CN114107010B - Device and method for automatically and synchronously coating culture dish in multiple areas - Google Patents
Device and method for automatically and synchronously coating culture dish in multiple areas Download PDFInfo
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- CN114107010B CN114107010B CN202111406573.0A CN202111406573A CN114107010B CN 114107010 B CN114107010 B CN 114107010B CN 202111406573 A CN202111406573 A CN 202111406573A CN 114107010 B CN114107010 B CN 114107010B
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
- C12M33/02—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus by impregnation, e.g. using swabs or loops
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/02—Form or structure of the vessel
- C12M23/08—Flask, bottle or test tube
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/48—Holding appliances; Racks; Supports
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
- C12M33/04—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus by injection or suction, e.g. using pipettes, syringes, needles
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/40—Means for regulation, monitoring, measurement or control, e.g. flow regulation of pressure
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/20—Reduction of greenhouse gas [GHG] emissions in agriculture, e.g. CO2
- Y02P60/21—Dinitrogen oxide [N2O], e.g. using aquaponics, hydroponics or efficiency measures
Abstract
The invention discloses a device for automatically and synchronously coating a culture dish in multiple areas, which comprises a main body frame, wherein the main body frame comprises a bottom plate and a top plate, the top plate is provided with a plurality of positioning round holes, the positioning round holes are used for placing a drip tube, the drip tube passes through the positioning round holes, microorganism bacterial suspension is dripped on the culture dish through the drip tube, then a driving device is used for driving a coating device to uniformly coat the microorganism bacterial suspension, and compared with manual coating, the driving device controls the coating device to coat microorganisms, so that the risk of mutual pollution among microorganisms is greatly reduced; in addition, a plurality of drip pipes can be placed in a plurality of locating holes simultaneously, the plurality of drip pipes can be used for synchronously dripping liquid of a plurality of microorganisms, and the coating device can also simultaneously carry out the coating of a plurality of microorganisms and a plurality of areas, so that the time required for coating the culture dish can be greatly reduced.
Description
Technical Field
The invention relates to the field of partitioned culture of microbial culture dishes, in particular to an automatic multi-region synchronous coating culture dish device and method.
Background
In the cultivation of microorganisms, it is often necessary to perform a microorganism plating operation in order to count, isolate and purify the microorganisms. At present, the method for coating the culture dish with the microorganisms mainly depends on manual operation, and only one microorganism bacterial suspension can be coated at a time, so that the operation is time-consuming and labor-consuming. Meanwhile, only one microorganism is usually cultured on one culture dish, which causes waste of resources. There are few documents reporting that different microorganisms are cultivated in a plurality of areas in one culture dish, but the method of coating the culture dish with the microorganisms is dependent on manual operation, and the microorganisms are coated on the culture dish one by one with care, which is easy to cause mutual pollution among different microorganisms and is time-consuming and labor-consuming.
Disclosure of Invention
The invention aims to provide an automatic multi-region synchronous coating culture dish device and a method thereof, which solve the problems that the multi-region coating of microorganisms on the culture dish is easy to pollute and is relatively time-consuming.
The invention is realized by the following technical scheme:
the device comprises a main body frame, wherein the main body frame comprises a bottom plate with a placement part, the placement part is used for fixedly placing the culture dish, the main body frame also comprises a top plate with a plurality of positioning round holes, the positioning round holes are used for placing drip pipes, and the drip pipes are used for dripping liquid to the culture dish; the bottom plate supports the top plate through the fixed supporting piece; the main body frame is also provided with a coating device and a driving device; the driving device is used for driving the coating device to uniformly coat the microbial suspension.
The device realizes automatic multi-region synchronous culture dish coating: the liquid dropping pipe passes through the positioning round hole, the microorganism bacterial suspension is dropped on the culture dish through the liquid dropping pipe, then the driving device is used for driving the coating device to uniformly coat the microorganism bacterial suspension, and the driving device controls the coating device to coat microorganisms, so that the risk of mutual pollution among microorganisms is greatly reduced compared with the manual coating; in addition, a plurality of drip pipes can be placed in a plurality of locating holes simultaneously, the plurality of drip pipes can be used for synchronously dripping liquid of a plurality of microorganisms, and the coating device can also simultaneously carry out the coating of a plurality of microorganisms and a plurality of areas, so that the time required for coating the culture dish can be greatly reduced.
Further, the placing portion is a culture dish groove arranged on the center of the bottom plate, the culture dish groove is matched with the culture dish, and the depth of the culture dish groove is not larger than the height of the side edge of the culture dish. The culture dish recess sets up the position in order to fixed culture dish, and culture dish recess degree of depth is not greater than culture dish side height in order to be more convenient when taking out the culture dish.
Further, the fixed support piece is arranged as two support plates, the distance between the two support plates is larger than the diameter of the culture dish groove, and the two support plates are parallel to each other and are connected with the bottom plate perpendicular to the top plate. The fixed support piece is arranged as the supporting plate, so that more two sides are shielded when the liquid is dripped, and side pollution is prevented.
Further, the locating round holes are evenly distributed around the center circumference of the top disc. The purpose is to accomplish and evenly drip into microorganism fungus suspension on the culture dish, especially when using in multiple microorganism type, evenly distributed's dropping liquid pipe can peg graft on evenly distributed's location round hole to accomplish even coating.
