple semi-open holes Ⅱ5-2-1 of the culture dishes. The two laser sensors 16 are used to check whether the culture dish cover and the culture dish bottom in the filling tray 5 exist. If it is detected that there is no culture dish cover 21-1 or culture dish bottom 21-2 in the filling tray 5, the laser sensor 16 sends a signal to the PLC all-in-one machine, and the PLC all-in-one machine controls the culture medium inoculation device to stop working and eliminate the fault; the guide cylinder material plate Ⅰ11-5 of the material receiving motor assembly 11 extends upward from the table main board 1-2 and corresponds to the culture dish inlet hole 4-1 of the silo support plate 4, and the guide cylinder material plate Ⅱ12-5 of the top material motor assembly 12 extends upward from the table main board 1-2 and corresponds to the culture dish outlet hole 4-2 of the silo support plate 4; the control wheel 13-2 of the culture medium uniformity control device 13 extends upward from the table main board 1-2 is arranged in the opening slot 7-1-1 of the constant temperature filling device 7, the control wheel 13-2 and the filling head 7-2 of the constant temperature filling device 7 correspond to the filling tray 5 respectively, the filling peristaltic pump assembly 6 is fixed to the bottom plate 1-1 inside the shell 1 by bolts, the peristaltic pump head 6-2-1 of the filling peristaltic pump assembly 6 is exposed outside the panel 1-3 of the shell 1, the discharge pipe on the peristaltic pump head 6-2-1 is connected to the filling head 7-2 of the constant temperature filling device 7, the PLC all-in-one machine The operation screen 17, USB data port, and external peristaltic pump port are located on panel 1-3 of housing 1. The silo shield 3 is bolted to silo 2. The safety cover 18 is hinged to the main panel 1-2. The safety cover 18 snaps downward onto the main panel 1-2 and docks with the U-shaped panel 20, forming the culture medium filling area and the ultraviolet sterilization zone for the UV lamp 15-1. The safety cover 18 can be folded upward 90 degrees for troubleshooting.

[0019] The circuit connection is that the PLC all-in-one is respectively connected to the filling peristaltic pump assembly 6, the constant temperature filling device 7, the silo motor assembly 8, the filling tray motor assembly 9, the gripper motor assembly 10, the material receiving motor assembly 11, the material pushing motor assembly 12, the culture medium uniformity control device 13, the ultraviolet lamp 15-1, two laser sensors 16, the operation screen 17, the inkjet printer 19, the USB data interface and the external peristaltic pump interface.

[0020] The operation panel 17 is used to operate the culture medium inoculation device; the inkjet printer 19 is used to print online production information; a USB data port is used to export real-time filling data; and an external peristaltic pump port allows for simultaneous filling. The housing 1, silo 2, silo shield 3, silo support plate 4, and filling tray 5 are all constructed from high-strength aluminum alloy and SUS304 stainless steel, with a hard-anodized surface treatment for corrosion resistance, ease of cleaning, stability, and durability, meeting GMP requirements.

[0021] like Figures 4 to 6As shown, the silo 2 includes a silo lower turntable 2-1, a silo upper turntable 2-2, a silo support rod 2-3, a silo gear rod 2-4, a silo guide column 2-5, and a tension spring I 2-6; the silo upper turntable 2-2 is an annular disk, and a plurality of culture dish inlets 2-2-1 and a plurality of support rod holes I 2-2-3 are spaced apart along the annular surface of the annular disk, and each culture dish inlet 2-2-1 is provided with two V-shaped strip holes 2-2-2 on both sides; the silo lower turntable 2-1 is a circular disk, and an axis hole 2-1-1, a plurality of culture dish half-open outlets 2-1-2, a plurality of support rod holes II 2-1-3, a plurality of silo gear rod holes 2-1-4 and a plurality of silo guide column holes 2-1-5 are provided on the circular disk, and a plurality of silo guide columns 2-5 are respectively fixed on the plurality of silo guide column holes 2-1-5, and the lower ends of the plurality of silo support rods 2-3 are respectively fixed on the plurality of support rod holes II 2-1- 3, the upper ends of multiple silo support rods 2-3 are respectively fixed in multiple support rod holes I2-2-3, the lower ends of multiple silo gear rods 2-4 are respectively fixed in multiple silo gear rod holes 2-1-4, the upper ends of multiple silo gear rods 2-4 respectively extend out of multiple V-shaped bar holes 2-2-2, and the upper ends of the silo gear rods 2-4 on both sides of each culture dish inlet 2-2-1 are respectively connected to a tension spring I2-6; when the upper ends of the two silo gear rods 2-4 move away from each other in the two V-shaped bar holes, the tension spring I2-6 is pulled apart, and the distance between the two silo gear rods 2-4 increases; when they move in the same direction, the tension spring I2-6 returns to its original state, and the distance between the two silo gear rods 2-4 decreases; each silo support rod 2-3 and two silo gear rods 2-4 form a V-shaped culture dish channel; the two silo gear rods 2-4, one silo support rod 2-3 and two silo guide columns 2-5 are used for positioning the culture dishes.

