Microscope live culture environment control system and method

Abstract

The application discloses a microscope living body culture environment control system and method, relates to the monitoring control technical field, can be applied to the microscopic observation demand in the field of cell biology and medicine, and mainly solves the problem of heating and airflow disturbance interference micro-imaging of the existing microscope living body sample culture device. Mainly includes embedded controller, monitoring component, temperature control component and gas control component; wherein, the embedded controller determines real-time temperature monitoring data and real-time concentration of target gas based on real-time temperature sensing signal and gas concentration sensing signal; calculates temperature deviation and gas concentration deviation; when the temperature deviation and / or gas concentration deviation does not satisfy the corresponding target parameter deviation condition, determines the target control component and the control signal of the target control component according to the deviation that does not satisfy the target parameter deviation condition; the control signal is sent to the target control component. The application is mainly used for controlling the environmental parameters of living cells under microscope observation.

Description

Technical Field

[0001] This invention relates to the field of monitoring and control technology, and can be applied to the microscopic observation needs in cell biology and medicine. In particular, it relates to a microscopic in vivo culture environment control system and method. Background Technology

[0002] In cutting-edge fields such as cell biology, drug development, and regenerative medicine, maintaining a stable and precise in vitro culture environment for live cells is the cornerstone of experimental success. Core environmental parameters within the cell culture chamber, especially temperature and the concentration of specific gases (such as carbon dioxide), directly determine the metabolic activity, proliferation rate, gene expression, and even the survival status of cells. Temperature is a key physical factor for enzymatic reactions and cell membrane fluidity, while carbon dioxide concentration regulates a series of intracellular biochemical processes by affecting the pH of the culture medium. Therefore, real-time monitoring and dynamic adjustment of temperature and gas concentration within a live cell workstation are crucial for simulating the in vivo physiological microenvironment, ensuring experimental reproducibility and data reliability, and are of paramount importance for obtaining truly effective biological observation results.

[0003] Existing live-cell environment control systems primarily use proportional gas for concentration control. However, this method requires continuous gas injection, which can cause gas flow interference with the imaging system. Furthermore, existing environmental control systems suffer from low temperature control accuracy, long feedback cycles, and are prone to temperature fluctuations, leading to thermal expansion and contraction in the imaging system. As a highly sensitive system, the microscope is susceptible to interference from temperature or airflow disturbances in its microscopic imaging quality. In summary, existing live-cell sample culture devices for microscopes suffer from problems related to thermal and airflow disturbances interfering with microscopic imaging. Summary of the Invention

[0004] In view of this, the present invention provides a control system and method for a live culture environment for microscopes, the main purpose of which is to solve the problem of interference with microscopic imaging caused by heat and airflow disturbances in existing live sample culture devices for microscopes.

[0005] According to one aspect of the present invention, a microscopic in vivo culture environment control system is provided, comprising: An embedded controller, a monitoring component, a temperature control component, and a gas control component; the embedded controller communicates with the monitoring component via an interface and controls the temperature control component and the gas control component. The monitoring component is used to generate and send real-time temperature sensing signals of the target workstation and gas concentration sensing signals of the target gas in the cell culture chamber to the embedded controller in real time. The embedded controller is configured to determine real-time temperature monitoring data and the real-time concentration of the target gas based on the real-time temperature sensing signal and the gas concentration sensing signal; calculate the temperature deviation between the real-time temperature monitoring data and the target temperature, and calculate the gas concentration deviation between the real-time concentration and the target gas concentration; when the temperature deviation and / or the gas concentration deviation do not meet the corresponding target parameter deviation conditions, determine the target control component and the control signal of the target control component based on the deviation that does not meet the target parameter deviation conditions; and send the control signal to the target control component, wherein the target control component includes a temperature control component and / or a gas control component. The temperature control component is used to perform temperature adjustment according to the control signal; The gas control component is used to perform gas concentration adjustment based on the control signal.

[0006] Furthermore, the gas monitoring component includes a temperature sensor, a temperature sensing processing circuit, and a non-dispersive gas sensor. The temperature sensor is used to generate and output resistance data based on the temperature change of the measurement area; The temperature sensing processing circuit is used to convert the resistance data output by the temperature sensor into analog voltage data. The analog-to-digital converter circuit is used to convert the analog voltage data into real-time temperature monitoring data; The non-dispersive gas sensor is used to generate a gas concentration sensing signal based on the target gas concentration within the measurement area.

[0007] Furthermore, the embedded controller is specifically used to determine the target control component as a gas control component and determine the control signal of the gas control component when the gas concentration deviation does not meet the corresponding target parameter deviation condition. The gas control component includes a normally closed solenoid valve for controlling the target gas input and a normally closed air pump for controlling the air input. The control signal determination process of the gas control component includes: The expected adjustment amount of the target gas is calculated based on the gas concentration deviation. Based on the expected adjustment amount, under the constraint of total gas balance in the culture chamber, a first pulse width modulation signal for controlling the normally closed gas pump and a second pulse width modulation signal for controlling the normally closed solenoid valve are generated collaboratively.

[0008] Furthermore, the embedded controller is specifically used to determine the target control component as a temperature control component when the temperature deviation does not meet the corresponding target parameter deviation condition, and to determine the control signal of the temperature control component, wherein the temperature control component includes a gas temperature control component and an ambient temperature control component. The process of determining the control signal for any temperature control component includes: When the temperature deviation is less than a preset threshold, the pulse width modulation signal controls the temperature control component to perform equivalent power continuously adjustable heating with a corresponding duty cycle, thereby adaptively achieving temperature stability. When the temperature deviation exceeds a preset threshold, the pulse width modulation signal is in an invalid state or the temperature control component is stopped working.