Further, a telescopic mechanism is arranged in the center of the top disc and connected with the coating device, the coating device comprises a connecting rod, one end of the connecting rod is connected with the telescopic mechanism, a hollow cylinder is arranged at the other end of the connecting rod, and a plurality of coating paddles are arranged on the circumference of the hollow cylinder; be provided with the dropping liquid hole on the coating oar, the spacing location round hole position on dropping liquid hole and the roof dish is relative, is provided with the coating recess of opening orientation culture dish on the coating oar. The telescopic mechanism is used for extending and retracting the coating device, and the hollow cylinder is arranged to isolate the central position of the culture dish from a blank area so as to prevent microorganisms from being polluted in the middle area of the culture dish.
Further, the driving device is used for driving the coating device to rotate and driving the coating paddles to rotate on the culture dish according to a set value, so as to uniformly coat the microbial suspension. The driving device is set as a motor, and the forward and reverse rotation of the motor drives the coating device to rotate forward and reverse, so that the coating paddle is driven to rotate on the culture dish according to a set value, and the coating device is used for uniformly coating microbial suspension.
Further, a control system is further arranged on the top disc, a pressure sensing device is arranged between the telescopic mechanism and the coating device, the pressure sensing device is used for sensing the pressure of the end part of the coating device, and the control system is used for receiving signals of the pressure sensing device and controlling the telescopic mechanism to stretch. The pressure sensing device is used for receiving pressure signals, the pressure signals are transmitted to the control system, the control system processes the pressure signals, and the pressure signals are converted and then used for controlling the expansion of the expansion mechanism.
A coating method of an automatic multi-region synchronous coating culture dish device, comprising the following steps:
s1, determining the types of microorganisms and the number of coating areas, and attaching a drip tube and a corresponding coating device; the type and the coating quantity of microorganisms are determined, and the quantity of coating paddles on the corresponding drip tube and the coating device is set again to correspond to the quantity of coating areas one by one.
S2, taking down the culture dish cover, and placing the culture dish in a culture dish groove on the bottom plate;
s3, connecting one end of a high-temperature-resistant hose connected with a peristaltic pump to a bacteria liquid bottle, connecting the other end of the high-temperature-resistant hose to a liquid dropping pipe, starting the peristaltic pump, and pumping the microbial bacterial suspension until the microbial bacterial suspension reaches an experimental set value; the peristaltic pump is used for pumping the microbial suspension in the bacterial liquid bottle.
S4: after the peristaltic pump is stopped, the telescopic mechanism stretches the coating device to the culture dish, and the motor controls the coating device to carry out coating movement through a set coating area and microorganism types;
s5: after the coating movement is finished, the telescopic mechanism contracts the coating device to enable the device to recover to the initial position, and the telescopic mechanism stops working to finish the whole coating work.
Further, the coating device, the liquid dropping pipe, the high-temperature-resistant hose and the fungus liquid bottle all need to be subjected to high-pressure steam sterilization, drying and ultraviolet sterilization before being used, and the steps are completed in an ultra-clean workbench.
Further, the telescopic mechanism is arranged on one side, close to the bottom plate, of the center of the top plate, a pressure sensing device is arranged between the telescopic mechanism and the coating device, and when the bottom of the coating device touches the culture dish, the pressure sensing device senses pressure, so that the telescopic mechanism stops stretching.
Compared with the prior art, the invention has the following advantages and beneficial effects:
the device comprises a main body frame, wherein the main body frame comprises a bottom plate and a top plate, the top plate is provided with a plurality of positioning round holes, the positioning round holes are used for placing a drip tube, the drip tube passes through the positioning round holes, microorganism bacterial suspension is dripped on the culture dish through the drip tube, then a driving device is used for driving a coating device to uniformly coat the microorganism bacterial suspension, and the driving device is used for controlling the coating device to coat microorganisms, so that the risk of mutual pollution among microorganisms is greatly reduced compared with manual coating; in addition, a plurality of drip pipes can be placed in a plurality of locating holes simultaneously, the plurality of drip pipes can be used for synchronously dripping liquid of a plurality of microorganisms, and the coating device can also simultaneously carry out the coating of a plurality of microorganisms and a plurality of areas, so that the time required for coating the culture dish can be greatly reduced.
Drawings
In order to more clearly illustrate the technical solutions of the exemplary embodiments of the present invention, the drawings that are needed in the examples will be briefly described below, it being understood that the following drawings only illustrate some examples of the present invention and therefore should not be considered as limiting the scope, and that other related drawings may be obtained from these drawings without inventive effort for a person skilled in the art. In the drawings:
FIG. 1 is a schematic three-dimensional structure of an entire apparatus according to an embodiment of the present invention;
fig. 2 is a schematic structural diagram of a detailed view of a coating device of the device according to an embodiment of the present invention;
fig. 3 is a schematic structural diagram of a detailed view of a coating blade a of the device according to an embodiment of the present invention;
FIG. 4 is a schematic diagram showing the distribution of four areas of the same microorganism in a culture dish synchronously coated according to an embodiment of the present invention;
FIG. 5 is a schematic diagram showing the distribution of four areas of synchronous coating of the same microorganism concentration gradients in a culture dish according to the embodiment of the present invention;
FIG. 6 is a schematic view showing the distribution of three areas of three microorganisms in a culture dish synchronously coated according to an embodiment of the present invention;
FIG. 7 is a schematic diagram showing the distribution of the same microorganism in a culture dish with different areas coated synchronously, wherein the areas are one area, two areas, three areas and four areas in sequence from left to right;
fig. 8 is a schematic front view of the whole device according to the embodiment of the present invention.
In the drawings, the reference numerals and corresponding part names:
1-bottom plate, 2-culture dish recess, 3-backup pad, 4-top tray, 5-location round hole, 6-control system, 7-time switch, 8-contactless switch, 9-motor, 10-telescopic machanism, 11-pressure sensing device, 12-coating device, 1201-coating oar, 1202-hollow cylinder, 1203-connecting rod, 1204-coating recess, 1205-drip hole, 13-drip pipe, 14-high temperature resistant hose, 15-peristaltic pump, 16-fungus liquid bottle.