[0022] like Figure 7As shown, the filling tray 5 includes an upper filling turntable 5-1, a lower filling turntable 5-2, and support columns 5-3; the upper filling turntable 5-1 and the lower filling turntable 5-2 are fixed together by multiple support columns 5-3, and a through hole 5-4 is provided at the center of the upper filling turntable 5-1 and the lower filling turntable 5-2; the upper filling turntable 5-1 and the lower filling turntable 5-2 are both circular disks, and the upper filling turntable 5-1 is composed of two layers of disks set together, and a plurality of culture dish holes 5-1-1 are provided on the lower disk surface, and the plurality of culture dish holes 5-1-1 are provided on the upper surface. The surface is the upper plate culture dish semi-open hole I 5-1-2, the diameter of the culture dish semi-open hole I 5-1-2 is larger than the diameter of the culture dish hole 5-1-1, and a circle of canning plate raised platform 5-1-11 is formed between the culture dish hole 5-1-1 and the culture dish semi-open hole I 5-1-2; a plurality of culture dish semi-open holes II 5-2-1 are provided on the surface of the filling lower turntable 5-2, and the filling plate 5 is divided into area A, area B, area C, area D and area E. The culture dish holes 5-1-1 and the culture dish semi-open holes II 5-2-1 in each area have the same aperture and are concentric. The silo and filling tray are respectively linked by a double-outlet turntable motor, which overcomes the shortcomings of the existing technology that the silo and filling tray are respectively coordinated with the motor gear through the silo and filling tray gears, resulting in errors in the rotation of the silo and filling tray, and poor synchronization between the culture medium out of the silo and the filling tray receiving the culture medium. It can accurately achieve the consistency of the synchronization between the culture medium out of the silo and the filling tray receiving the culture medium; while saving space, it ensures that the opening and closing of the culture dish lid are completed in the minimum range, effectively reducing the risk of contamination.

[0023] like Figure 8 As shown, one end of the silo support plate 4 is a rectangular support plate, and the other end is a semicircular support plate. A culture dish inlet hole 4-1, a culture dish outlet hole 4-2 and two culture dish blocking columns 4-3 are provided on the semicircular support plate surface. A U-shaped culture dish support plate 4-4 is symmetrically fixed on both sides of the culture dish outlet hole 4-2. The front ends of the two U-shaped culture dish support plates 4-4 are exposed in the culture dish outlet hole 4-2, and the two U-shaped culture dish support plates 4-4 can be flipped upward by hinges.

[0024] like Figure 13As shown, the filling peristaltic pump assembly 6 includes a peristaltic stepper motor 6-1, a peristaltic pump 6-2, a peristaltic pump head 6-2-1, an assembly motor bracket 6-3, a pump head fixing seat 6-4, a shock-absorbing column 6-5, an assembly base plate 6-6, and a peristaltic motor sleeve 6-7; the assembly motor bracket 6-3 is fixed to the pump head fixing seat 6-4, and is fixed to the assembly base plate 6-6 through four shock-absorbing columns 6-5 respectively, the peristaltic stepper motor 6-1 is fixed to the assembly motor bracket 6-3, and the peristaltic motor sleeve 6-7 is fixed to the assembly motor bracket 6-3. The peristaltic pump 6-2 and the peristaltic motor sleeve 6-7 are respectively fixed to the pump head fixing seat 6-4. The shaft of the peristaltic stepper motor 6-1 is fixed to the peristaltic pump 6-2 through the assembly motor bracket 6-3, the peristaltic motor sleeve 6-7, the pump head fixing seat 6-4 in sequence; the peristaltic stepper motor 6-1 rotates clockwise, driving the rotor of the peristaltic pump 6-2 to peristalt along the length direction of the pipeline, so that the liquid in the pipeline is transported through the discharge pipe. The peristaltic pump head 6-2-1 is a Y15 peristaltic pump head.

[0025] like Figure 14 、 Figure 15 As shown, the constant temperature filling device 7 includes a filling head bracket 7-1, a filling head 7-2, a heating thermostat 7-3, a heating plate insulation board 7-4 and a thermocouple 7-5; a heating thermostat 7-3 and a thermocouple 7-5 are respectively provided in the open groove 7-1-1 on the lower end surface of the filling head bracket 7-1, the heating plate insulation board 7-4 is fixed on the lower end surface of the filling head bracket 7-1, and the filling head 7-2 is arranged in the U-shaped opening at the upper end of the filling head bracket 7-1; the heating thermostat 7-3 is used to maintain the constant temperature of the filling culture medium to prevent the culture medium from solidifying during filling; the thermocouple 7-5 is used to identify the real-time temperature of the heating thermostat 7-3 and control the heating temperature of the heating thermostat 7-3 through the PLC all-in-one machine; the filling head 7-2 is used for filling the culture medium.

[0026] like Figure 16 As shown, the silo motor assembly 8 includes a double-outlet turntable motor Ⅰ8-1, a disc reducer Ⅰ8-2, a motor fixing sleeve Ⅰ8-3, a motor bracket Ⅰ8-4, a return-to-zero code disc baffle Ⅰ8-5, and a U-shaped sensor Ⅰ8-6; one end of the double-outlet turntable motor Ⅰ8-1 shaft is fixed to the disc reducer Ⅰ8-2, the motor bracket Ⅰ8-4 is fixed to the double-outlet turntable motor Ⅰ8-1, the motor fixing sleeve Ⅰ8-3 is fixed to the disc reducer Ⅰ8-2, and the other end of the double-outlet turntable motor Ⅰ8-1 shaft is fixed to the return-to-zero code disc baffle Ⅰ8-5 Fixed, U-shaped sensor Ⅰ8-6 is fixed on the lower end of motor bracket Ⅰ8-4, and return-to-zero code disk baffle Ⅰ8-5 corresponds to U-shaped sensor Ⅰ8-6; double-outlet turntable motor Ⅰ8-1 rotates clockwise or counterclockwise, driving return-to-zero code disk baffle Ⅰ8-5 to rotate accordingly; disc reducer Ⅰ8-2 is used to decelerate double-outlet turntable motor Ⅰ8-1; return-to-zero code disk baffle Ⅰ8-5 is used to block the light emitted by U-shaped sensor Ⅰ8-6; U-shaped sensor Ⅰ8-6 is used to identify the trigger control of the silo rotation reaching position.