[0009] Furthermore, the gas temperature control component is a water bottle heating sleeve, which surrounds the mixed gas water bottle; the ambient temperature control component is a ceramic heating element, which is arranged in the non-observation area of ​​the cell culture chamber.

[0010] Furthermore, the target temperature includes the target gas temperature, the gas control component includes a gas-mixing bottle, and the embedded control system is specifically used to acquire the gas intake of the gas-mixing bottle and calculate the gas intake deviation value between the gas intake and a preset gas intake threshold. The gas temperature compensation value is calculated based on the intake volume deviation value. The target gas temperature is then positively corrected using the gas temperature compensation value, so that the corrected target gas temperature is used as the target gas temperature in the subsequent calculation of the gas temperature deviation.

[0011] Furthermore, the system also includes a dual MOS drive circuit; The dual MOS driving circuit is used to receive the microsecond-level high-frequency control signal sent by the embedded controller, amplify the control signal to obtain a driving signal, and send the driving signal to the corresponding target control component to drive the target control component to perform gas concentration and / or temperature regulation.

[0012] Furthermore, the system also includes interactive components; The embedded controller is also used to output the real-time temperature monitoring data of the target workstation and the real-time concentration of the target gas in the cell culture chamber to the interactive component; The interactive component is used to display the real-time temperature monitoring data and the real-time concentration of the target gas; The interactive component is also used to acquire and forward to the embedded controller the target gas temperature, target ambient temperature, target gas concentration, and target parameter deviation conditions input by the user.

[0013] According to another aspect of the present invention, a method for controlling the in vivo culture environment for microscopes is provided, comprising: The monitoring component generates and sends real-time temperature sensing signals of the target workstation and gas concentration sensing signals of the target gas in the cell culture chamber to the embedded controller in real time. The embedded controller determines real-time temperature monitoring data and the real-time concentration of the target gas based on the real-time temperature sensing signal and the gas concentration sensing signal; calculates the temperature deviation between the real-time temperature monitoring data and the target temperature, and calculates the gas concentration deviation between the real-time concentration and the target gas concentration; when the temperature deviation and / or the gas concentration deviation do not meet the corresponding target parameter deviation conditions, the controller determines the target control component and the control signal of the target control component based on the deviation that does not meet the target parameter deviation conditions; and sends the control signal to the target control component, wherein the target control component includes a temperature control component and / or a gas control component. Temperature regulation is performed by the temperature control component according to the control signal; The gas concentration is adjusted according to the control signal via the gas control component.

[0014] Furthermore, the real-time generation and transmission of the real-time temperature sensing signal of the target workstation and the gas concentration sensing signal of the target gas in the cell culture chamber to the embedded controller includes: Resistance data is generated and output based on temperature changes in the measurement area using a temperature sensor. The resistance data output by the temperature sensor is converted into analog voltage data using a Wheatstone bridge and a differential operational amplifier circuit. The analog voltage data is converted into real-time temperature monitoring data through an analog-to-digital converter circuit. A gas concentration sensing signal is generated based on the target gas concentration within the measurement area using a non-dispersive gas sensor.

[0015] Furthermore, when the gas concentration deviation does not meet the corresponding target parameter deviation condition, the target control component is determined to be a gas control component, wherein the gas control component includes a normally closed solenoid valve for controlling the target gas input and a normally closed air pump for controlling the air input. The control signal determination process of the gas control component includes: The expected adjustment amount of the target gas is calculated based on the gas concentration deviation. Based on the expected adjustment amount, under the constraint of total gas balance in the culture chamber, a first pulse width modulation signal for controlling the normally closed gas pump and a second pulse width modulation signal for controlling the normally closed solenoid valve are generated collaboratively.

[0016] Furthermore, when the temperature deviation does not meet the corresponding target parameter deviation condition, the target control component is determined to be a temperature control component, wherein the temperature control component includes a gas temperature control component and an ambient temperature control component. The process of determining the control signal for any temperature control component includes: When the temperature deviation is less than a preset threshold, the pulse width modulation signal controls the temperature control component to perform equivalent power continuously adjustable heating with a corresponding duty cycle, thereby adaptively achieving temperature stability. When the temperature deviation exceeds a preset threshold, the pulse width modulation signal is in an invalid state or the temperature control component is stopped working.

[0017] Furthermore, the gas temperature control component is a water bottle heating sleeve, which surrounds the mixed gas water bottle; the ambient temperature control component is a ceramic heating element, which is arranged in the non-observation area of ​​the cell culture chamber.

[0018] Furthermore, the target temperature includes the target gas temperature, the gas control component includes a mixed-gas bottle, and the method further includes: Obtain the air intake volume of the mixed-gas bottle and calculate the air intake volume deviation value between the air intake volume and the preset air intake volume threshold; The gas temperature compensation value is calculated based on the intake volume deviation value. The target gas temperature is then positively corrected using the gas temperature compensation value, so that the corrected target gas temperature is used as the target gas temperature in the subsequent calculation of the gas temperature deviation.

[0019] Further, the control signal is sent to the target control component, wherein the target control component includes a temperature control component and / or a gas control component, to drive the target control component to perform gas concentration and / or temperature regulation, including: The control signal is sent to a dual MOS drive circuit to amplify the control signal and obtain a drive signal. This drive signal is then sent to the corresponding target control component to drive the target control component to perform gas concentration and / or temperature regulation.

[0020] Furthermore, the method also includes: The real-time temperature monitoring data of the target workstation and the real-time concentration of the target gas in the cell culture chamber are output to the interactive component to display the real-time temperature monitoring data and the real-time concentration of the target gas. The interactive component acquires and sends the target gas temperature, target ambient temperature, target gas concentration, and target parameter deviation conditions input by the user to the embedded controller.