Detailed Description
For the purpose of making apparent the objects, technical solutions and advantages of the present invention, the present invention will be further described in detail with reference to the following examples and the accompanying drawings, wherein the exemplary embodiments of the present invention and the descriptions thereof are for illustrating the present invention only and are not to be construed as limiting the present invention.
Example 1
1-3, an automatic multi-region synchronous culture dish coating device comprises a main body frame, wherein the main body frame comprises a bottom plate 1 with a placing part, the placing part is used for fixedly placing a culture dish, the main body frame also comprises a top plate 4 with a plurality of positioning round holes 5, the positioning round holes 5 are used for placing a drip tube 13, and the drip tube 13 is used for dripping liquid to the culture dish; the bottom plate 1 supports the top plate 4 through a fixed support; the main body frame is also provided with a coating device 12 and a driving device; the driving device is used for driving the coating device 12 to uniformly coat the microbial suspension.
The device realizes automatic multi-region synchronous culture dish coating: the drip tube 13 passes through the positioning round hole 5, the microorganism bacterial suspension is dripped on the culture dish through the drip tube 13, then the driving device is used for driving the coating device 12 to uniformly coat the microorganism bacterial suspension, and compared with manual coating, the driving device controls the coating device 12 to coat microorganisms, so that the risk of mutual pollution among microorganisms is greatly reduced; in addition, a plurality of drip pipes 13 can be simultaneously placed in a plurality of positioning holes, a plurality of drip pipes 13 can be used for synchronously dripping various microorganisms, the coating device 12 can also simultaneously carry out the coating of various microorganisms and multiple areas, and the time required for coating the culture dish can be greatly reduced.
The bottom plate 1 is used for accommodating a culture dish and fixing two support plates 3; the culture dish groove 2 on the bottom plate 1 is positioned at the center of the bottom plate 1, the culture dish groove 2 is consistent with the culture dish in shape and is cylindrical, and the central axis of the culture dish groove 2 is positioned in the same vertical line with the central axes of the coating device 12, the pressure sensing device 11, the telescopic mechanism 10, the motor 9 and the top plate 4; the support piece preferably uses two support plates 3 which are respectively erected on two sides of the culture dish groove 2 on the bottom plate 1, have the same distance with the center point of the culture dish groove 2, are mutually parallel, and are respectively fixedly connected with the top plate 4 and the bottom plate 1 from top to bottom, and are used for fixing the top plate 4 and the bottom plate 1; the top disc is a circular plate, a plurality of positioning round holes 5 which take the center point of the top disc 4 as the center of a circle and are uniformly distributed at equal distance are formed in the inner part of the top disc with a certain distance from the two support plates 3, the aperture of each positioning round hole 5 is smaller than the top of the drip tube 13 and is used for containing the drip tube 13, the distance between every two positioning round holes 5 on the same diameter of the top disc 4 is not more than the diameter of the culture dish groove 2, and the drip tube 13 can be ensured to drip microorganism bacterial suspension onto the culture dish after being installed;
the portion of placing is for setting up the culture dish recess 2 on bottom plate 1 center, and culture dish recess 2 and culture dish adaptation, and culture dish recess 2 degree of depth is not greater than culture dish side height. The dish groove 2 is provided for fixing the position of the dish, and the depth of the dish groove 2 is not greater than the height of the side edge of the dish for convenience in taking out the dish.
The fixed support is arranged as two support plates 3, the distance between the two support plates 3 is larger than the diameter of the culture dish groove 2, and the two support plates 3 are parallel to each other and are connected with the bottom plate 1 in a way of being perpendicular to the top plate 4. The fixed support is set to the backup pad 3 effect is when carrying out the dropping liquid, and both sides shelter from more, prevent the side pollution.
The positioning round holes 5 are uniformly distributed around the center circumference of the top disk 4. The purpose is to finish the uniform dripping of the microbial suspension on the culture dish, especially when being used for various microbial species, the uniformly distributed drip tube 13 can be inserted into the uniformly distributed positioning round holes 5, thereby finishing the uniform coating.
The center of the top disk 4 is provided with a telescopic mechanism, the telescopic mechanism is connected with the coating device 12, the coating device 12 comprises a connecting rod 1203, one end of the connecting rod 1203 is connected with the telescopic mechanism 10, the other end of the connecting rod 1203 is provided with a hollow cylinder 1202, and a plurality of coating paddles 1201 are arranged on the circumference of the hollow cylinder 1202; the coating paddle 1201 is provided with a liquid dropping hole 1205, the liquid dropping hole 1205 is opposite to the limit positioning round hole 5 on the top disk 4, and the coating paddle 1201 is provided with a coating groove 1204 with an opening facing the culture dish. The telescoping mechanism is used to extend and retract the applicator 12, and the hollow cylinder 1202 is provided to isolate the center of the dish from the empty space and prevent contamination of microorganisms in the middle area of the dish.
