CN115558587A - In-situ culture device for microorganism observation and control method - Google Patents

In-situ culture device for microorganism observation and control method Download PDF

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CN115558587A
CN115558587A CN202211288582.9A CN202211288582A CN115558587A CN 115558587 A CN115558587 A CN 115558587A CN 202211288582 A CN202211288582 A CN 202211288582A CN 115558587 A CN115558587 A CN 115558587A
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culture
temperature
membrane
situ
observation
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葛明锋
董文飞
吴彤
宋一之
梅茜
常智敏
李力
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Suzhou Institute of Biomedical Engineering and Technology of CAS
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Suzhou Institute of Biomedical Engineering and Technology of CAS
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    • C12M23/00—Constructional details, e.g. recesses, hinges
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    • C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
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    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
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    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
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    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/30—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration
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    • C12Q3/00—Condition responsive control processes
    • G—PHYSICS
    • G05—CONTROLLING; REGULATING
    • G05B—CONTROL OR REGULATING SYSTEMS IN GENERAL; FUNCTIONAL ELEMENTS OF SUCH SYSTEMS; MONITORING OR TESTING ARRANGEMENTS FOR SUCH SYSTEMS OR ELEMENTS
    • G05B13/00—Adaptive control systems, i.e. systems automatically adjusting themselves to have a performance which is optimum according to some preassigned criterion
    • G05B13/02—Adaptive control systems, i.e. systems automatically adjusting themselves to have a performance which is optimum according to some preassigned criterion electric
    • G05B13/04—Adaptive control systems, i.e. systems automatically adjusting themselves to have a performance which is optimum according to some preassigned criterion electric involving the use of models or simulators
    • G05B13/042—Adaptive control systems, i.e. systems automatically adjusting themselves to have a performance which is optimum according to some preassigned criterion electric involving the use of models or simulators in which a parameter or coefficient is automatically adjusted to optimise the performance

Abstract

The invention discloses an in-situ culture device for microorganism observation, which belongs to the field of microorganism culture, wherein a gas mixing device is used for mixing gas of a plurality of gas sources, a culture dish comprises a body, a bracket and a culture membrane, the bracket is arranged on the body, the culture membrane is arranged on the bracket and is positioned in a culture area, the body is provided with a liquid inlet and outlet, the liquid inlet and outlet is connected with an external injection pump, the liquid inlet and outlet is also communicated with the culture area, and the temperature of the culture box can be controlled to be uniform through the arrangement of a first heating membrane, a plurality of second heating membranes and a plurality of temperature sensors; through the setting of culture dish business turn over liquid mouth and culture membrane, make culture apparatus can adjust the culture solution under airtight condition, be transparent design through the box part for culture apparatus in situ who can last in real time observes. The invention also relates to a control method of the in-situ culture device for observing the microorganisms.

Description

In-situ culture device for microorganism observation and control method
Technical Field
The invention relates to the field of microorganism culture, in particular to an in-situ culture device for microorganism observation and a control method.
Background
With the continuous development of technology, monitoring of the microbial culture process is also more and more important. For example, due to abuse or misuse of antibiotics, drug-resistant bacteria, multi-drug-resistant bacteria and super bacteria emerge endlessly, and in recent years, researchers find that a very low proportion of individuals in a population of pathogenic bacteria can enter a retention state, which is one of the important mechanisms causing drug resistance. Since the retention phenomenon of bacteria is easily changed by the influence of external environment, and the retention state is a phenotype state of the bacteria, real-time continuous in-situ culture observation is very necessary for the research of the retention bacteria. In addition, the culture conditions of the bacteria difficult to culture, such as the type and concentration of the culture medium and the research on growth factors, are also the key to discover the novel culturable bacteria, and the continuous dynamic in-situ culture and observation of the microorganisms are also required.
The existing in-situ culture devices are all observed aiming at living cells, and have the problems of insufficient temperature uniformity, the culture solution cannot be adjusted under a closed condition, and the requirement of continuous in-situ culture experiments of microorganisms cannot be completely met.