[0021] According to another aspect of the present invention, a workstation is provided that performs environmental parameter control operations on the cell culture chamber within the workstation based on the above-described microscopic in vivo culture environment control system.

[0022] By employing the above-described technical solutions, the technical solutions provided by the embodiments of the present invention have at least the following advantages: This invention provides a control system and method for a microscopic in vivo culture environment. In this embodiment, a monitoring component generates and sends real-time temperature sensing signals from the target workstation and gas concentration sensing signals from the target gas within the cell culture chamber to an embedded controller. The embedded controller determines real-time temperature monitoring data and the real-time concentration of the target gas based on the real-time temperature and gas concentration sensing signals. It calculates the temperature deviation between the real-time temperature monitoring data and the target temperature, and the gas concentration deviation between the real-time concentration and the target gas concentration. When the temperature deviation and / or gas concentration deviation do not meet the corresponding target parameter deviation conditions, a target control component and its control signal are determined based on the deviation amount that does not meet the target parameter deviation conditions. The control signal is sent to the target control component, which includes a temperature control component and / or a gas control component. The temperature control component performs temperature adjustment based on the control signal, and the gas control component performs gas concentration adjustment based on the control signal. By controlling gas injection based on high-frequency gas detection and real-time feedback, intermittent gas supply is achieved, making gas injection more stable and reducing gas supply vibration and gas consumption. Through microsecond-level high-frequency feedback temperature control, the temperature fluctuation of the imaging system is effectively reduced, thereby reducing the impact of gas fluctuation and temperature fluctuation on microscopic imaging. This reduces the interference of heat and airflow disturbance on microscopic imaging and ensures the quality of microscopic imaging.

[0023] The above description is merely an overview of the technical solution of the present invention. In order to better understand the technical means of the present invention, it can be implemented according to the contents of the specification. Furthermore, in order to make the above and other objects, features and advantages of the present invention more apparent and understandable, specific embodiments of the present invention are described below. Attached Figure Description

[0024] Various other advantages and benefits will become apparent to those skilled in the art upon reading the following detailed description of preferred embodiments. The accompanying drawings are for illustrative purposes only and are not intended to limit the invention. Furthermore, the same reference numerals denote the same parts throughout the drawings. In the drawings: Figure 1 This diagram illustrates a block diagram of a microscopic in vivo culture environment control system provided in an embodiment of the present invention. Figure 2A flowchart of a method for controlling the in vivo culture environment for microscopes, provided by an embodiment of the present invention, is shown. Detailed Implementation

[0025] Exemplary embodiments of the present disclosure will now be described in more detail with reference to the accompanying drawings. While exemplary embodiments of the present disclosure are shown in the drawings, it should be understood that the present disclosure may be implemented in various forms and should not be limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the disclosure to those skilled in the art.

[0026] To address the problem of thermal and airflow disturbances interfering with microscopic imaging in existing microscopic in vivo sample culture devices, this invention provides a microscopic in vivo culture environment control system, such as... Figure 1 As shown, the system includes: an embedded controller 100, a monitoring component 200, a temperature control component 300, and a gas control component 400; the embedded controller 100 communicates with the monitoring component 200, the temperature control component 300, and the gas control component 400 via serial ports through interfaces.

[0027] The monitoring component 200 generates and sends real-time temperature sensing signals from the target workstation and gas concentration sensing signals from the target gas in the cell culture chamber to the embedded controller 100 in real time. Based on the received real-time temperature and gas concentration sensing signals, the embedded controller determines the real-time temperature monitoring data and the real-time concentration of the target gas; calculates the temperature deviation between the real-time temperature monitoring data and the target temperature, and calculates the gas concentration deviation between the real-time concentration and the target gas concentration; when the temperature deviation and / or gas concentration deviation do not meet the corresponding target parameter deviation conditions, it determines the target control component and its control signal based on the deviation amount that does not meet the target parameter deviation conditions; and sends the control signal to the target control component. The target control component includes a temperature control component 300 and / or a gas control component 400. When the target control component is the temperature control component 300, it performs temperature regulation according to the control signal; when the target control component is the gas control component 400, it performs gas concentration regulation according to the control signal.

[0028] In this embodiment of the invention, the core of the system lies in achieving precise and automated maintenance of the temperature and specific gas (such as carbon dioxide and oxygen) concentrations within the culture chamber through real-time sensing, closed-loop control, and collaborative execution. The culture chamber is a microscopic in vivo sample culture device, and this system is particularly suitable for scenarios requiring high-precision and high-stability culture conditions, such as embryo culture, stem cell research, or biopharmaceutical processes. The system mainly comprises four functional modules: a monitoring component 200, an embedded controller 100, a temperature control component 300, and a gas control component 400. These modules are interconnected via serial interfaces or data buses to form a complete closed-loop control circuit.

[0029] The monitoring component 200 includes at least one temperature sensor and a target gas concentration sensor (e.g., an infrared CO2 sensor or an electrochemical oxygen sensor). These sensors are strategically positioned at key locations within the target workstation (e.g., inside an incubator) to ensure the representativeness of the measurements. This component transmits the sensed signals (i.e., real-time temperature and gas concentration signals) to the embedded controller 100, which serves as the basis for dynamic adjustments to the system. The embedded controller 100 is typically built on a microcontroller unit (MCU) or microprocessor (MPU) and incorporates or externally connects the necessary signal conditioning circuitry, analog-to-digital converters (ADCs), and communication interfaces.