The coating device 12 is composed of a connecting rod 1203 with a threaded top, a hollow cylinder 1202 with a downward opening at the bottom, and a coating paddle 1201 which are fixedly connected. The screw thread at the top of the connecting rod 1203 is used for connecting the coating device 12 with the pressure sensing device 11 in a screw thread manner; the hollow cylinder 1202 with the downward opening at the bottom is fixedly connected with the unthreaded end of the connecting rod 1203, coating paddles 1201 are uniformly distributed on the circumference of the hollow cylinder 1202, and the hollow cylinder 1202 with the downward opening is used for forming a round blank area on the center of the culture medium so as to prevent microorganisms in different areas from polluting each other; uniformly distributed coating paddles 1201 are used for microbial coating of different areas on the medium. For example: the number of coating paddles 1201 of the coating device 12 can be n (n is equal to or greater than 2 and is an integer), and synchronous coating of a plurality of areas on one culture dish is realized by replacing the coating devices 12 with different numbers of coating paddles 1201. The coating blade 1201 of the coating device 12 rotates by X ° (0+.x+.365), determined by how many areas are coated, X ° being 360 ° when there are only 1 coating area; when there are n (n.gtoreq.2, and n is an integer) coating zones, then X is 360/n-5. In order to prevent contamination, the angle of the blank area may be increased according to the kind of microorganism.
The telescopic mechanism 10 is connected to the right below the motor 9 for the up-and-down movement of the pressure sensing device 11 and the coating device 12 in the vertical direction. When coating is needed, the telescopic mechanism 10 stretches, so that the coating device 12 moves downwards to contact with the culture medium on the culture dish, and the pressure sensing device 11 senses certain pressure, so that the telescopic mechanism 10 stops stretching; after the coating is completed, the telescopic mechanism 10 is contracted, and the pressure sensing device 11 and the coating device 12 are lifted up until the starting position is restored.
The driving device is used for driving the coating device 12 to rotate, driving the coating paddle 1201 to rotate on the culture dish according to a set value, and uniformly coating the microbial suspension. The driving device is set as a motor 9, and the forward and reverse rotation of the motor 9 drives the coating device 12 to rotate forward and reverse, so as to drive the coating paddle 1201 to rotate on the culture dish according to a set value for uniformly coating the microbial suspension. The motor 9 is embedded and fixed at the center point of the top disc 4, and passes through the top disc 4 to be connected with the telescopic mechanism 10. The device is used for driving the telescopic mechanism 10, the pressure sensing device 11 and the coating device 12 to perform forward and reverse rotation at a certain angle so as to realize synchronous coating of microorganisms on the culture dish in multiple areas; the motor 9 is provided with a timing switch 7 and a non-contact switch 8, and the timing switch 7 is positioned above the motor 9 and used for controlling the rotation time of the motor 9 so as to stop the coating operation after a certain time of the coating operation is realized; a contactless switch 8 is located above the motor 9 for controlling the forward and reverse rotation of the motor 9.
The top disk 4 is also provided with a control system 6, a pressure sensing device 11 is arranged between the telescopic mechanism 10 and the coating device 12, the pressure sensing device 11 is used for sensing the pressure of the end part of the coating device 12, and the control system 6 is used for receiving signals of the pressure sensing device 11 and controlling the telescopic mechanism 10 to stretch and retract. The pressure sensing device 11 is configured to receive a pressure signal, transmit the pressure signal to the control system 6, process the pressure signal by the control system 6, and convert the signal to control the expansion of the expansion mechanism 10.
The pressure sensing device 11 is connected to the right lower part of the telescopic mechanism 10 and is used for sensing the pressure of the culture medium on the culture dish when the coating device 12 moves downwards, so as to control the telescopic mechanism 10 to stop stretching. An internal thread round hole is arranged right below the pressure sensing device 11 and is used for the threaded connection of the top of the connecting rod 1203 of the coating device 12; the control system 6 is fixed on the top disk 4 and is in the same vertical plane with one of the support plates 3, and is used for controlling the actions and the operations of the contactless switch 8, the motor 9, the telescopic mechanism 10 and the pressure sensing device 11.
A coating method of an automatic multi-region synchronous coating culture dish device, comprising the following steps:
s1, determining the type of microorganisms and the number of coating areas, and attaching a drip tube 13 and a corresponding coating device 12; the type and the coating quantity of the microorganisms are determined, and the number of coating paddles 1201 on the corresponding drip tube 13 and the coating device 12 are arranged again to correspond to the number of coating areas one by one.
The drip tube 13 is a hollow stainless steel tube, the bottom end is in a needle point shape, the diameter of the middle part is smaller than the diameter of the positioning round hole 5, and the top end is provided with a small section of tube diameter which is larger than the diameter of the positioning round hole 5, so that the bottom end and the middle part of the drip tube 13 can pass through the positioning round hole 5 and are hung on the top disc 4. The drip tube 13 is used for dripping the microbial suspension which is conveyed by the high temperature resistant hose 14 and is originally filled in the fungus liquid bottle 16 onto the culture medium. The microbial suspension in the fungus liquid bottle 16 is dripped on different areas of the culture medium through the drip pipes 13 with different conveying routes to the different positioning round holes 5. For example: when the number of the batch of coating drip dispensing pipes 13 is 1, 1 positioning round hole 5 can be arbitrarily selected; when the number of the coating drip pipes 13 in the batch is more than or equal to 2, the drip pipes 13 are required to be inserted into the corresponding positioning round holes 5 in a pairwise equidistant manner.
Wherein the fungus liquid bottle 16 is a high temperature resistant bottle with a positioning round hole 5 sealing cover at the top, and is used for containing microorganism fungus suspension. The high temperature resistant hose 14 can extend into the bacteria liquid bottle 16 through the top positioning round hole 5 to convey the microbial bacterial suspension. When different microbial suspensions need to be coated, corresponding bacteria liquid bottles 16 can be added to contain the bacteria liquid.