[0027] like Figure 17 As shown, the filling disk motor assembly 9 includes a double-outlet turntable motor II 9-1, a disc reducer II 9-2, a motor fixing sleeve II 9-3, a motor bracket II 9-4, a return-to-zero code disk baffle II 9-5 and a U-shaped sensor II 9-6; the silo motor assembly 8 and the filling disk motor assembly 9 have the same structure, and the installation structure of each component is also the same.

[0028] like Figure 18 、 Figure 19 As shown, the gripper motor assembly 10 includes a gripper stepper motor 10-1, a gripper cam 10-2, a gripper in-position stopper 10-3, and a gripper in-position sensor 10-4. The gripper cam 10-2 is circular, with two arcuate holes 10-2-1 defined on its surface. A gripper in-position stopper 10-3 is secured to its underside. The gripper cam 10-2 is secured to the shaft of the gripper stepper motor 10-1. The gripper cam 10-2 is used to drive the gripper arm to expand or contract. The gripper in-position stopper 10-3 blocks light emitted by the gripper in-position sensor 10-4. The gripper in-position sensor 10-4 detects the triggering of the gripper arm's expansion or contraction. The structural design of the gripper cam and gripper arm enables precise control of the removal of each culture dish from the lower dish channel of the silo.

[0029] like Figure 20 、 Figure 21 As shown, the material receiving motor assembly 11 includes a screw stepper motor Ⅰ11-1, a screw motor bracket Ⅰ11-2, a screw Ⅰ11-3, a screw guide cylinder Ⅰ11-4, a guide cylinder material plate Ⅰ11-5, a screw nut Ⅰ11-6, a motor zero point baffle Ⅰ11-7, a motor PTFE pad Ⅰ11-8, and a U-shaped sensor Ⅲ11-9; the screw stepper motor Ⅰ11-1 is fixed to the bottom plate at the lower end of the screw motor bracket Ⅰ11-2, one end of the screw Ⅰ11-3 passes through the bottom plate of the screw motor bracket Ⅰ11-2 and is connected to the shaft of the screw stepper motor Ⅰ11-1, the other end of the screw Ⅰ11-3 is screwed to the screw nut Ⅰ11-6, one end of the screw guide cylinder Ⅰ11-4 passes through the upper end of the screw motor bracket Ⅰ11-2 and is fixed to the screw nut Ⅰ11-6, and the guide cylinder material plate Ⅰ11 -5 is fixed on the other end of the screw guide cylinder Ⅰ11-4, the motor PTFE pad Ⅰ11-8 is fixed to the side of the screw motor bracket Ⅰ11-2, the screws of the motor zero point baffle Ⅰ11-7 pass through the strip holes of the motor PTFE pad Ⅰ11-8 and the screw motor bracket Ⅰ11-2 in turn, and are fixed on the screw guide cylinder Ⅰ11-4, and the U-shaped sensor Ⅲ11-9 is fixed below the strip hole of the motor PTFE pad Ⅰ11-8; the screw stepper motor Ⅰ11-1 rotates clockwise or counterclockwise, and cooperates with the screw nut Ⅰ11-6 through the screw Ⅰ11-3 to drive the screw guide cylinder Ⅰ11-4 to move up and down; the U-shaped sensor Ⅲ11-9 is used to identify the position of the loading tray rotation; the motor zero point baffle Ⅰ11-7 is used to block the light emitted by the U-shaped sensor Ⅲ11-9.

[0030] like Figure 22 As shown, the ejecting motor assembly 12 includes a screw stepper motor Ⅱ12-1, a screw motor bracket Ⅱ12-2, a screw Ⅱ12-3, a screw guide cylinder Ⅱ12-4, a guide cylinder material plate Ⅱ12-5, a screw nut Ⅱ12-6, a motor zero point baffle Ⅱ12-7, a motor PTFE pad Ⅱ12-8 and a U-shaped sensor Ⅳ12-9; the receiving motor assembly 11 and the ejecting motor assembly 12 have the same structure, and the installation structure of each component is also the same.

[0031] like Figure 23 、 Figure 24 As shown, the culture medium uniformity control device 13 includes a control stepper motor 13-1, a control wheel 13-2, a dust cover 13-2-1, an O-ring 13-3, a control motor plate 13-4, a motor plate slide 13-5, a tension spring II 13-6, and a bolt 13-7; the control motor plate 13-4 is arranged on the two raised slides 13-5-1 on the two sides of the motor plate slide 13-5, and the two raised surfaces 13-4-1 of the control motor plate 13-4 are connected to the two end surfaces of one end of the motor plate slide 13-5. The control stepper motor 13-1 is arranged in the opening at the other end of the motor plate slide 13-5 and is fixed to the control motor plate 13-4. The control wheel 13-2 axle 13-2-2 is fixed to the shaft of the control stepper motor 13-1 through the axle hole 13-4-3. The O-ring 13-3 is set on the control wheel 13-2. The dust cover 13-2-1 is buckled into the dust cover hole 13-2-3. A bolt 13-7 is fixed to the nut I 13-5-2 of the motor plate slide 13-5. The other bolt 13-7 is screwed to the nut II 13-4-2 fixed to the control motor plate 13-4, and the tension spring II 13-6 is hung on the two bolts 13-7; the control stepper motor 13-1 rotates clockwise or counterclockwise, driving the control wheel 13-2 to rotate accordingly; when the control motor plate 13-4 moves on the raised slideways 13-5-1 on both sides of the motor plate slideway 13-5 toward the two raised surfaces 13-4-1, the tension spring II 13-6 is pulled open, and the control motor plate 13 -4's two raised surfaces 13-4-1 are clearance-matched with the two end surfaces at one end of the motor plate slide 13-5, controlling the stepping motor 13-1 and the control wheel 13-2 to move in phase. After the tension spring II 13-6 returns to its original state, the control stepping motor 13-1 and the control wheel 13-2 are restored to their original state, controlling the two raised surfaces 13-4-1 of the motor plate 13-4 to contact and match with the two end surfaces at one end of the motor plate slide 13-5, and the tension spring II 13-6 is used to restore the position of the control wheel 13-2.