[0030] The embedded controller 100 receives raw sensing signals from the monitoring component 200 and processes them through filtering, calibration, and unit conversion to convert them into real-time temperature monitoring data and real-time target gas concentration data with clear physical meaning. The embedded controller 100 internally stores or receives pre-configured target temperature and target gas concentration setpoints. It compares the real-time data with the corresponding target values ​​to calculate two key deviations: temperature deviation (the difference between real-time and target temperatures) and gas concentration deviation (the difference between real-time and target concentrations). Target parameter deviation conditions can be that the absolute value of the temperature deviation is less than 0.1°C and the absolute value of the gas concentration deviation is less than 0.1%. The controller determines whether the calculated deviations meet the above conditions. Once it is determined that the temperature deviation and / or gas concentration deviation do not meet the corresponding deviation conditions, the embedded controller immediately initiates a control decision. That is, based on the deviations that do not meet the conditions, it determines the target control component that needs to be activated. This target control component may be the temperature control component 300, the gas control component 400, or a combination of both when both deviations exceed the limit. Next, the embedded controller 100 calculates a control signal based on a control algorithm, such as a proportional-integral-derivative algorithm or a fuzzy control algorithm, and uses this control signal to drive the actuator. Finally, the embedded controller 100 sends the generated control signal to the determined target control component through the corresponding output interface.

[0031] Temperature control component 300 may include a heating element (such as a resistance wire) that, upon receiving a control signal from embedded controller 100, performs physical adjustments based on the specific content of the signal. For example, if the signal indicates that heating is required, the drive circuit may increase the supply current or duty cycle of the heating element. Gas control component 400 comprises a gas supply source (such as a high-pressure gas cylinder), a precision regulating valve (such as a solenoid proportional valve), and a gas mixing device (such as a gas-water mixing bottle).

[0032] It should be noted that the system, through the collaborative work of the aforementioned modules, forms a highly automated closed loop. The combined use of the embedded controller and high-precision sensors enables high-frequency feedback and intermittent adjustment of temperature and gas, avoiding fluctuations caused by continuous adjustment, thereby reducing the interference of gas and temperature fluctuations on microscope microscopic imaging. The monitoring component 200 continuously provides feedback, the embedded controller 100 continuously performs judgments and calculations, and the control component executes precise correction operations. This continuous cycle dynamically offsets fluctuations caused by environmental disturbances and the culture process itself, stabilizing the temperature and gas concentration in the cell culture chamber near the required target parameters, greatly improving the effectiveness, stability, and reliability of culture condition control.

[0033] In one embodiment of the present invention, for further explanation and limitation, the gas monitoring component includes a temperature sensor, a temperature sensing processing circuit, an analog-to-digital conversion circuit, and a non-dispersive gas sensor.

[0034] The system comprises several components: a temperature sensor that generates and outputs resistance data based on temperature changes in the measurement area; a temperature sensing processing unit that converts the resistance data into analog voltage data; an analog-to-digital converter that converts the analog voltage data into a real-time temperature sensing signal; and a non-dispersive gas sensor that generates a gas concentration sensing signal based on the target gas concentration within the measurement area.

[0035] In this embodiment of the invention, the temperature signal is realized through a temperature sensor (such as a PT100 or PT1000 platinum resistance thermometer). This sensor is placed in a key measurement area within the incubation chamber, and its resistance value changes linearly and predictably with the ambient temperature. The temperature sensor itself generates and outputs resistance data based on the temperature change of the measurement area. Embedded controllers typically process digital or voltage signals. Therefore, the system includes a temperature sensing processing circuit, which can be built based on a Wheatstone bridge and a differential operational amplifier circuit. The Wheatstone bridge converts the minute resistance change of the resistance thermometer into an unbalanced voltage difference. This differential voltage is then fed into the differential operational amplifier circuit for precise amplification, resulting in an analog voltage data with an amplitude suitable for subsequent processing, thus completing the conversion from resistance to voltage. Finally, an analog-to-digital converter (ADC) is responsible for converting the amplified analog voltage data into a digital real-time temperature sensing signal for direct reading and processing by the embedded controller.

[0036] For measuring the concentration of a target gas (such as carbon dioxide), a non-dispersive gas sensor (such as the JW01) is used. This sensor operates based on the characteristic absorption principle of infrared spectra for specific gases, offering advantages such as high selectivity, high accuracy, and long lifespan. This sensor directly generates a sensing signal based on the concentration of the target gas within the measurement area. The matching JW002 chip converts the sensor signal into a serial data stream, i.e., a gas concentration sensing signal, which is then extracted from the serial data stream by an embedded system to calculate the real-time gas concentration data of the target gas.

[0037] Through the aforementioned discrete sensing path design, the monitoring components can independently and with high fidelity capture changes in the two key physical quantities, temperature and gas concentration, laying a solid data foundation for the stability and precise control of the entire closed-loop control system. After receiving these digitized temperature and gas concentration sensing signals, the embedded controller can perform deviation calculations and control decisions.

[0038] In one embodiment of the present invention, for further explanation and limitation, the embedded controller is specifically used to determine the target control component as a gas control component and determine the control signal of the gas control component when the gas concentration deviation does not meet the corresponding target parameter deviation condition.

[0039] The gas control component includes a normally closed solenoid valve for controlling the input amount of the target gas and a normally closed air pump for controlling the input amount of air. The control signal determination process of the gas control component includes: calculating the expected adjustment amount of the target gas based on the gas concentration deviation; and, based on the expected adjustment amount and under the constraint of total gas balance in the culture chamber, collaboratively generating a first pulse width modulation signal for controlling the normally closed air pump and a second pulse width modulation signal for controlling the normally closed solenoid valve.

[0040] In this embodiment of the invention, the gas control component employs a bidirectional adjustable mixed gas supply scheme. A normally closed solenoid valve is connected to the target gas, such as a high-purity carbon dioxide source. It remains closed when not energized to ensure gas safety; upon receiving an opening signal, it injects the target gas into the culture chamber as needed. A normally closed air pump is connected to a filtered ambient air supply line to inject clean air into the culture chamber, primarily serving to dilute the target gas.