S2, taking down the culture dish cover, and placing the culture dish in a culture dish groove on the bottom plate;
s3, connecting one end of a high-temperature-resistant hose 14 connected with a peristaltic pump 15 to a bacteria liquid bottle 16, connecting the other end of the high-temperature-resistant hose to a liquid dropping pipe 13, starting the peristaltic pump 15, and pumping the microbial bacterial suspension until the microbial bacterial suspension reaches an experimental set value, and stopping the peristaltic pump 15; peristaltic pump 15 is used to draw the bacterial fluid from bacterial fluid bottle 16.
Wherein the high temperature resistant hose 14 is a hollow hose, one end of the hose extends into the bacteria liquid bottle 16, the middle of the hose passes through the peristaltic pump 15, and the other end of the hose is sleeved at the top of the liquid dropping tube 13. The high temperature resistant hose 14 is used to transport the microbial suspension in the bacteria bottle 16. When different microorganisms need to be conveyed, corresponding high-temperature resistant hoses 14 and drip pipes 13 at different positions can be added and connected in a one-to-one correspondence mode, so that different microorganisms can be dripped on different areas of the culture medium.
Peristaltic pump 15 is a multichannel peristaltic pump 15 and has different flow delivery functions. The microbial bacteria suspension in the bacteria liquid bottle 16 is pumped into the high temperature resistant hose 14, and finally is dripped onto the culture medium through the high temperature resistant hose 14 and the drip tube 13. The flow rate of the peristaltic pump 15 for conveying the microbial suspension is determined according to the concentration of the microbial suspension in the microbial suspension bottle 16 or the size of the area on the culture medium, and when the flow rate reaches a set value, the peristaltic pump 15 stops working. When different microbial suspensions need to be conveyed, the high-temperature-resistant hose 14 can be added, and different high-temperature-resistant hoses 14 can pass through different channels of the peristaltic pump 15, so that conveying of different microbial suspensions is realized. For example: when the number of the drip tubes 13 coated in the batch is n (n is not less than 1 and n is an integer), the peristaltic pump 15 stops working when the microbial suspension dropped out from each drip tube 13 is 200/n microliter.
S4: after the peristaltic pump 15 is stopped, the telescopic mechanism stretches the coating device 12 to the culture dish, and the motor 9 controls the coating device 12 to perform coating movement through a set coating area and microorganism types;
s5: after the coating movement is completed, the telescopic mechanism contracts the coating device 12 to restore the initial position, and the telescopic mechanism stops working to complete the whole coating work.
The coating device 12, the drip tube 13, the high temperature resistant hose 14 and the bacteria liquid bottle 16 are all required to be subjected to high pressure steam sterilization, drying and ultraviolet sterilization before use, and the steps are all completed in an ultra-clean workbench.
The telescopic mechanism is arranged on one side, close to the bottom plate 1, of the center of the top plate 4, a pressure sensing device 11 is arranged between the telescopic mechanism 10 and the coating device 12, and when the bottom of the coating device 12 touches the culture dish, the pressure sensing device 11 senses pressure, so that the telescopic mechanism 10 stops stretching.
The principle of the whole device is as follows: the device comprises a device body frame, a control system 6 and a drive system. The device main body frame comprises a bottom plate 1 and a culture dish groove 2 in the upper center of the bottom plate, two support plates 3 are uniformly distributed on the bottom plate 1 and are arranged on two sides of the culture dish groove 2, and the two support plates 3 are fixedly connected with a top plate 4 and the bottom plate 1 from top to bottom respectively. The top disk 4 is provided with round positioning round holes 5 which are uniformly distributed at equal distance, and the positioning round holes 5 are used for hanging and placing the drip tube 13; one end of a high-temperature-resistant hose 14 extends into the bacteria liquid bottle 16 through a connecting hole at the top of the bacteria liquid bottle 16, the other end of the high-temperature-resistant hose 14 passes through a peristaltic pump 15 and is connected with the top of the liquid dropping pipe 13, and bacteria liquid enters the liquid dropping pipe 13 through the high-temperature-resistant hose 14 and then drops onto a culture dish; the control system 6 is arranged on the top disc 4 at the same vertical line position with the supporting plate 3, so that the control system 6 can be arranged to enable the weight of the arrangement system to find a bearing point; the motor 9 is inlaid in the center position of the top disc 4, and a timing switch 7 and a contactless switch 8 are arranged on the upper portion of the motor 9. The timing switch 7 is used for controlling the rotation time of the motor 9 so as to stop the coating operation after a certain time of the coating operation; the contactless switch 8 is used for controlling the forward and reverse rotation of the motor 9, and the forward and reverse rotation of the motor 9 can drive the forward and reverse rotation of the coating device 12, so that the coating operation is completed. The lower part of the motor 9 is connected with a telescopic mechanism 10, a pressure sensing device 11 is connected under the telescopic mechanism 10, a screw hole is formed under the pressure sensing device 11 and is used for being in threaded connection with a coating device 12, the telescopic mechanism 10 is used for controlling the expansion and contraction of the coating device 12, a control signal is transmitted to a control system 6 through the pressure of the end part of the pressure sensing device 11 sensing the coating device 12, and the expansion and contraction of the telescopic mechanism 10 are controlled.