[0032] Figure 26 Schematic diagram of the structure of a culture dish, the culture dish 21 is composed of a culture dish cover 21 - 1 and a culture dish bottom 21 - 2 .

[0033] The silo 2 of this inoculation device is equipped with a total of 10 V-shaped culture dish channels, 9 of which are set as lower dish channels and 1 V-shaped culture dish channel is set as a stacking channel. Each lower dish channel can hold 40 culture dishes 21, and the maximum full silo placement can meet the single pouring inoculation task of 360 culture dishes 21.

[0034] Example 1, an automated inoculation method using the pour-over method for microbial counting, wherein the culture medium of this example is plate count agar (PCA); the sample solution of this example is 1 ml of a stock solution of straw mushroom and dark soy sauce added with a 10-fold diluent (phosphate buffer) to prepare a 1:10 dilution.

[0035] This embodiment 1 uses 3 lower dish channels and 1 stacking channel, and 30 culture dishes 21 are placed in each lower dish channel. 1 ml of straw mushroom dark soy sauce stock solution and 10-fold dilution solution are added to each culture dish 21, for a total of 90 culture dishes 21.

[0036] Step 1. Install the safety cover 18 on the main board 1-2 of the table, turn on the machine, light the UV lamp 15-1, and rotate the hopper 2 counterclockwise (when the double-outlet turntable motor Ⅰ8-1 rotates 360 degrees counterclockwise, it drives the hopper 2 to rotate 36 degrees counterclockwise). The stacking channel of the hopper 2 corresponds to the culture dish outlet hole 4-2 of the hopper support plate 4, and the first dish lowering channel corresponds to the culture dish inlet hole 4-1. The hopper 2 stops rotating, and the two gripper arms 14 grippers 14-1 clamp the first culture dish in the first dish lowering channel.

[0037] Step 2: At the dish receiving station, the material receiving motor assembly 11 is started, and the screw stepper motor Ⅰ11-1 drives the guide plate Ⅰ11-5 to move upward, extending out of the culture dish hole 5-1-1 in area A of the filling tray 5, and placed under the culture dish inlet hole 4-1 of the silo support plate 4. The hand motor assembly 10 is started, and the hand stepper motor 10-1 drives the two hand claws 14 through the hand claw cam 10-2 to expand outward, so that the first culture dish falls on the On the guide plate Ⅰ11-5, the claw cam 10-2 drives the two claw arms 14 and the claws 14-1 to retract, clamping the second culture dish. The guide plate Ⅰ11-5 moves downward, and moves the first culture dish down to the culture dish hole 5-1-1 in area A of the filling tray 5. The culture dish cover 21-1 is placed on the raised platform 5-1-11 of the canning tray, and the culture dish bottom 21-2 falls on the table main board 1-2 in the semi-open hole Ⅱ5-2-1 of the culture dish.

[0038] Step 3, inoculation station, the filling tray motor assembly 9 is started, the filling tray 5 rotates counterclockwise (when the double-outlet turntable motor II 9-1 rotates 360 degrees counterclockwise, it drives the filling tray 5 to rotate 72 degrees counterclockwise). When the first culture dish bottom 21-2 is transferred to the inoculation station, the filling tray 5 stops rotating, and the peristaltic stepper motor 6-1 drives the peristaltic pump 6-2 rotor to rotate. The culture medium is continuously pushed in the pipeline. The culture medium is heated to a constant temperature of 50° by the constant temperature filling device 7 constant temperature plate 7-3 and injected into the first culture dish bottom 21-2 through the filling head 7-2. 20 ml of culture medium is injected into each culture dish bottom 21-2. After the culture medium filling is completed, the filling tray 5 swings back and forth rapidly clockwise and counterclockwise. The angle is 10 degrees, the shaking interval is 1 second, the shaking number is 10 times, and the shaking speed is 300 rpm. After the reciprocating shaking, the stepper motor 13-1 is controlled to start, and the O-ring 13-3 of the control wheel 13-2 drives the culture dish to rotate 90 degrees in the filling tray. Due to the centrifugal force generated by the O-ring 13-3 driving the culture dish bottom 21-2 to rotate rapidly in the culture dish semi-open hole Ⅱ5-2-1, the culture medium is evenly attached to the wall, and the control wheel 13-2 stops rotating; at the same time, the second culture dish is transferred to the dish receiving station, and the second culture dish is moved down to the culture dish hole 5-1-1 in area B of the filling tray 5. Repeat step 2 to complete the dish receiving work, and the two gripper arms 14 grippers 14-1 clamp the third culture dish.