[0041] The process of generating control signals by the embedded controller is not a simple switch operation, but rather a coordinated regulation strategy based on flow calculation. The control signal determination process specifically includes: calculating the expected adjustment amount. Based on the magnitude and direction (positive or negative) of the gas concentration deviation, combined with parameters such as the known volume of the culture chamber and the gas diffusion model, the controller calculates the expected adjustment amount of the target gas required to correct the deviation (usually in standard volume or moles). This then generates a coordinated pulse width modulation (PWM) signal. To quickly and smoothly adjust the gas concentration to the target value, while avoiding drastic pressure fluctuations within the culture chamber, the controller performs coordinated calculations based on the expected adjustment amount, progressively increasing the adjustment amount while maintaining the total gas balance within the culture chamber. When the target gas needs to be injected, it simultaneously considers whether to inject or expel an appropriate amount of air to maintain total pressure stability; when the target gas concentration needs to be reduced, it primarily controls the operation of the normally closed gas pump. Finally, the embedded controller generates two independent PWM signals. The first PWM signal is used to control the speed or duty cycle of the normally closed gas pump, thereby controlling the air input flow rate. The second pulse width modulation signal is used to control the opening duty cycle of the normally closed solenoid valve, thereby controlling the instantaneous injection flow rate of the target gas.

[0042] Through a collaborative control strategy, the system can not only accurately regulate the concentration of the target gas, but also effectively maintain the pressure stability inside the culture chamber, avoiding the shear force effect on cells or the pressure shock of the sealing system caused by sudden changes in the flow rate of a single gas.

[0043] In one embodiment of the present invention, for further explanation and limitation, the embedded controller is specifically used to determine the target control component as a temperature control component and determine the control signal of the temperature control component when the temperature deviation does not meet the corresponding target parameter deviation condition.

[0044] The temperature control components include a gas temperature control component and an ambient temperature control component. The control signal determination process for any temperature control component includes: when the temperature deviation is less than a preset threshold, the pulse width modulation signal controls the temperature control component to perform continuously adjustable heating with an equivalent power at a corresponding duty cycle, adaptively achieving temperature stability; when the temperature deviation is greater than the preset threshold, the pulse width modulation signal is in an invalid state or controls the temperature control component to stop working.

[0045] In this embodiment of the invention, a pulse width modulation (PWM) signal is generated based on the magnitude and direction of the temperature deviation, serving as the control signal for driving the temperature control component. A preset threshold can be set to 0. The temperature deviation is a signed value; a positive deviation indicates the actual temperature is higher than the target temperature, and a negative deviation indicates the actual temperature is lower than the target temperature. When the temperature deviation is less than 0, it indicates the actual temperature is lower than the target temperature, requiring heating to be initiated. At this time, the controller generates a PWM signal to control the determined temperature control component (such as a heating wire or heating pad) to perform continuously adjustable heating with an equivalent power at a corresponding duty cycle, adaptively achieving temperature stability and gently and precisely increasing the temperature. When the temperature deviation is greater than 0, it indicates the actual temperature is already higher than the target temperature, requiring further heating to be avoided. At this time, the controller will immediately disable the PWM signal or directly output a command to stop the temperature control component from working. This effectively cuts off the heat source, prevents temperature overshoot, and allows the temperature to drop back to the target range through natural system heat dissipation.

[0046] By setting a preset threshold to zero and directly comparing it with a signed deviation, the controller can clearly distinguish between two basic states: heating required and heating stopped. For the heating required state, intermittent heating using pulse-modulated signals achieves fine control; for the heating stopped state, the heating output is decisively cut off to ensure safety and accuracy.

[0047] In one embodiment of the invention, for further explanation and definition, the gas temperature control component is a water bottle heating sleeve, which wraps around the mixed gas water bottle; the ambient temperature control component is a ceramic heating plate, which is arranged in the non-observation area of ​​the ITO (Indium Tin Oxide) glass cover of the cell culture chamber. The heating plate is attached to the side of the chamber with thermally conductive adhesive. The ITO cover itself has the function of generating heat through electricity to prevent fogging from interfering with microscope observation.

[0048] In one embodiment of the invention, for further explanation and definition, the target temperature includes the target gas temperature, and the gas control component includes a mixed-gas bottle.

[0049] Specifically, the embedded control system is used to acquire the air intake volume of the mixed gas bottle and calculate the air intake volume deviation value between the air intake volume and the preset air intake volume threshold; calculate the gas temperature compensation value based on the air intake volume deviation value, and positively correct the target gas temperature through the gas temperature compensation value, so that the corrected target gas temperature is used as the target gas temperature in the subsequent calculation of the gas temperature deviation.

[0050] In this embodiment of the invention, the embedded control system possesses intelligent compensation capabilities to dynamically adjust the target gas temperature based on the actual intake volume. This avoids disturbances caused by continuous intake while ensuring the timeliness and accuracy of other adjustments in intermittent gas supply scenarios, thereby solving the problem of altered heat exchange conditions due to changes in gas flow rate, ultimately affecting the stability of the gas output temperature. The system acquires the intake volume of the mixed-gas bottle in real time through a flow sensor (such as a thermal mass flow meter). The controller compares this real-time intake volume with a preset intake volume threshold (representing the standard or reference flow rate condition of the system design) to calculate the intake volume deviation value. This deviation value also has a sign: a positive deviation indicates that the actual intake volume is higher than the preset threshold, and a negative deviation indicates that the actual intake volume is lower than the preset threshold. Based on the calculated intake volume deviation value, the embedded controller calculates the gas temperature compensation value using a preset internal compensation algorithm, such as a proportional-integral algorithm based on a thermodynamic model or a lookup table method. The sign of this compensation value is related to the intake volume deviation value. When the intake air volume increases (positive deviation), more room temperature gas enters the water bottle, leading to an increased heating load. If the original target temperature is maintained, the actual output gas temperature may decrease. Therefore, the calculated gas temperature compensation value is positive. When the intake air volume decreases (negative deviation), no compensation is needed. Therefore, when the calculated gas temperature compensation value is negative, the compensation value is set to 0. The original target gas temperature is added to the compensation value to calculate the corrected target gas temperature. This correction process is performed in real-time and in a closed loop.