The principle of the coating device 12 is: the coating paddle 1201 is a downward opening groove, and a cylindrical drip hole 1205 is formed at the top of the groove, the left side plate of the groove is a left coating plate, the front side plate is a front baffle plate, and the right side plate is a right coating plate to form a coating frame, and the tail of the coating frame is connected with the hollow cylinder 1202 of the coating device 12. A small semi-cylindrical bulge is arranged at the uppermost part of the threaded hole at the lower part of the pressure sensing device 11, and a semi-cylindrical notch corresponding to the bulge is arranged at the top of the connecting rod 1203 of the coating device 12 and can be mutually matched. When the screw thread at the top of the coating device 12 is screwed into the screw hole at the lower part of the pressure sensing device 11, the notch at the top of the coating device 12 collides with the semi-cylindrical protrusion, so that the drip hole 1205 of the coating blade 1201 is positioned at the same vertical position as the drip tube 13 suspended directly above. The microbial suspension dropped from the drip tube 13 falls onto the culture medium of the dish through the drip hole 1205 of the coating paddle 1201, and at this time, the microbial suspension on the culture medium is in the space surrounded by the coating frame of the coating paddle 1201 and the hollow cylinder 1202, so as to prevent the microbial suspensions in different areas from contaminating each other during coating. When the coating device 12 rotates by X ° clockwise, the microbial suspension is coated by the left coating plate of the coating frame at this time; when the coating device 12 is rotated counterclockwise by X °, the microbial suspension is coated by the right coating plate of the coating frame. The above steps are repeated for a plurality of times, and after a period of time, the timing switch 7 is started, and the motor 9 stops rotating. The coating blade 1201 of the coating device 12 rotates by X ° (0+.x+.365), determined by how many areas are coated, X ° being 360 ° when there are only 1 coating area; when there are n (n.gtoreq.2, and n is an integer) coating zones, then X is 360/n-5.
Example 2
As shown in fig. 4, the coating method of an automatic multi-zone synchronous coating culture dish device is exemplified by four-zone synchronous coating of same microorganism in one culture dish:
1. the coating device 12, the drip tube 13, the high temperature resistant hose 14 and the bacteria liquid bottle 16 are wrapped with newspaper, and the bacteria liquid bottle is sterilized by high pressure steam at 121 ℃ for 20min. And (3) sterilizing well, and placing the materials in a 60 ℃ oven for drying for standby.
2. The sterilized drip tube 13, the high temperature resistant hose 14, the bacteria liquid bottle 16 and other parts of the device are all placed in an ultra clean bench, and ultraviolet sterilization is performed for 30min.
3. The ultra-clean bench is opened, and the following operations are performed in the ultra-clean bench.
4. The microbial suspension is added into the bacteria liquid bottle 16 through a small hole on the bottle cover of the bacteria liquid bottle 16. The coating device 12 is rotated by the top screw into the threaded hole below the pressure sensing device 11. The tips of the four drip pipes 13 penetrate through four equidistant positioning round holes 5 on the top disk 4 and are hung on the top disk 4. One end of each of the four high-temperature-resistant hoses 14 extends into the bottom of the fungus liquid bottle 16, the other end of each of the four high-temperature-resistant hoses 14 is sleeved on the tops of the four liquid dropping pipes 13, and the middle parts of the four high-temperature-resistant hoses 14 are wound into the peristaltic pump 15.
5. The culture dish with the existing solid culture medium is opened to a top cover and placed in the culture dish groove 2.
6. The peristaltic pump 15 is started, and the microbial suspension in the bacteria liquid bottle 16 is dripped onto four equidistant areas of the culture medium along four high-temperature-resistant hoses 14 and through four drip pipes 13. Peristaltic pump 15 is stopped when each drip tube 13 drips 50 microliters of microbial suspension.
7. The control system 6 is started to control the extension of the telescopic mechanism 10, and when the bottom of the coating device 12 touches the culture medium, the pressure sensing device 11 senses pressure, so that the extension of the telescopic mechanism 10 is stopped. Then the motor 9 is started, the coating device 12 is driven to rotate by 85 degrees clockwise to leave partial blank 5 degrees, the area of the blank area can be adjusted according to different types of requirements of microorganisms, and the microorganisms in different areas are prevented from being polluted; the non-contact switch 8 is started, so that the motor 9 drives the coating device 12 to rotate by 85 degrees anticlockwise, the non-contact switch 8 is started again, and drives the coating device 12 to rotate by 85 degrees clockwise again, and the process is repeated for a plurality of times, so that the four coating paddles 1201 of the coating device 12 uniformly coat the microorganism bacterial suspension dropped on the culture medium on four areas of the culture medium. After a certain time, the timing switch 7 is started, and the motor 9 stops rotating.
8. The telescopic mechanism 10 is shortened, the pressure sensing device 11 and the coating device 12 are restored to the initial positions, and the telescopic mechanism 10 stops working.
9. And taking the coated culture dish out of the culture dish groove 2, and finishing the coating operation.
What should be explained is:
(1) If the microbial suspension is required to be coated on a plurality of culture dishes, a new culture dish is replaced, and the operation is repeated.
(2) If the number of the microorganism-coated areas needs to be changed, the coating device 12, the high-temperature-resistant hose 14 and the liquid dropping tube 13 with the corresponding number of coating paddles 1201 need to be replaced, and the above operations are repeated.
(3) If another microorganism bacterial suspension is to be coated on another culture dish, the coating device 12, the drip tube 13, the high temperature resistant hose 14 and the bacterial liquid bottle 16 are replaced, and the above operations are repeated.
Example 3
As shown in fig. 5, based on embodiment 2, this embodiment provides: four concentration gradients of homologous microorganisms in one culture dish are synchronously coated.
1. A sterilized drip tube 13, a high temperature resistant hose 14 and a fungus liquid bottle 16 are a set of microorganism and fungus suspension conveying system, four sets of microorganism and fungus suspension conveying systems are taken, a coating device 12 with four coating paddles 1201 is arranged, and four concentration gradient synchronous coatings of the same microorganism in a culture dish are completed according to the following operation.