Disclosure of Invention
In order to overcome the defects of the prior art, the invention aims to provide an in-situ culture device for microorganism observation, which can continuously perform in-situ culture observation in real time, has uniform temperature and can adjust a culture solution under a closed condition.
In order to overcome the defects of the prior art, the invention also aims to provide a control method of the in-situ culture device for microorganism observation, which can continuously perform in-situ culture observation in real time, has uniform temperature and can adjust the culture solution under a closed condition.
One of the purposes of the invention is realized by adopting the following technical scheme:
the utility model provides an in situ culture device for microorganism is surveyd, includes incubator, a plurality of air supply and gas mixing device, and is a plurality of the air supply with gas mixing device intercommunication, gas mixing device makes a plurality ofly the gas mixture of air supply, the incubator include the box and install in the inside culture dish of box, the culture dish includes body, support and cultivates the membrane, the support install in the body, cultivate the membrane install in the support is located the cultivation region, the body is equipped with business turn over liquid mouth, business turn over liquid mouth is connected with outside syringe pump, business turn over liquid mouth still with cultivate regional intercommunication, the box part is made for transparent material, the box includes first heating film, a plurality of second heating film and a plurality of temperature sensor, first heating film, a plurality of the second heating film encircles the culture dish, temperature sensor encircles the culture dish.
Further, the box still includes the upper cover, the upper cover include first transparent plate, intercommunication connect and first heating film, first heating film is fixed in first transparent plate, the intercommunication connect install in first transparent plate, the business turn over liquid mouth passes through the intercommunication connects and is connected with outside syringe pump.
Further, the box still includes the lateral wall, the lateral wall includes the main part and is fixed in the main part the second heats the membrane, the quantity of second heating membrane is a plurality of, and is a plurality of the second heats the membrane and sets up respectively in the relative both sides of lateral wall.
Furthermore, the box includes the base, the base includes the second transparent plate and is fixed in a plurality of the second transparent plate temperature sensor, temperature sensor's quantity is five temperature sensor is located respectively the four corners and the center of second transparent plate.
Further, the in-situ culture device for observing the microorganisms further comprises a humidity control device, wherein the humidity control device is positioned between the gas mixing device and the incubator, and the humidity control device is respectively communicated with the gas mixing device and the incubator.
The second purpose of the invention is realized by adopting the following technical scheme:
a control method of the in-situ culture device for observing the microorganisms comprises the following steps:
temperature control: the pelican algorithm is adopted to generate a neural network initial weight matrix, then the BP neural network is combined with the PID algorithm, so that PID parameters can be adjusted in real time, the self-adaption capability of the PID algorithm is improved, the overshoot of the system is reduced, and the high uniformity of the temperature in the micro-culture system is ensured by adopting a distributed temperature control technology combined with the weight matrix;
controlling gas: a sensor in the gas mixing device is adopted to monitor the gas mixing degree, so that the gas in the incubator can be accurately controlled;
humidity control: humidifying the gas by adopting a humidity control device and inputting the humidified gas into an incubator to ensure that the ambient humidity in the incubator is more than 90%;
controlling a culture solution: inside passing through business turn over liquid mouth and the intercommunication connects and is connected with outside accurate syringe pump, and accurate syringe pump realizes that the little upgrading culture solution absorbs and fills the operation, has arranged the permeable nanometer PTFE membrane of one deck culture solution in the culture dish culture region, and the culture is inoculated on the surface of membrane for the in-process culture of changing the culture solution can not flow along with the culture solution.