[0051] It should be noted that by monitoring the intake air volume as an upstream variable and pre-compensating for the target temperature, the system can proactively offset the thermal disturbance caused by flow fluctuations, rather than passively waiting for temperature deviations to occur before making corrections.

[0052] In one embodiment of the present invention, for further explanation and limitation, the system further includes a dual MOS driving circuit; The dual MOS (Metal-Oxide-Semiconductor) drive circuit is used to receive the control signal sent by the embedded controller, amplify the control signal to obtain a drive signal, and send the drive signal to the corresponding target control component to drive the target control component to perform gas concentration and / or temperature regulation.

[0053] In this embodiment, considering that the microcontroller's driving capability in the embedded controller is insufficient to drive peripheral control components, the system also includes a dual MOS drive circuit as a reliable power interface between the embedded controller and the execution components. This circuit receives low-pressure control signals from the embedded controller for adjusting gas concentration and / or temperature, and amplifies them to generate drive signals with sufficient current and voltage driving capabilities. Subsequently, these drive signals are accurately and rapidly sent to the corresponding target control components (e.g., proportional solenoid valves, heaters, etc.), thereby directly driving these components to precisely execute the accurate proportioning of gas concentration and / or rapid and stable adjustment of temperature, ensuring high-fidelity and high-power conversion from control commands to physical actions.

[0054] In one embodiment of the present invention, for further explanation and limitation, the system further includes an interactive component; The embedded controller is further configured to output real-time temperature monitoring data of the target workstation and real-time concentration of the target gas in the cell culture chamber to the interactive component; the interactive component is configured to display the real-time temperature monitoring data and the real-time concentration of the target gas; the interactive component is further configured to acquire and forward to the embedded controller the target gas temperature, target ambient temperature, target gas concentration, and target parameter deviation conditions input by the user.

[0055] In this embodiment of the invention, to achieve human-computer interaction and intelligent monitoring, the system also includes an interactive component. Regarding information output, the embedded controller is configured to continuously collect and process key system operating parameters. It outputs real-time temperature monitoring data of the target workstation (e.g., a specific area within a cell culture chamber) and real-time concentration data of the target gas (e.g., CO2, O2, or N2) within the cell culture chamber to the interactive component. Upon receiving this data, the interactive component clearly displays the real-time temperature monitoring data and the real-time concentration of the target gas to the user through its display interface (e.g., a touchscreen or LCD screen), enabling the user to intuitively and accurately grasp the immediate status of the cell culture environment. Regarding command input, the interactive component also provides a parameter setting interface for the user. The user can input and set core control parameters such as the target gas temperature, target ambient temperature, target gas concentration, and target parameter deviation conditions (e.g., the allowable temperature fluctuation range) through this component (e.g., via a touchscreen or physical buttons). After acquiring these user inputs, the interactive component forwards them to the embedded controller. The embedded controller receives these settings and uses them as new control targets. In one application example of a microscopic in vivo culture environment control system, the system hardware architecture may include serial communication between an embedded system (embedded controller) and a capacitive touch screen (interactive component). The embedded system acquires data from a temperature-gas sensor and sends control signals to the temperature-gas control component.

[0056] This invention provides a method for controlling the in vivo culture environment for microscopes. In this embodiment, a monitoring component generates and sends real-time temperature sensing signals from the target workstation and gas concentration sensing signals from the target gas within the cell culture chamber to an embedded controller. The embedded controller determines real-time temperature monitoring data and the real-time concentration of the target gas based on the real-time temperature and gas concentration sensing signals. It calculates the temperature deviation between the real-time temperature monitoring data and the target temperature, and the gas concentration deviation between the real-time concentration and the target gas concentration. When the temperature deviation and / or gas concentration deviation do not meet the corresponding target parameter deviation conditions, a target control component and its control signal are determined based on the deviation amount that does not meet the target parameter deviation conditions. The control signal is sent to the target control component, which includes a temperature control component and / or a gas control component. The temperature control component performs temperature adjustment based on the control signal, and the gas control component performs gas concentration adjustment based on the control signal. By controlling the gas injection based on high-frequency gas detection and real-time feedback, intermittent gas supply is achieved, making the gas injection more stable and reducing gas supply vibration. High-frequency feedback temperature control also greatly reduces temperature fluctuations, thereby reducing the impact of gas fluctuations and temperature fluctuations on microscopic imaging. This reduces the interference of heat and airflow disturbances on microscopic imaging and ensures the quality of microscopic imaging.

[0057] Furthermore, as a response to the above Figure 1 The implementation of the method shown in this embodiment of the invention provides a method for controlling the in vivo culture environment for microscopes, such as... Figure 2 As shown, the method includes: 51. Through the monitoring component, real-time temperature sensing signals of the target workstation and gas concentration sensing signals of the target gas in the cell culture chamber are generated and sent to the embedded controller in real time.

[0058] 52. Using an embedded controller, based on the real-time temperature sensing signal and the gas concentration sensing signal, determine the real-time temperature monitoring data and the real-time concentration of the target gas; calculate the temperature deviation between the real-time temperature monitoring data and the target temperature, and calculate the gas concentration deviation between the real-time concentration and the target gas concentration; when the temperature deviation and / or the gas concentration deviation do not meet the corresponding target parameter deviation conditions, determine the target control component and the control signal of the target control component based on the deviation that does not meet the target parameter deviation conditions; and send the control signal to the target control component.