2. In the ultra clean bench, four bacteria liquid bottles 16 are respectively filled with the dilution gradient of 10 -3 、10 -4 、10 -5 、10 -6 Is a microbial suspension of the above-mentioned microorganism. Four drip tubes 13 are inserted into the positioning round holes 5 in a pairwise equidistant manner, and then the device is connected.
3. The peristaltic pump 15 is started, and the microbial suspension in the four bacteria liquid bottles 16 is respectively dripped onto four different areas of the culture medium through the drip pipes 13 along the four high-temperature-resistant hoses 14. When the microbial suspension dropped from each drip tube 13 was 200. Mu.l, the peristaltic pump 15 was stopped.
7. The control system 6 is started to control the extension of the telescopic mechanism 10, and when the bottom of the coating device 12 touches the culture medium, the pressure sensing device 11 senses pressure, so that the extension of the telescopic mechanism 10 is stopped. Then the motor 9 is started, the coating device 12 is driven to rotate by 85 degrees clockwise, the non-contact switch 8 is started, the motor 9 drives the coating device 12 to rotate by 85 degrees anticlockwise, and the non-contact switch is openedThe switch 8 is started again, and the coating device 12 is driven to rotate by 85 degrees clockwise again, so that the coating paddle 1201 of the coating device 12 drops on the culture medium to be 10 times -3 、10 -4 、10 -5 、10 -6 The diluted microbial suspensions of (2) are uniformly spread on four different areas of the medium, respectively. After a certain time, the timing switch 7 is started, and the motor 9 stops rotating.
8. The telescopic mechanism 10 is shortened, the pressure sensing device 11 and the coating device 12 are restored to the initial positions, and the telescopic mechanism 10 stops working.
9. And taking the coated culture dish out of the culture dish groove 2, and completing four concentration gradient synchronous coating operations of the same microorganism in one culture dish.
Example 4
As shown in fig. 6, based on embodiment 2, this embodiment provides: three areas of three microorganisms in one culture dish are coated synchronously.
1. A sterilized drip tube 13, a high temperature resistant hose 14 and a fungus liquid bottle 16 are a set of microorganism fungus suspension conveying system, three sets of microorganism fungus suspension conveying systems are taken, and a coating device 12 with three coating paddles 1201 is configured to complete synchronous coating of three areas of three microorganisms in a culture dish according to the following operation.
2. In the ultra clean bench, three bacterial liquid bottles 16 are filled with microbial bacterial suspensions of the microorganism a, the microorganism b and the microorganism c, respectively. Three drip tubes 13 are inserted into the positioning round holes 5 in pairs at equal distance, and then the device is connected according to fig. 1.
3. The peristaltic pump 15 is started, and the microbial suspension in the three bacteria liquid bottles 16 is respectively dripped onto three different areas of the culture medium through the dripping pipes 13 along the three high-temperature-resistant hoses 14. Peristaltic pump 15 is stopped when each drip tube 13 drips 67 microliters of microbial suspension.
7. The control system 6 is started to control the extension of the telescopic mechanism 10, and when the bottom of the coating device 12 touches the culture medium, the pressure sensing device 11 senses pressure, so that the extension of the telescopic mechanism 10 is stopped. Then the motor 9 is started, the coating device 12 is driven to rotate 115 degrees clockwise, the non-contact switch 8 is started, the motor 9 drives the coating device 12 to rotate 115 degrees anticlockwise, the non-contact switch 8 is started again, the coating device 12 is driven to rotate 115 degrees clockwise again, and the process is repeated for a plurality of times, so that the coating paddles 1201 of the coating device 12 uniformly coat microbial suspensions of the microorganisms a, b and c on the culture medium respectively on three different areas of the culture medium. After a certain time, the timing switch 7 is started, and the motor 9 stops rotating.
8. The telescopic mechanism 10 is shortened, the pressure sensing device 11 and the coating device 12 are restored to the initial positions, and the telescopic mechanism 10 stops working.
9. The coated culture dish is taken out of the culture dish groove 2, and three areas of three microorganisms in one culture dish are synchronously coated.
Example 5:
as shown in fig. 7, based on embodiment 2, this embodiment provides: different numbers of simultaneous coating of different areas of the same microorganism.
1. A sterilized drip tube 13, a high temperature resistant hose 14 and a bacteria liquid bottle 16 are a set of microorganism bacteria liquid conveying system, one set, two sets, three sets and four sets of microorganism bacteria suspension conveying systems are respectively taken, and coating devices 12 with two, three and four coating paddles 1201 are respectively configured, and the number of different areas of microorganisms are coated in batches according to the following operation.
2. In the ultra clean bench, the bacteria liquid bottles 16 are filled with microorganism bacterial suspension, the drip pipes 13 are inserted into the positioning round holes 5 (1 positioning round hole 5 can be selected arbitrarily when the number of the batch of coating drip pipes 13 is 1, and the drip pipes 13 need to be inserted into the corresponding positioning round holes 5 in a pairwise equidistant manner when the number of the batch of coating drip pipes 13 is more than or equal to 2), and then the device is connected according to the figure 1.
3. The peristaltic pump 15 is started, and the microbial suspension in the bacteria liquid bottle 16 is dripped onto the culture medium along the high-temperature-resistant hose 14 through the drip tube 13. When the batch is coated with one, two, three and four drip tubes 13, the peristaltic pump 15 stops working when the microbial suspension dropped from each drip tube 13 is 200 microliter, 100 microliter, 67 microliter and 50 microliter.