Further, in the temperature control step, the pelican algorithm specifically comprises: the method comprises two updating stages of approaching prey and water surface flight, wherein in the prey approaching stage, an updating formula is as follows:
Figure BDA0003900407050000021
in the formula: x i,j In order to update the position of the previous individual,
Figure BDA0003900407050000022
for the updated individual position, i represents an individual number, and j represents an individual dimension; rand is a random number, I is a 1 or 2 random integer; p is a radical of formula j Randomly generated individual jth dimension values; f is an objective function value;
after the prey stage is updated, accepting or rejecting the updated individual according to a greedy strategy;
the formula of the water surface flight phase is as follows, and the phase can carry out more full early global search and more accurate later local search:
Figure BDA0003900407050000031
in the formula: r is a random integer of 0 or 2; t is the current iteration number, and T is the total iteration number.
Further, in the temperature control step, a fuzzy control method is adopted to carry out grading quantification on the influence effect of the heating film j on the temperature control point i by the action effect of the heating film and endow a proper weight p by combining with experimental experience ji And the distributed temperature control technology combined with the weight matrix ensures that the temperature of the edge control point is stabilized at the target temperature, and simultaneously compensates the temperature of the middle low-temperature area, thereby improving the temperature uniformity of the system.
Furthermore, the output power P of different heating films is related to the regulating quantity u (T) and the weighting coefficient rho which are obtained by calculating the feedback temperature of each temperature control point through an algorithm
Figure BDA0003900407050000032
Compared with the prior art, the support of the culture dish of the in-situ culture device for observing microorganisms is arranged on the body, the culture membrane is arranged on the support and is positioned in the culture area, the body is provided with the liquid inlet and outlet, the liquid inlet and outlet are connected with an external injection pump and are also communicated with the culture area, the box body is made of transparent materials, the box body comprises a first heating membrane, a plurality of second heating membranes and a plurality of temperature sensors, the first heating membrane and the plurality of second heating membranes surround the culture dish, the temperature sensors surround the culture dish, and the temperature of the culture box can be controlled to be uniform through the arrangement of the first heating membrane, the plurality of second heating membranes and the plurality of temperature sensors; through the setting of culture dish business turn over liquid mouth and culture membrane, make culture apparatus can adjust the culture solution under airtight condition, be transparent design through the box part for culture apparatus can real-time continuous in situ observation.
Drawings
FIG. 1 is a schematic view of the in situ culture apparatus for observing microorganisms according to the present invention;
FIG. 2 is a perspective view of a gas mixing device of the in situ culture apparatus for observing microorganisms of FIG. 1;
FIG. 3 is a perspective view of an incubator of the in situ culture apparatus for observing microorganisms of FIG. 1;
FIG. 4 is a schematic view showing an internal structure of the cultivation box of FIG. 3;
FIG. 5 is a perspective view of the upper lid of the incubator of FIG. 3;
FIG. 6 is a perspective view of the base of the incubator of FIG. 3;
FIG. 7 is a perspective view of the culture dish of the incubator of FIG. 3;
FIG. 8 is a distribution diagram of temperature sensors of the incubator of FIG. 3;
FIG. 9 is a diagram of a single-layer neural network structure.
In the figure: 10. a gas source; 20. a gas mixing device; 21. a housing; 22. a fan electrical interface; 23. an air inlet; 24. a sensor electrical interface; 25. an air outlet; 30. a humidity control device; 40. an incubator; 41. a box body; 411. an upper cover; 4110. an upper frame; 4111. a first transparent plate; 4112. a communication joint; 4113. a first heating film; 412. a side wall; 4120. a main body; 4121. a second heating film; 4122. a cable joint; 4123. a quick connector; 4124. an aviation plug; 413. a base; 4130. a lower frame; 4131. a second transparent plate; 4132. a temperature sensor; 42. a culture dish; 420. a body; 421. a liquid inlet and outlet; 422. a support; 423. and (5) culturing the membrane.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
It will be understood that when an element is referred to as being "secured to" another element, it can be directly on the other element or intervening elements may also be present, secured by intervening elements. When a component is referred to as being "connected" to another component, it can be directly connected to the other component or intervening components may also be present. When a component is referred to as being "disposed on" another component, it can be directly disposed on the other component or intervening components may also be present. The terms "vertical," "horizontal," "left," "right," and the like as used herein are for illustrative purposes only.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.