[0039] Step 4, transition station, the filling tray motor assembly 9 is started, the filling tray 5 rotates counterclockwise, the first culture dish is transferred to the transition station, the second culture dish is transferred to the inoculation station, the filling tray stops rotating, and step 3 is repeated. After the injection of the culture medium of the second culture dish is completed, the filling tray drives the second culture dish bottom 21-2 and the first culture dish bottom 21-2 to swing back and forth clockwise and counterclockwise at the same time, with a swing angle of 10 degrees, a swing interval of 1 second, a swing number of 10 times, and a swing speed of 300 rpm. The reciprocating swinging stops, and the O-ring of the control wheel drives the second culture dish bottom 21-2 to rotate 90 degrees in the filling tray; at the same time, the third culture dish is transferred to the receiving dish station, and the third culture dish is moved down to the culture dish hole 5-1-1 in area C of the filling tray 5. Step 2 is repeated to complete the receiving dish work, and the two gripper arms 14 grippers 14-1 clamp the fourth culture dish.

[0040] Step 5: At the coding station, the filling plate motor assembly 9 is started, the filling plate 5 rotates counterclockwise, the first culture dish bottom 21-2 is transferred to the printing station and corresponds to the inkjet printer 19 head on the table mainboard 1-2, the second culture dish bottom 21-2 is transferred to the transition station, the third culture dish bottom 21-2 is transferred to the inoculation station, the filling plate 5 stops rotating, and step 3 is repeated. After the injection of the culture medium of the third culture dish is completed, the filling plate drives the third, second and first culture dish bottoms 21-2 to swing back and forth clockwise and counterclockwise at the same time. The shaking angle is 10 degrees, the shaking interval is 1 second, and the shaking times are 10 seconds. The process is as follows: 10 times, the shaking speed is 300 rpm, the reciprocating shaking stops, and the O-ring of the control wheel drives the second culture dish bottom 21-2 to rotate 90 degrees in the filling tray; during the injection of culture medium into the third culture dish, the PLC all-in-one machine controls the inkjet printer 19 to start, and the inkjet printer head completes the coding of the production information of the first culture dish bottom 21-2; at the same time, the fourth culture dish is transferred to the dish receiving station, and the fourth culture dish is moved down to the culture dish hole 5-1-1 in area D of the filling tray 5, and step 2 is repeated to complete the dish receiving work, and the two gripper arms 14 grippers 14-1 clamp the fifth culture dish.

[0041] Step 6, stacking station, the filling tray motor assembly 9 is started, the filling tray 5 rotates counterclockwise, the bottom of the first culture dish is transferred to the stacking station, the bottom of the second culture dish is transferred to the coding station and corresponds to the head of the inkjet printer 19 on the table main board 1-2, the bottom of the third culture dish is transferred to the transition station, the fourth culture dish is transferred to the inoculation station, and the filling tray 5 stops rotating; the top material motor assembly 12 is started, the guide cylinder material plate II 12-5 moves upward, and the bottom 21-2 of the first culture dish is lifted up in the culture dish hole 5-1-1 and fastened with the culture dish cover 21-1, and then the two U-shaped culture dish supporting plates 4-4 in the culture dish outlet hole 4-2 of the silo supporting plate 4 are folded upward and dragged into the stacking channel of the silo 2. After the culture dish is dragged into the stacking channel of the silo 2, the two U-shaped culture dish supporting plates 4-4 return to their original state downward, and the front ends of the two U-shaped culture dish supporting plates 4-4 support the culture dishes in the stacking channel to prevent the culture dishes in the stacking channel from falling and moving downward. ; During the process of dragging the bottom of the first culture dish into the stacking channel of the hopper 2, repeat step three to complete the injection of the culture medium of the fourth culture dish, and the filling tray drives the fourth, third and second culture dish bottoms 21-2 to swing back and forth clockwise and counterclockwise at the same time, with a swing angle of 10 degrees, a swing interval of 1 second, a swing frequency of 10 times, and a swing speed of 300 rpm. When the reciprocating swing stops, the O-ring of the control wheel 13-2 drives the fourth culture dish bottom 21-2 to rotate 90 degrees in the filling tray; During the injection of culture medium for the fourth culture dish, the PLC all-in-one machine controls the inkjet printer 19 to start, and the inkjet printer head completes the coding of the production information of the second culture dish bottom 21-2; At the same time, the fifth culture dish enters the dish receiving station, and the fifth culture dish moves down to the culture dish hole in the filling tray E area, and repeats step two to complete the dish receiving work, and the two gripper arms 14-1 grippers 14-1 clamp the sixth culture dish.

[0042] Step seven, repeat steps two to six, and complete the transfer of the second culture dish bottom to the stacking station and dragging it into the stacking channel, the transfer of the third culture dish bottom to the coding station to complete the coding of the production information, the transfer of the fourth culture dish bottom to the transition station, and the transfer of the fifth culture dish to the inoculation station to complete the injection of culture medium into the culture dish. The filling tray swings back and forth clockwise and counterclockwise, and the O-ring of the control wheel 13-2 drives the fourth culture dish bottom 21-2 to rotate in the filling tray and the receiving, inoculation, transition, coding and stacking processes of the sixth to twenty-fourth culture dishes.

[0043] Step 8. After the 30 culture dishes in the first dish lowering channel of hopper 2 have been moved down, hopper 2 rotates counterclockwise, the first dish lowering channel becomes a stacking channel corresponding to the dish outlet hole 4-2, and the second dish lowering channel corresponds to the dish inlet hole 4-1. Through steps 2 to 6, the 30 culture dishes in the second and third dish lowering channels are received, inoculated, transferred, coded and stacked in sequence.

[0044] The 30 culture dishes in the second dish lowering channel are shaken at an angle of 10 degrees, a shaking interval of 1 second, 15 shaking times, and a shaking speed of 300 rpm in the step three inoculation station; the 30 culture dishes in the third dish lowering channel are shaken at an angle of 10 degrees, a shaking interval of 1 second, 20 shaking times, and a shaking speed of 300 rpm in the step three inoculation station.