[0059] 53. Temperature regulation is performed by the temperature control component according to the control signal.

[0060] 54. The gas concentration is adjusted according to the control signal by means of the gas control component.

[0061] Furthermore, the real-time generation and transmission of the real-time temperature sensing signal of the target workstation and the gas concentration sensing signal of the target gas in the cell culture chamber to the embedded controller includes: Resistance data is generated and output based on temperature changes in the measurement area using a temperature sensor. The temperature sensor output resistance data is converted into analog voltage data through the temperature sensing processing circuit. The analog voltage data is converted into real-time temperature monitoring data through an analog-to-digital converter circuit. A gas concentration sensing signal is generated based on the target gas concentration within the measurement area using a non-dispersive gas sensor.

[0062] Furthermore, when the gas concentration deviation does not meet the corresponding target parameter deviation condition, the target control component is determined to be a gas control component, wherein the gas control component includes a normally closed solenoid valve for controlling the target gas input and a normally closed air pump for controlling the air input. The control signal determination process of the gas control component includes: The expected adjustment amount of the target gas is calculated based on the gas concentration deviation. Based on the expected adjustment amount, under the constraint of total gas balance in the culture chamber, a first pulse width modulation signal for controlling the normally closed gas pump and a second pulse width modulation signal for controlling the normally closed solenoid valve are generated collaboratively.

[0063] Furthermore, when the temperature deviation does not meet the corresponding target parameter deviation condition, the target control component is determined to be a temperature control component, wherein the temperature control component includes a gas temperature control component and an ambient temperature control component. The process of determining the control signal for any temperature control component includes: When the temperature deviation is less than a preset threshold, the pulse width modulation signal controls the temperature control component to perform equivalent power continuously adjustable heating with a corresponding duty cycle, thereby adaptively achieving temperature stability. When the temperature deviation exceeds a preset threshold, the pulse width modulation signal is in an invalid state or the temperature control component is stopped working.

[0064] Furthermore, the gas temperature control component is a water bottle heating sleeve, which surrounds the mixed gas water bottle; the ambient temperature control component is a ceramic heating element, which is arranged in the non-observation area of ​​the cell culture chamber.

[0065] Furthermore, the target temperature includes the target gas temperature, the gas control component includes a mixed-gas bottle, and the method further includes: Obtain the air intake volume of the mixed-gas bottle and calculate the air intake volume deviation value between the air intake volume and the preset air intake volume threshold; The gas temperature compensation value is calculated based on the intake volume deviation value. The target gas temperature is then positively corrected using the gas temperature compensation value, so that the corrected target gas temperature is used as the target gas temperature in the subsequent calculation of the gas temperature deviation.

[0066] Further, the control signal is sent to the target control component, wherein the target control component includes a temperature control component and / or a gas control component, to drive the target control component to perform gas concentration and / or temperature regulation, including: The control signal is sent to a dual MOS drive circuit to amplify the control signal and obtain a drive signal. This drive signal is then sent to the corresponding target control component to drive the target control component to perform gas concentration and / or temperature regulation.

[0067] Furthermore, the method also includes: The real-time temperature monitoring data of the target workstation and the real-time concentration of the target gas in the cell culture chamber are output to the interactive component to display the real-time temperature monitoring data and the real-time concentration of the target gas. The interactive component acquires and sends the target gas temperature, target ambient temperature, target gas concentration, and target parameter deviation conditions input by the user to the embedded controller.

[0068] This invention provides a method for controlling the in vivo culture environment for microscopes. In this embodiment, a monitoring component generates and sends real-time temperature sensing signals from the target workstation and gas concentration sensing signals from the target gas within the cell culture chamber to an embedded controller. The embedded controller determines real-time temperature monitoring data and the real-time concentration of the target gas based on the real-time temperature and gas concentration sensing signals. It calculates the temperature deviation between the real-time temperature monitoring data and the target temperature, and the gas concentration deviation between the real-time concentration and the target gas concentration. When the temperature deviation and / or gas concentration deviation do not meet the corresponding target parameter deviation conditions, a target control component and its control signal are determined based on the deviation amount that does not meet the target parameter deviation conditions. The control signal is sent to the target control component, which includes a temperature control component and / or a gas control component. The temperature control component performs temperature adjustment based on the control signal, and the gas control component performs gas concentration adjustment based on the control signal. By controlling gas injection through microsecond-level high-frequency gas detection and real-time feedback, intermittent gas supply is achieved, resulting in more stable gas injection, less gas supply vibration, and less gas consumption. High-frequency feedback temperature control effectively reduces temperature fluctuations in the imaging system, thereby reducing the impact of gas fluctuations and temperature fluctuations on microscopic imaging. This reduces the interference of heat and airflow disturbances on microscopic imaging and ensures the quality of microscopic imaging.

[0069] According to another aspect of the present invention, a workstation is provided that performs environmental parameter control operations on the cell culture chamber within the workstation based on the above-described microscopic in vivo culture environment control system.

[0070] It is obvious to those skilled in the art that the modules or steps of the present invention described above can be implemented using general-purpose computing systems. They can be centralized on a single computing system or distributed across a network of multiple computing systems. Optionally, they can be implemented using program code executable by a computing system, thereby storing them in a storage system for execution by the computing system. In some cases, the steps shown or described can be performed in a different order than those presented herein, or they can be fabricated as separate integrated circuit modules, or multiple modules or steps can be fabricated as a single integrated circuit module. Thus, the present invention is not limited to any particular combination of hardware and software.