As shown in FIG. 1, the in situ culture apparatus for observing microorganisms of the present inventionThe device comprises a gas source 10, a gas mixing device 20, a humidity control device 30 and an incubator 40. The number of the gas sources 10 is plural, and the plural gas sources 10 respectively contain CO 2 ，O 2 ，N 2 And the like. The gas mixing device 20 is connected with the plurality of gas sources 10, the gas mixing device 20 mixes the gas input by the plurality of gas sources 10, the humidity control device 30 is communicated with the gas mixing device 20, the humidity control device 30 carries out ultrasonic atomization and humidification on the gas input by the gas mixing device 20, and the humidified gas enters the incubator 40.
Referring to fig. 2, the gas mixing device 20 includes a housing 21 and a fan installed inside the housing 21, and the fan rotates to mix the gas inside the housing 21. The casing 21 is provided with a fan electrical interface 22, an air inlet 23, a sensor electrical interface 24 and an air outlet 25, and the fan electrical interface 22 is connected with a fan to provide power for the fan. The gas inlets 23 are connected to the plurality of gas sources 10, so that the gas from the gas sources 10 enters the gas mixing device 20. The sensor electrical interface 24 is connected to the sensor within the housing 21. The sensor is used for detecting the gas concentration and can monitor the gas condition in real time.
Humidity control device 30 is inside to be equipped with ultrasonic atomization humidifier and humidity transducer, and gaseous entering incubator 40 behind the intensive mixing in humidity control device 30 cavity through ultrasonic atomization humidifier, humidity is stopped by the monitoring of inside humidity transducer and according to real-time humidity value feedback control ultrasonic humidifier work start-stop in incubator 40 for incubator 40 inside humidity is invariable.
With continued reference to fig. 3-7, the incubator 40 includes a housing 41 and a culture dish 42 mounted to the housing 41.
The case 41 includes an upper cover 411, a sidewall 412, and a base 413.
The upper cover 411 includes an upper frame 4110, a first transparent plate 4111, a communication joint 4112, and a first heating film 4113. First transparent board 4111 is fixed to upper frame 4110, and first transparent board 4111 is made of acrylic board. First transparent panel 4111 facilitates an operator's observation of the microorganism growth process. And the communication joint 4112 is a luer joint and is used for inputting a culture solution. Luer connectors are used as culture fluid input and output interfaces, and a precision injection pump outside the incubator 40 can be connected to the luer connectors of the internal culture dish 42 through pipelines to realize micro-upgrading culture fluid suction and perfusion operations. The first heating film 4113 is attached to the first transparent plate 4111, and the first heating film 4113 is a transparent ITO heating film.
The side wall 412 includes a body 4120, a second heating membrane 4121, a cable connector 4122, a quick-connect connector 4123, and an aircraft plug 4124. The main body 4120 is made of an aluminum alloy material with good thermal conductivity, and heat insulation cotton with low thermal conductivity is placed on the outer wall. The number of the second heating films 4121 is plural, and the plural second heating films 4121 are fixed to opposite sides of the sidewall 412. Specifically, the number of the second heating films 4121 is four, and two second heating films 4121 are fixed to two opposite walls, respectively. The second heating film 4121 is a PI heating film. The cable connector 4122 is used for sensor cable entry. Quick connector 4123 is used for gas ingress and egress. The aircraft plug 4124 is used for power cable entry.
The base 413 includes a lower bezel 4130, a second transparent plate 4131, and a temperature sensor 4132. The second transparent plate 4131 is made of glass, and the second transparent plate 4131 is fixed to the lower frame 4130. The number of the temperature sensors 4132 is plural, and the plural temperature sensors 4132 are fixed to the second transparent plate 4131. Specifically, in the present embodiment, the number of the temperature sensors 4132 is five, four temperature sensors 4132 are located at four corners of the second transparent plate 4131, and another temperature sensor 4132 is located in the middle of the second transparent plate 4131. To ensure the temperature uniformity of the culture core region 50mm by 5mm, a control arrangement of 5 temperature monitoring points is shown in FIG. 8.