[0045] During the inoculation process of the above 90 culture dishes, the UV lamp was always on, thus ensuring the sterile operation of the inoculation area.

[0046] The verification results are as follows: Figure 27 、 Figure 28 and Figure 29 In the figure, the first column on the left shows the results of the test of adding 10-fold dilution of culture medium and straw mushroom dark soy sauce stock solution completed by manual pouring into the plate, coded as scp101-scp105; the second column in the middle and the third column on the right show the results of the test of adding 10-fold dilution of culture medium and straw mushroom dark soy sauce stock solution completed by the inoculation device, coded as cp101-cp115. In the figure, 0.7.10 specifically means, 0.7 is the number, 10 is the number of shaking times 10 times, 15 is the number of shaking times 15 times, and 20 is the number of shaking times 20 times.

[0047] Conclusion: From Figure 27 、 Figure 28 and Figure 29 It was observed that, compared with manual pouring into the plate, the present inoculation device was better than manual pouring into the plate when shaken 15 times and 20 times, and the mixing effect of 20 times was the best.

[0048] From the results of the test on the first column of culture medium manually poured into the flat plate and the addition of 10-fold dilution of the straw mushroom and dark soy sauce stock solution, it can be observed that the addition of 10-fold dilution of the straw mushroom and dark soy sauce stock solution gathered in the center; from the test results of the culture medium completed by the inoculation device in the second column in the middle and the third column on the right and the addition of 10-fold dilution of the straw mushroom and dark soy sauce stock solution, it can be observed that the addition of 10-fold dilution of the straw mushroom and dark soy sauce stock solution was evenly dispersed, with the best degree of mixing. The dilution was fully mixed when there was no clustering.

[0049] Example 2, an automated inoculation method using the pouring method for microbial counting, wherein the culture medium in Example 2 is plate count agar (PCA), and the sample solution in Example 2 is 1 ml of flour added to a 10-fold diluent (phosphate buffer) to prepare a 1:10 dilution. In Example 2, three lower plate channels and one stacking channel are used, with 30 culture dishes 21 placed in each lower plate channel. 1 ml of flour and a 10-fold dilution are added to each culture dish 21, for a total of 90 culture dishes 21. The procedures of connecting, inoculating, transferring, coding and stacking the culture dishes in Example 2 are the same as those in Example 1, except that the parameters are as follows: The first dish channel has 30 culture dishes in the inoculation station at a temperature of 48°, a shaking angle of 20 degrees, a shaking interval of 0.1 seconds, 10 shaking times, and a shaking speed of 250 rpm; The second dish channel has 30 culture dishes in the inoculation station at a temperature of 48 degrees, a shaking angle of 20 degrees, a shaking interval of 0.1 seconds, 15 shaking times, and a shaking speed of 250 rpm. The temperature of 30 culture dishes in the third lower dish channel in the inoculation station is 48 degrees, the shaking angle is 20 degrees, the shaking interval is 0.1 seconds, the shaking number is 20 times, and the shaking speed is 250 rpm.

[0050] The verification results are as follows: Figure 30 、 Figure 31 and Figure 32 In the figure, the first column on the left shows the results of the test of adding 10-fold dilution of culture medium and flour completed by manually pouring into the plate, coded as smp101-smp105; the second column in the middle and the third column on the right show the results of the test of adding 10-fold dilution of culture medium and flour completed by the inoculation device, coded as mp101-mp110. In the figure, 0.7.10 specifically means, 0.7 is the number, 10 is the number of shaking times 10 times, 15 is the number of shaking times 15 times, and 20 is the number of shaking times 20 times.

[0051] Conclusion: From the results of the test of the culture medium and flour added to the 10-fold dilution completed by manually pouring the culture medium into the plate in the first column, it can be observed that the flour is concentrated in the center. From the test results of the culture medium and flour added to the 10-fold dilution completed by the inoculation device in the second column in the middle and the third column on the right, it can be observed that the flour is evenly dispersed and the mixing degree is optimal. The dilution liquid is fully mixed when there is no clustering.