7. The control system 6 is started to control the extension of the telescopic mechanism 10, and when the bottom of the coating device 12 touches the culture medium, the pressure sensing device 11 senses pressure, so that the extension of the telescopic mechanism 10 is stopped. Then the motor 9 is started, the coating device 12 is driven to rotate for X degrees clockwise, the non-contact switch 8 is started, the motor 9 drives the coating device 12 to rotate for X degrees anticlockwise, the non-contact switch 8 is started again, the coating device 12 is driven to rotate for X degrees clockwise again, and the process is repeated for a plurality of times, so that the coating paddles 1201 of the coating device 12 uniformly coat the microbial suspension dropped on the culture medium. When the coating areas are 1, 2, 3 and 4, respectively, then X ° is 360 °, 175 °, 115 ° and 85 °, respectively. After a certain time, the timing switch 7 is started, and the motor 9 stops rotating.
8. The telescopic mechanism 10 is shortened, the pressure sensing device 11 and the coating device 12 are restored to the initial positions, and the telescopic mechanism 10 stops working.
9. And taking the coated culture dish out of the culture dish groove 2, and finishing the coating operation.
It should be explained that: the above embodiments illustrate the white portion as sterile blank medium and the shaded portion as microorganism coated area.
The foregoing description of the embodiments has been provided for the purpose of illustrating the general principles of the invention, and is not meant to limit the scope of the invention, but to limit the invention to the particular embodiments, and any modifications, equivalents, improvements, etc. that fall within the spirit and principles of the invention are intended to be included within the scope of the invention.
Claims (7)
1. The device for automatically and synchronously coating the culture dish in multiple areas is characterized by comprising a main body frame, wherein the main body frame comprises a bottom plate (1) with a placement part, the placement part is used for fixedly placing the culture dish, the main body frame further comprises a top plate (4) with a plurality of positioning round holes (5), the positioning round holes (5) are used for placing a drip tube (13), and the drip tube (13) is used for dripping liquid to the culture dish; the bottom plate (1) supports the top plate (4) through a fixed support piece;
the main body frame is also provided with a coating device (12) and a driving device;
the driving device is used for driving the coating device (12) to uniformly coat the microbial suspension;
wherein the positioning round holes (5) are uniformly distributed around the center circumference of the top disk (4);
the center of the top disc (4) is provided with a telescopic mechanism (10), the telescopic mechanism (10) is connected with the coating device (12), the coating device (12) comprises a connecting rod, one end of the connecting rod is connected with the telescopic mechanism (10), the other end of the connecting rod is provided with a hollow cylinder (1202), a plurality of coating paddles (1201) are arranged on the circumference of the hollow cylinder (1202), and the driving device is used for driving the coating paddles (1201) to rotate;
the coating paddle (1201) is provided with a liquid dropping hole (1205), the liquid dropping hole (1205) is opposite to a limit positioning round hole (5) on the top disc (4), and the coating paddle (1201) is provided with a coating groove (1204) with an opening facing the culture dish.
2. The automatic multi-region synchronous culture dish coating device according to claim 1, wherein the placing part is a culture dish groove (2) arranged on the center of the bottom plate (1), the culture dish groove (2) is matched with the culture dish, and the depth of the culture dish groove (2) is not larger than the height of the side edge of the culture dish.
3. Device for the automatic multizone simultaneous coating of petri dishes according to claim 1, characterised in that the fixed support is provided as two support plates (3), the distance between two support plates (3) being larger than the diameter of the dish recess (2), and the two support plates (3) being parallel to each other and being connected perpendicular to the top plate (4) and the bottom plate (1).
4. The automatic multi-region synchronous culture dish coating device according to claim 1, wherein a control system (6) is further arranged on the top plate (4), a pressure sensing device (11) is arranged between the telescopic mechanism (10) and the coating device (12), the pressure sensing device (11) is used for sensing the end pressure of the coating device (12), and the control system (6) is used for receiving signals of the pressure sensing device (11) and controlling the telescopic mechanism (10) to stretch.
5. Coating method of an automatic multizone simultaneous coating culture dish device according to any of the claims 1-4, characterized in that:
s1: determining the microorganism type and the number of coating areas, and attaching a drip tube (13) and a corresponding coating device (12);
s2: then the culture dish cover is taken down, and the culture dish is placed in the culture dish groove on the bottom plate;
s3: one end of a high-temperature-resistant hose (14) connected with a peristaltic pump (15) is connected to a bacteria liquid bottle (16), the other end of the high-temperature-resistant hose is connected to a liquid dropping pipe (13), the peristaltic pump (15) is started, and the microbial suspension is extracted until the microbial suspension reaches an experimental set value, and the peristaltic pump (15) is stopped;
s4: after the peristaltic pump (15) is stopped, the telescopic mechanism (10) stretches the coating device (12) to the culture dish, and the motor (9) controls the coating device (12) to perform coating movement through a set coating area and microorganism types;
s5: after the coating movement is finished, the telescopic mechanism (10) contracts the coating device (12) to enable the coating device to recover to the initial position, the telescopic mechanism (10) stops working, and the whole coating work is completed.
6. The method of coating an automatic multizone simultaneous coating culture dish device according to claim 5, wherein the coating device (12), the drip tube (13), the high temperature resistant hose (14) and the bacteria liquid bottle (16) all require high pressure steam sterilization, drying and ultraviolet sterilization before use, all of which are done in an ultra clean bench.
7. The coating method of the device for automatically and synchronously coating the culture dishes in multiple areas according to claim 5, wherein a telescopic mechanism (10) is arranged at one side, close to the bottom plate (1), of the center of the top plate (4), a pressure sensing device (11) is arranged between the telescopic mechanism (10) and the coating device (12), and when the bottom of the coating device (12) touches the culture dishes, the pressure sensing device (11) senses pressure, so that the telescopic mechanism (10) stops stretching.
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