The upper cover 411, the side wall 412 and the base 413 are fixed to each other to form the cultivation box 40.
The culture dish 42 includes a body 420, a support 422, and a culture membrane 423. The body 420 is internally provided with a culture area, the body 420 is externally provided with a liquid inlet and outlet 421, and the liquid inlet and outlet 421 is communicated with the culture area through a micro-channel. The holder 422 is attached to the body 420, and the culture membrane 423 is attached to the holder 422. The culture membrane 423 is a nano-scale PTFE membrane. The culture membrane 423 is located on the culture region. The culture is inoculated on the surface of the membrane, so that the culture does not flow out along with the culture solution in the process of replacing the culture solution, and the real-time change of the components of the culture solution under the sealed condition of the incubator 40 is realized.
The invention also relates to a control method of the in-situ culture device for observing the microorganisms, which comprises the following steps:
temperature control: generating a neural network initial weight matrix by using a pelican algorithm, then combining a BP neural network with a PID algorithm, so that PID parameters can be adjusted in real time, the self-adaptive capacity of the PID algorithm is improved, the overshoot of the system is reduced, and the high uniformity of the temperature in the micro-culture system is ensured by using a distributed temperature control technology combined with the weight matrix;
controlling gas: a sensor in the gas mixing device 20 is adopted to monitor the gas mixing degree, so that the gas in the incubator 40 can be accurately controlled;
humidity control: humidifying the gas by adopting a humidity control device 30, and inputting the humidified gas into an incubator 40 to ensure that the ambient humidity in the incubator 40 is more than 90%;
controlling the culture solution: the inside business turn over liquid mouth 421 and the intercommunication connects 4112 of passing through of culture dish 42 is connected with outside accurate syringe pump, and accurate syringe pump realizes that the little upgrading culture solution absorbs and fills the operation, has arranged the permeable nanometer PTFE membrane of one deck culture solution in culture dish 42's the culture region, and the culture is inoculated on the surface of membrane for the in-process culture of changing the culture solution can not flow out along with the culture solution.
Specifically, the temperature control step specifically comprises:
the temperature control system of the incubator 40 is a first-order inertia delay system which is easy to have the problem of control overshoot, a BP neural network is introduced to be combined with a PID algorithm, so that PID parameters can be adjusted in real time, the self-adaptive capacity of the PID algorithm is improved, and the overshoot of the system is reduced.
Because the initial weight of the BP neural network is randomly generated, the final parameter optimization result is poor in repeatability, and a better PID parameter value cannot be accurately updated each time, the pelican algorithm is introduced to generate the initial weight matrix of the neural network, the repeatability of the algorithm is improved, compared with the traditional group optimization algorithm, the pelican algorithm has the characteristics of simple structure and strong local and global optimizing capability, the algorithm mainly comprises two main updating stages of approaching a prey and flying on the water, wherein in the stage of approaching the prey, after the updating according to the following formula, updated individuals are chosen according to a greedy strategy:
Figure BDA0003900407050000061
in the formula: x i,j In order to update the position of the previous individual,
Figure BDA0003900407050000062
for the updated individual position, i represents the individual number, and j represents the individual dimension; rand is a random number, and I is a random integer of 1 or 2; p is a radical of j Randomly generated individual jth dimension values; f is the objective function value.
The formula of the water surface flight phase is as follows, and the phase can carry out more full early global search and more accurate later local search:
Figure BDA0003900407050000063
in the formula: r is a random integer of 0 or 2; t is the current iteration number, and T is the total iteration number.