Claims

1. An automated inoculation method for microbial counting by pouring method, characterized in that: Here are the steps: Step 1: Set one V-shaped culture dish channel in the hopper as the stacking channel, and the remaining V-shaped culture dish channels as the lower dish channels. Load multiple culture dishes with sample liquid into the lower dish channels. Turn on the machine, and the first lower dish channel corresponds to the culture dish inlet hole. The two gripper arms clamp the first culture dish in the first lower dish channel. Step 2: At the dish receiving station, the guide plate I moves upward to under the culture dish inlet hole, and the two gripper arms unfold. The first culture dish falls on the guide plate I, and the two gripper arms clamp the second culture dish. The guide plate I moves downward, lowering the first culture dish to the culture dish hole in area A of the filling tray. The culture dish cover rests on the raised platform of the canning tray, and the culture dish bottom falls on the table main board in the semi-opening hole II of the culture dish. Step 3: At the inoculation station, the filling tray rotates counterclockwise, the bottom of the first culture dish is transferred to the inoculation station and stops rotating. The constant temperature culture medium is injected into the bottom of the culture dish through the filling head. After the filling tray swings back and forth clockwise and counterclockwise, the control wheel drives the bottom of the first culture dish to rotate. At the same time, the second culture dish enters the dish receiving station and moves down to the culture dish hole in area B of the filling tray. Repeat step 2 to complete the dish receiving work. The two gripper arms clamp the third culture dish. Step 4: Transfer station. The filling plate rotates counterclockwise, the first culture dish is transferred to the transfer station, and the second culture dish is transferred to the inoculation station. The rotation stops and step 3 is repeated to complete the injection of culture medium into the second culture dish. The filling plate drives the bottom of the second culture dish and the bottom of the first culture dish to rock back and forth clockwise and counterclockwise at the same time. The control wheel drives the bottom of the second culture dish to rotate. At the same time, the third culture dish enters the dish receiving station and moves down to the culture dish hole in area C of the filling tray. Repeat step 2 to complete the dish receiving work. The two gripper arms clamp the fourth culture dish. Step 5: At the coding station, the filling plate rotates counterclockwise, the bottom of the first culture dish is transferred to the coding station, the bottom of the second culture dish is transferred to the transition station, and the bottom of the third culture dish is transferred to the inoculation station. The rotation is stopped and step 3 is repeated to complete the injection of culture medium into the third culture dish. The filling plate drives the bottoms of the third, second, and first culture dishes to swing back and forth clockwise and counterclockwise at the same time. The control wheel drives the bottom of the third culture dish to rotate, and the coding of the bottom of the first culture dish is completed. At the same time, the fourth culture dish enters the dish receiving station and moves down to the culture dish hole in area D of the filling tray. Repeat step 2 to complete the dish receiving work. The two gripper arms clamp the fifth culture dish. Step 6: Stacking station: The filling tray rotates counterclockwise, the bottom of the first culture dish is transferred to the stacking station, the bottom of the second culture dish is transferred to the coding station, the bottom of the third culture dish is transferred to the transition station, and the fourth culture dish is transferred to the inoculation station. The rotation stops, and the guide plate II moves upward, lifting the bottom of the first culture dish and buckling it with the culture dish cover, and then dragging it into the stacking channel of the silo; Repeat step 3 to complete the injection of culture medium into the fourth culture dish. The filling plate drives the fourth, third, and second culture dish bottoms to swing back and forth clockwise and counterclockwise at the same time. The control wheel then drives the fourth culture dish bottom to rotate. The second culture dish bottom is coded. At the same time, the fifth culture dish enters the dish receiving station and moves down to the culture dish hole in area E of the filling tray. Repeat step 2 to complete the dish receiving work. The two gripper arms clamp the sixth culture dish. Step 7, repeating steps 2 to 6, sequentially completing the stacking, coding, stacking, transition, coding, stacking, inoculation, transition, coding, stacking of the second to fifth culture dishes, and the receiving, inoculation, transition, coding, and stacking of the sixth to multiple culture dishes; Step 8: After the multiple culture dishes in the first dish lowering channel of the hopper have finished moving downward, the hopper rotates counterclockwise, and the first dish lowering channel becomes a stacking channel; Through steps 2 to 6, the receiving, inoculation, transfer, coding and stacking of multiple culture dishes in the next dish channel are completed in sequence.

2. The automated inoculation method of pouring method in microbial counting according to claim 1, characterized in that: The clockwise and counterclockwise reciprocating rocking of the filling plate described in step three includes a rocking angle of 1-90 degrees, a rocking speed of 10-500 rpm, a rocking interval of 0.1-3 seconds, and a rocking frequency of 1-20 times; the control wheel drives the bottom of the culture dish to rotate in the filling plate at an angle of 60-90 degrees; and the constant temperature of the culture medium is 36°C-50°C.

3. A vaccination device, characterized in that: A constant temperature filling device, a vertical plate with an ultraviolet lamp, a safety cover and a U-shaped enclosure are fixed on the main plate of the shell table respectively, and a silo supporting plate is fixed on the vertical plate and the U-shaped enclosure. A silo motor assembly and two gripper arms are fixed under the silo supporting plate. The shaft of the silo motor assembly passes through the silo supporting plate and is fixed to the silo arranged on the silo supporting plate, so as to drive the silo to rotate; a filling tray motor assembly, a gripper motor assembly, a material receiving motor assembly, a material ejecting motor assembly, a culture medium uniformity control device and an inkjet printer are fixed under the main plate of the table respectively, and the shaft of the filling tray motor assembly passes through the main plate of the table and is fixed to the filling tray arranged on the main plate of the table, so as to drive the filling tray to rotate; The hand claw cam of the hand claw motor assembly extends out of the table main board and is slidably connected to the two hand claw arms to drive the two hand claw arms to expand outward or retract inward. The guide plate I of the material receiving motor assembly and the guide plate I of the material ejecting motor assembly Ⅱ The control wheels of the culture medium uniformity control device extend out of the main board surface of the table respectively. The control wheels are set in the opening slot of the constant temperature filling device. The control wheels and the filling head guide plate I of the constant temperature filling device correspond to the culture dish inlet hole of the silo support plate. Ⅱ Corresponding to the outlet hole of the culture dish, the inkjet printer head of the inkjet printer is arranged in the inkjet window; the filling peristaltic pump assembly is fixed on the inner bottom plate of the shell, the peristaltic pump head of the filling peristaltic pump assembly is exposed outside the shell panel, and the discharge pipe mouth on the peristaltic pump head is connected with the filling head of the constant temperature filling device; the operation screen, USB data interface and external peristaltic pump interface of the PLC all-in-one are respectively arranged on the shell panel; the PLC all-in-one is respectively connected with the filling peristaltic pump assembly, constant temperature filling device, silo motor assembly, filling tray motor assembly, gripper motor assembly, material receiving motor assembly, top material motor assembly, culture medium uniformity control device, ultraviolet lamp, operation screen, inkjet printer, USB data interface and external peristaltic pump interface.

4. The vaccination device according to claim 3, characterized in that: It also includes a silo shield and a laser sensor. Two laser sensors are fixed on the table main board. The two laser sensors correspond to the filling disks respectively. The two laser sensors are connected to the PLC all-in-one machine. The silo shield is fixed on the silo.