[0071] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A microscopic in vivo culture environment control system, characterized in that, include: An embedded controller, a monitoring component, a temperature control component, and a gas control component; the embedded controller communicates with the monitoring component via an interface and controls the temperature control component and the gas control component. The monitoring component is used to generate and send real-time temperature sensing signals of the target workstation and gas concentration sensing signals of the target gas in the cell culture chamber to the embedded controller in real time. The embedded controller is configured to determine real-time temperature monitoring data and the real-time concentration of the target gas based on the real-time temperature sensing signal and the gas concentration sensing signal; calculate the temperature deviation between the real-time temperature monitoring data and the target temperature, and calculate the gas concentration deviation between the real-time concentration and the target gas concentration; when the temperature deviation and / or the gas concentration deviation do not meet the corresponding target parameter deviation conditions, determine the target control component and the control signal of the target control component based on the deviation that does not meet the target parameter deviation conditions; and send the control signal to the target control component, wherein the target control component includes a temperature control component and / or a gas control component. The temperature control component is used to perform temperature adjustment according to the control signal; The gas control component is used to perform gas concentration adjustment based on the control signal.

2. The system according to claim 1, characterized in that, The gas monitoring component includes a temperature sensor, a temperature sensing and processing circuit, an analog-to-digital conversion circuit, and a non-dispersive gas sensor. The temperature sensor is used to generate and output resistance data based on the temperature change of the measurement area; The temperature sensing processing circuit is used to convert the resistance data output by the temperature sensor into analog voltage data. The analog-to-digital converter circuit is used to convert the analog voltage data into real-time temperature monitoring data; The non-dispersive gas sensor is used to generate a gas concentration sensing signal based on the target gas concentration within the measurement area.

3. The system according to claim 1, characterized in that, The embedded controller is specifically used to determine the target control component as a gas control component and determine the control signal of the gas control component when the gas concentration deviation does not meet the corresponding target parameter deviation condition. The gas control component includes a normally closed solenoid valve for controlling the target gas input and a normally closed air pump for controlling the air input. The control signal determination process of the gas control component includes: The expected adjustment amount of the target gas is calculated based on the gas concentration deviation. Based on the expected adjustment amount, under the constraint of total gas balance in the culture chamber, a first pulse width modulation signal for controlling the normally closed gas pump and a second pulse width modulation signal for controlling the normally closed solenoid valve are generated collaboratively.

4. The system according to claim 1, characterized in that, The embedded controller is specifically used to determine the target control component as a temperature control component when the temperature deviation does not meet the corresponding target parameter deviation condition, and to determine the control signal of the temperature control component, wherein the temperature control component includes a gas temperature control component and an ambient temperature control component. The process of determining the control signal for any temperature control component includes: When the temperature deviation is less than a preset threshold, the pulse width modulation signal controls the temperature control component to perform equivalent power continuously adjustable heating with a corresponding duty cycle, thereby adaptively achieving temperature stability. When the temperature deviation exceeds a preset threshold, the pulse width modulation signal is in an invalid state or the temperature control component is stopped working.

5. The system according to claim 4, characterized in that, The gas temperature control component is a water bottle heating sleeve, which surrounds the mixed gas water bottle; the ambient temperature control component is a ceramic heating element, which is arranged in the non-observation area of ​​the cell culture chamber.

6. The system according to claim 5, characterized in that, The target temperature includes the target gas temperature, the gas control component includes a mixed gas bottle, and the embedded control system is specifically used to acquire the gas intake of the mixed gas bottle and calculate the gas intake deviation value between the gas intake and the preset gas intake threshold. The gas temperature compensation value is calculated based on the intake volume deviation value. The target gas temperature is then positively corrected using the gas temperature compensation value, so that the corrected target gas temperature is used as the target gas temperature in the subsequent calculation of the gas temperature deviation.

7. The system according to claim 1, characterized in that, The system also includes a dual MOS drive circuit; The dual MOS driving circuit is used to receive the microsecond-level high-frequency control signal sent by the embedded controller, amplify the control signal to obtain a driving signal, and send the driving signal to the corresponding target control component to drive the target control component to perform gas concentration and / or temperature regulation.

8. The system according to claim 1, characterized in that, The system also includes interactive components; The embedded controller is also used to output the real-time temperature monitoring data of the target workstation and the real-time concentration of the target gas in the cell culture chamber to the interactive component; The interactive component is used to display the real-time temperature monitoring data and the real-time concentration of the target gas; The interactive component is also used to acquire and forward to the embedded controller the target gas temperature, target ambient temperature, target gas concentration, and target parameter deviation conditions input by the user.

9. A method for controlling the environment of live culture for microscope use, characterized in that, The method is applied to a microscopic in vivo culture environment control system as described in any one of claims 1-8, comprising: The monitoring component generates and sends real-time temperature sensing signals of the target workstation and gas concentration sensing signals of the target gas in the cell culture chamber to the embedded controller in real time. The embedded controller determines real-time temperature monitoring data and the real-time concentration of the target gas based on the real-time temperature sensing signal and the gas concentration sensing signal; calculates the temperature deviation between the real-time temperature monitoring data and the target temperature, and calculates the gas concentration deviation between the real-time concentration and the target gas concentration; when the temperature deviation and / or the gas concentration deviation do not meet the corresponding target parameter deviation conditions, the controller determines the target control component and the control signal of the target control component based on the deviation that does not meet the target parameter deviation conditions; and sends the control signal to the target control component, wherein the target control component includes a temperature control component and / or a gas control component. Temperature regulation is performed by the temperature control component according to the control signal; The gas concentration is adjusted according to the control signal via the gas control component.

10. A workstation, characterized in that, The workstation performs environmental parameter control operations on the cell culture chamber within the workstation based on the microscopic in vivo culture environment control system as described in any one of claims 1-8.

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