On the basis of the algorithm, the distributed temperature control technology combined with the weight matrix is adopted to ensure the high uniformity of the temperature in the micro culture system, and CO outside the culture box 40 is adopted 2 ，O 2 ，N 2 When the gas is uniformly mixed and monitored, the gas in the incubator 40 is accurately controlled, and the gas is humidified by an ultrasonic atomization humidifierThen, the temperature of the micro-incubator 40 is stabilized to be more than 90% RH by feeding the micro-incubator 40. And finally, a culture solution control module is added in the incubator 40, and the culture solution in the culture dish 42 is adjusted in real time under a closed condition through a micro-upgrading precision sample adding channel.
Combining the characteristic that the temperature of a core control area of the system is high near the edge of the heating element and the temperature of a middle area of the heating element is low, and the relation of coupling heat transfer among 5 heating modules, the system adopts a fuzzy control method, carries out grading quantification on the influence effect of a heating film j on a temperature control point i by the action effect of the heating film, and endows a proper weight rho by combining experimental experience ji The grading quantization result is shown in table 1, and the distributed temperature control technology combined with the weight matrix ensures that the temperature of the edge control point is stabilized at the target temperature, compensates the temperature of the middle low-temperature area, and improves the temperature uniformity of the system.
TABLE 1 hierarchical quantization table
Figure BDA0003900407050000071
Finally, the output power P of different heating films is in relation with the regulating quantity u (T) and the weighting coefficient rho which are obtained by calculating the feedback temperature of each temperature control point through an algorithm.
Figure BDA0003900407050000072
The PID algorithm formula is specifically as follows:
u(k)＝K p [e(k)-e(k-1)]+K i e(k)+K d [e(k)-2e(k-1)+e(k-2)] (4)
in the formula: k is p Is a proportionality coefficient, K i 、K d And e (k) is the difference between a set value and the current value of the controlled parameter, u (k) is the actual control parameter of the actuating element, e (k-1) is the difference between the previous controlled parameter and the set value, and e (k-2) is the difference between the previous controlled parameter and the set value.
The in-situ culture device for observing the microorganisms has small volume and good portability. The system is provided with a culture solution control module, can realize micro-upgrading precise sample adding, and can complete the regulation of a microorganism culture solution without changing the culture condition in the in-situ culture process. The system adopts an incremental PID algorithm to realize high-precision control of the temperature of an in-situ culture system for microorganism observation, and the temperature control precision reaches +/-0.10 ℃. The temperature uniformity in the micro in-situ culture system is guaranteed to be +/-0.20 ℃ by adopting a distributed temperature control technology introducing a weight matrix.
The above examples only show some embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for those skilled in the art, variations and modifications can be made without departing from the spirit of the invention, and all equivalent modifications and changes can be made to the above embodiments according to the essential technology of the invention, which falls into the protection scope of the invention.

Claims (9)

1. The utility model provides an in situ culture device for microorganism is surveyd, includes the incubator, its characterized in that: the in-situ culture device for microorganism observation further comprises a plurality of air sources and a gas mixing device, the air sources are communicated with the gas mixing device, the gas mixing device enables the air sources to be multiple, the incubator comprises a box body and a culture dish arranged inside the box body, the culture dish comprises a body, a support and a culture membrane, a support is arranged on the body, the culture membrane is arranged on the support and located in a culture area, the body is provided with a liquid inlet and outlet, the liquid inlet and outlet is connected with an external injection pump, the liquid inlet and outlet is further communicated with the culture area, the box body is made of a transparent material, the box body comprises a first heating membrane, a plurality of second heating membranes and a plurality of temperature sensors, the first heating membrane, the plurality of second heating membranes surround the culture dish, and the temperature sensors surround the culture dish.
2. The in situ culture apparatus for microbiological observation according to claim 1, wherein: the box still includes the upper cover, the upper cover include first transparent plate, intercommunication connect and first heating film, first heating film is fixed in first transparent plate, the intercommunication connect install in first transparent plate, the business turn over liquid mouth passes through the intercommunication connects and is connected with outside syringe pump.