5. The vaccination device according to claim 3, characterized in that: The silo includes a silo lower turntable, a silo upper turntable, a silo support rod, a silo gear rod, a silo guide column, and a tension spring I; the silo upper turntable is an annular disk, and a plurality of culture dish inlets and a plurality of support rod holes I are spaced apart along the annular surface of the annular disk; the silo lower turntable is a circular disk, and an axial hole, a plurality of culture dish half-open outlets, a plurality of support rod holes II, a plurality of silo gear rod holes and a plurality of silo guide column holes are provided on the circular disk; a plurality of the silo guide columns are respectively fixed on a plurality of silo guide column holes, a plurality of the lower ends of the silo support rods are respectively fixed in a plurality of support rod holes II, and a plurality of the upper ends of the silo support rods are respectively fixed in a plurality of In the support rod hole I, the lower ends of the multiple silo gear rods are respectively fixed in the multiple silo gear rod holes, and the upper ends of the multiple silo gear rods extend out of the multiple V-shaped strip holes. The upper ends of the silo gear rods on both sides of the inlet of each culture dish are respectively connected to a tension spring I; when the upper ends of the two silo gear rods move away from each other in the two V-shaped strip holes, the tension spring I is pulled apart, and the distance between the two silo gear rods increases. When moving in the same direction, the tension spring I returns to its original state, and the distance between the two silo gear rods decreases; each silo support rod and the two silo gear rods form a V-shaped culture dish channel; the two silo gear rods, one silo support rod and two silo guide columns are used for positioning the culture dish.

6. The vaccination device according to claim 3, characterized in that: One end of the silo support plate is a rectangular support plate, and the other end is a semicircular support plate. A culture dish inlet hole, a culture dish outlet hole and two culture dish blocking columns are provided on the semicircular support plate surface. A U-shaped culture dish support plate is symmetrically fixed on both sides of the culture dish outlet hole. The front ends of the two U-shaped culture dish support plates are exposed in the culture dish outlet hole, and the two U-shaped culture dish support plates can be flipped upward by hinges.

7. The vaccination device according to claim 3, characterized in that: The filling tray includes an upper filling turntable, a lower filling turntable, and support columns; the upper filling turntable and the lower filling turntable are fixed together by multiple support columns, and a through hole is provided at the center of the upper filling turntable and the lower filling turntable; the upper filling turntable and the lower filling turntable are both circular disks, and the upper filling turntable is composed of two layers of disks set together, and a plurality of culture dish holes are provided on the lower disk surface, and the upper disk culture dish semi-openings are above the multiple culture dish holes. Ⅰ , culture dish holes and culture dish half-open holes Ⅰ A circle of canned dish raised platforms is formed between them; The filling lower turntable is provided with a plurality of culture dish semi-opening holes II, each culture dish hole has the same aperture and is concentric with the corresponding culture dish semi-opening hole II; The filling tray 5 is divided into areas A, B, C, D and E. The culture dish holes and the culture dish half-openings II in each area have the same aperture and are concentric.

8. The vaccination device according to claim 3, characterized in that: The constant temperature filling device includes a filling head bracket, a filling head, a heating constant temperature plate, a heating plate insulation board and a thermocouple; the heating constant temperature plate and the thermocouple are respectively provided in the open groove of the lower end surface of the filling head bracket, the heating plate insulation board is fixed to the lower end surface of the filling head bracket, and the filling head is arranged in the U-shaped opening at the upper end of the filling head bracket; the heating constant temperature plate is used to maintain the constant temperature of the filling culture medium to prevent the culture medium from solidifying during filling; the thermocouple is used to identify the real-time temperature of the heating constant temperature plate, and the heating temperature of the heating constant temperature plate is controlled by the PLC all-in-one machine; the filling head is used for filling the culture medium.

9. The vaccination device according to claim 3, characterized in that: One end of the gripper arm is provided with a bearing, and the other end is a gripper, and the two gripper arms are symmetrically connected to the bottom of the hopper support plate through the finger axis; the gripper motor assembly includes a gripper stepper motor, a gripper cam, a gripper in-position baffle and a gripper in-position sensor, and the gripper in-position sensor is fixed on the table main board, and the gripper stepper motor is fixed under the table main board. The gripper cam connected to the gripper stepper motor extends out of the table main board, and the two arc-shaped strip holes on the gripper cam are respectively mounted on the bearings of the two gripper arms, and the gripper in-position baffle fixed on the gripper cam corresponds to the gripper in-position sensor. The gripper stepper motor rotates clockwise or counterclockwise, and cooperates with the bearings of the two gripper arms through the two arc-shaped strip holes of the gripper cam to drive the two gripper arms to expand outward or contract inward.

10. The vaccination device according to claim 3, characterized in that: The culture medium uniformity control device includes a control stepper motor, a control wheel, a dust cover, an O-ring, a control motor plate, a motor plate slide, a tension spring II, and a bolt; the control motor plate is arranged on the two raised slides on the inner side of the motor plate slide, and the two raised surfaces of the control motor plate are in contact with the two end faces at one end of the motor plate slide. The control stepper motor is arranged in the opening at the other end of the motor plate slide and is fixed to the control motor plate. The control wheel axle is fixed to the shaft of the control stepper motor through the axle hole. The O-ring is sleeved on the control wheel, and the dust cover is buckled in the dust cover hole. One bolt is screwed to the nut I fixed to the motor plate slide, and the other bolt is screwed to the nut II fixed to the control motor plate. The tension spring II is hung on the two bolts; the control stepper motor rotates clockwise or counterclockwise to drive the control wheel to rotate accordingly.

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