3. The in situ culture apparatus for microbiological observation of claim 1, wherein: the box still includes the lateral wall, the lateral wall includes the main part and is fixed in the main part the second heats the membrane, the quantity of second heating membrane is a plurality of, and is a plurality of the second heat the membrane set up respectively in the relative both sides of lateral wall.
4. The in situ culture apparatus for microbiological observation of claim 1, wherein: the box includes the base, the base includes the second transparent plate and is fixed in a plurality of the second transparent plate temperature sensor, temperature sensor's quantity is five temperature sensor is located respectively the four corners and the center of second transparent plate.
5. The in situ culture apparatus for microbiological observation of claim 1, wherein: the in-situ culture device for microorganism observation further comprises a humidity control device, wherein the humidity control device is positioned between the gas mixing device and the incubator, and the humidity control device is respectively communicated with the gas mixing device and the incubator.
6. A control method of the in situ culture apparatus for observation of microorganisms according to any one of claims 1 to 5, comprising the steps of:
temperature control: generating a neural network initial weight matrix by using a pelican algorithm, then combining a BP neural network with a PID algorithm, so that PID parameters can be adjusted in real time, the self-adaptive capacity of the PID algorithm is improved, the overshoot of the system is reduced, and the high uniformity of the temperature in the micro-culture system is ensured by using a distributed temperature control technology combined with the weight matrix;
controlling gas: a sensor in the gas mixing device is adopted to monitor the gas mixing degree, so that the gas in the incubator can be accurately controlled;
humidity control: humidifying the gas by adopting a humidity control device and inputting the humidified gas into an incubator to ensure that the ambient humidity in the incubator is more than 90%;
controlling a culture solution: inside passing through business turn over liquid mouth and the intercommunication connects and is connected with outside accurate syringe pump, and accurate syringe pump realizes that the micro-upgrade culture solution absorbs and fills the operation, has arranged the permeable nanometer PTFE membrane of one deck culture solution in the culture dish culture region, and the culture is inoculated on the surface of membrane for the in-process culture of changing the culture solution can not flow along with the culture solution.
7. The control method of the in-situ culture apparatus for observation of microorganisms according to claim 6, wherein: in the temperature control step, the pelican algorithm specifically comprises: the method comprises two updating stages of approaching prey and water surface flight, wherein in the prey approaching stage, an updating formula is as follows:
Figure FDA0003900407040000021
in the formula: x i,j In order to update the position of the previous individual,
Figure FDA0003900407040000022
for the updated individual position, i represents an individual number, and j represents an individual dimension; rand is a random number, and I is a random integer of 1 or 2; p is a radical of formula j Randomly generated individual jth dimension values; f is a target function value;
after the prey stage is updated, accepting or rejecting the updated individual according to a greedy strategy;
the formula of the water surface flight phase is as follows, and the phase can carry out more full early global search and more accurate later local search:
Figure FDA0003900407040000023
in the formula: r is a random integer of 0 or 2; t is the current iteration number, and T is the total iteration number.
8. The control method of the in-situ culture apparatus for observation of microorganisms according to claim 7, wherein: in the temperature control step, a fuzzy control method is adopted to carry out grading quantification on the influence effect of the heating film j on the temperature control point i by the action effect of the heating film and endow a proper weight p by combining with experimental experience ji And the distributed temperature control technology combined with the weight matrix ensures that the temperature of the edge control point is stabilized at the target temperature, and simultaneously, the temperature of the middle low-temperature area is compensated, so that the temperature uniformity of the system is improved.
9. The control method of the in-situ culture apparatus for observation of microorganisms according to claim 8, wherein: the output power P of different heating films is related to the regulating quantity u (T) and the weighting coefficient rho which are obtained by the algorithm calculation according to the feedback temperature of each temperature control point
Figure FDA0003900407040000024
CN202211288582.9A 2022-10-20 2022-10-20 In-situ culture device for microorganism observation and control method Pending CN115558587A (en)

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